Home

Instrument Manual

1. This is because the factory made normalization plate could be used for both single label H and 14C normalizations and dual label H C normalizations For best results 96 well plates should be used as normalization plates if 96 well sample plates are to be counted in a 1 6 detector counter The isotope 1 standard should be in G11 and the isotope 2 standard in H12 See 2 7 2 Crosstalk CPM normalization for crosstalk sample positions and 2J 18 for luminescence sample positions If background sample is selected on line 42 then thermal backgrounds for count subtraction are measured automatically An empty background cassette must be the first normalization cassette followed by the normalization cassette with standards 210 2J 23 Normalization See the table below for sample positions in normalization plates Sample position in a normalization plate po 24 well 96 well 1 6 det 1 6 det Isotope 1 standard Isotope 2 standard C single label When injection is used Standards are arranged in blocks on the normalization plate If background sample measurement is used then the background sample must be in the first position in each block on the plate see the plate maps following The plate maps for different detector configurations with 96 well plates and the maximum number of replicates are shown below If there is less than the maximum number of replicates the replicates positions are from left to right and
2. B S1 S2 S3 CY DE si cl Second plate l 2 3 4 5 6 A c4 cs e B se ss 86 C DE 86 co 45 2 Crosstalk correction 2 3 3 DPM standardization protocol Make a DPM standardization protocol by selecting in the Ready state P P rotocols then S S tandardization protocols Select edit E and a protocol number Edit the protocol see the figure below Line lee40 type 7 To exit Y Tor help I L Protocol name gt 2 Sample type 1 Normal 2 SPA 1 3 Crosstalk correction Y N N COFESCETLOR 5 Isotope 1 HS3 Zi A AO Ga Ay S35 SICE 3L 6 Paes 7 P 32 Cerenkov 8 Other 1 PMT use 1 Normal 2 Upper 3 Lower dl Window 1 5 320 10 Counting time for crosstalk s 60 0 11 Counting time for standards s a0 12 Precision 2 sigma 2 20 Number of standards 6 21 Isotope 1 activity DEM 200000 0 74 Standard curve fit selection Y N N 29 Curve fit method 1 Smoothing spline 2 Interpolation spline 3 Linear interpolation 4 Linear regression 1 Automatic smoothing Y N Y 26 Edit standard curve Y N N 30 Printer output 1 No 2 Short 3 Long 4 Programmable 3 32 Display output LINo 2 SAOLE 3 Long 4 Programmable 3 34 External output 1 No ZN SHORE 3 Long 4 Programmable 1 36 File OUEpuUt 1 No 2 SHOrE 3 Long 4 Programmable 1 40 Change special features Y N N gt gt Y Select crosstalk gt ACTIVIEY of Standards 15 given on line 21 gt Y Curve fitting metho
3. 126 2 26 Results 2 26 Results 2 26 1 Output items Different kind of output items can be entered after selecting Programmable output format 4 on lines 30 Printer output oz Display Output 34 External output 36 File output The possible output items are shown in the tables following Note in these tables y stands for 1 or 2 or 3 and z for 1 or 2 a Sample identification data AS Value of assay 1D for the current plate 0 99 POS Sample position on microtitration plate A01 H12 P24 SEO Running sample number 1 A01 96 H12 97 A01 on 2nd plate etc PR Protocol number used 0 99 CA Value of cassette ID for the current plate 0 9999 FU Value of function code for the current plate NORM STD STOP RP Repeat number of the current measurement D Detector number 1 6 S Spectrum half A B CI Cassette type 24 96 384 SH Shelf number 1 16 32 CNO Cassette order number Clilropess PLATEID Plate ID IM Injector module IMI Injector module index b Sample counting data CTIME Dead time corrected counting time in seconds ETIMEzZ Elapsed time in hours for isotope z from zero time z STIME Counting start time HH MM SS DATE Current date DD MMM YYYY TIME Current time HH MM SS SOP I Spectrum quench parameter 0 0 1024 0 SOP 1 Percentage theoretical error of spectrum quench parameter TOEP Isotope spectrum end point 0 0 1024 0 ISEP Percentage theoretica
4. NORM The cassette is to be used for CPM normalization See chapter 2 21 CPM Normalization The number of the normalization protocol is given in the PROT field STOP counting will stop after this cassette has been counted 2 4 2 2 Cassette number This is a simple number in the range 0 99 and can be output with the results If the function code is also a number then 100 cassette number function code is output in the range 0 9999 2 4 2 3 Protocol number This is normally a simple number in the range 0 99 for the exception see Stop cassette below The type of protocol it refers to depends on what is specified in the FUNC field If nothing is specified in the FUNC field then it is the number of a counting protocol 2 4 2 4 Assay protocol number This field is for MultiCalc assay protocols only Note that MultiCalc must be running when using this field otherwise the results will be ignored If counting is started from MultiCalc but there is no Assay code label then the cassette will be counted according to the other three fields but results will not be returned to MultiCalc This can be used if you want to run standardizations or normalizations You could for example have a normalization cassette counted first followed by a cassette or cassettes labelled with an assay protocol The counting protocol included in the assay protocol could make use of the normalization results just obtained See chapters 2 20 and 2 22 for more info
5. Protocols 91 to 98 inclusive are made at the factory for isotopes H Ia I E ICr 32p dual label 2H C S and P Cerenkov respectively These may not be deleted because they are connected to the corresponding counting protocols Note that no normalization is done for those protocols You must use these protocols to do the normalization before they can be used for counting unknowns Press P and N in the Ready state to choose the normalization protocol state The protocols can then be edited copied deleted printed and displayed Press F to get default parameters and E to edit a protocol C is used for copying a protocol including normalization data and D for deleting a protocol 104 Note the normalization data will be destroyed when editing an old normalization protocol The normalization must therefore be done again for this protocol Only the protocol name and password can be edited without destroying normalization data See chapter 2 25 Protocols for further details on normalization parameters and editing 2 23 4 Preparing the normalization samples Depending on which type of MicroBeta you have it is supplied from the factory with for 1 6 detector instruments with 24 or 96 well capability a 24 well normalization sample plate containing standards for H and C The H standard is in D5 and the C standard is in D6 for 12 detector instruments with 96 or 384 well capability a 96 well normalization sample plate containin
6. 2000 06 15 Then enter clock time in the format HH MM SS Where HH 1s hours in the 24 hour mode 0 23 mm minutes 0 59 and ss seconds 0 59 e g 14 05 30 It is not necessary to give seconds The default value for zero time is the current time See also chapter 2 14 Half life 44 Chemiluminescence correction Press Y to use chemiluminescence correction otherwise N default This correction is done with a correction formula see chapter 3 3 Calculation methods 45 Password Press Y to select a password 1f you want to prevent other users from changing your protocol Enter the password maximum 12 characters This will NOT be echoed to the screen You will also be asked to retype 1t to prevent typing errors The protocol cannot later be edited or deleted without knowing the password You must therefore try to remember it Only the MicroBeta supervisor has the authority to list the passwords see chapter 2 30 System Passwords He or she can help you 1f you have forgotten your password 46 Special plate Select Special plate type Wallac 1s the standard plate Betaplate filtermat plates and other 24 well plates have different dimensions and are counted using just one detector 122 2 25 Protocols However it is possible to count other 24 well plates with all detectors This selection comes after the plate dimension questions For Betaplate filtermats you can select the 96 well plate format output This is a composite of four 2
7. At any point in using MultiCalc you can get help by pressing the F10 function key To get help about the function of any softkey press the ALT key and the softkey at the same time Pressing ALT and F9 gives you a general keyboard help at any part in the program when text mode is being used for the screen To clear a help press ALT and F10 As experience in operating the instrument grows with time the need for detailed information decreases and the additional information the Help function gives will not need to be accessed so often 2 15 3 Selecting info Info contains a short user manual and information on consumables normalization and standardization positions changes after the previous program version and customizing the program Information can be viewed on the screen or printed To enter the Info state press I in the Ready State Ready gt I H elp O uit U sage C onsumables P ositions Ch a nges Cu s tomizing S y stem Info gt 73 2 15 Help and Info 2 15 4 H elp Press H to get help about the various selections 2 15 5 U sage This is a short user s manual with basics on how to use the instrument 2 15 6 C onsumables This is an up to date list of the currently available consumables 2 15 7 P ositions This is a list of sample positions in standardization and normalization 2 15 8 Ch a nges This is a list of additions and changes made to the program after the previous version 2 15 9 Cu s tom
8. European Economic Community Counting efficiency Counting efficiency for isotope 1 Counting efficiency for isotope 2 Counting efficiency window m isotope n Enhanced Graphics Adapter Variation after two recounts statistics monitor output Elapsed time in hours for isotope 1 from zero time 1 i Elapsed time in hours for isotope 2 from zero time 2 i Controls excluding of unrequired parts of the output i Exponent function Formfeed character to the printer after End of assay message Termination flag i Value of function ID code for the current plate i Function field in ID support plate Hour Too high count rate luminescence monitor output International Atomic Energy Agency International Business Machines Identifier Counting interrupted termination flag Isotope spectrum end point Theoretical percentage error for ISEP i International Standards Organization Isotope kilobyte kiloBequerel kiloelectron Volt Local area network Luminescence corrected count rate counts per second divided by 100 Theoretical percentage error for LCPS LCPS out of range monitor i Natural logarithm i Liquid scintillation counter Minute Maximum counts per channel reached termination flag MicroBeta starts program from DOS Megabyte MultiCalc Multichannel analyzer Multicolour Graphics Array 279 3 5 Abbreviations and acronyms used MEANI MEAN2 MS DOS N nCi Mean value for CCPM1 DPM1 or LCPS Mean value for C
9. The numbers which can be used as the example shows are for lt begin gt 32 datafile path 16 Protocol name number and type 8 ID label contents and shelf number 4 output column headings 2 empty line after results from one cassette l empty line after results from one row For lt rest gt the numbers are 32 end of assay 16 total count rate 8 ID label contents and shelf number 4 output column headings 2 empty line after results from one cassette l empty line after results from one row A number resulting from the addition of any combination of these numbers can only be formed by that combination So when you give the sum to the counter 1t knows what combination of these numbers has gone into making 1t and hence which lines to omit from the output Example EXCL 72 12 excludes the ID line before the first cassette lt begin gt 64 8 and excludes the ID line and headings before the other cassettes lt rest gt 8 4 Note that here 64 was added because 8 lt 12 2 26 2 Changing the format of an output field The format of an output field width of field and number of decimals can be changed The syntax is lt heading gt lt width gt 132 2 26 Results Or lt heading gt lt width gt lt decimals gt where lt heading gt One of the output items in groups a b and c listed above except S1 STAT REPL lt width gt The width of the output field in characters including lt de
10. The protocols can then be edited copied deleted printed and displayed and the curves can be plotted 65 2 12 DPM standardization Replicates of 51 and 512 are Used as calibration standards in 11 and G12 _ Twelve quench calibration standards 1toaS12 are shown in row 4 function code must be Used if start is autom atic Positions of standardization samples in a 12 detector MicroBeta Press the appropriate key for the following options F default parameters E edit a protocol C copying a protocol including standardization data D deleting a protocol Note the standardization data will be destroyed when editing an old standardization protocol The standardization must therefore be done again for this protocol Only the standardization protocol name curve fitting method the curve and password can be edited without destroying the standardization data See chapter 2 25 Protocols for further details on standardization parameters and editing 2 12 3 Preparing single label standardization samples Two to 12 DPM standards are placed in positions Al to A12 in a 96 or 384 well plate or in positions Al to B6 in a 24 well plate No empty positions in the beginning or between samples are allowed The first and last normalization standards must be also in position H11 and H12 96 well 1 6 detectors G11 and G12 96 well 12 detectors D5 and D6 24 well P23 and P24 384 well 1 6 det and M22 and M23 384 wel
11. are retrieved from a corresponding Counting protocol which is selected in the Assay protocol editor Assay protocols can only be edited in MultiCalc see Editing protocols below They are stored in both MultiCalc and in the counter New assay protocols and updates are transferred to MicroBeta by using the PARAMETERS function To get this press Fl COUNTER followed by F2 PARAMETERS The results are sent to MultiCalc in raw form for handling according to the instructions given in the MultiCalc assay protocol The great range of data handling possibilities available with MultiCalc are described in the User guide to MultiCalc functions and other manuals See especially there the chapters on Protocols Files and Quality control Assays using Assay protocols must always be started from MultiCalc Samples using Assay protocols are identified by an ASSAY label on the first cassette in a batch They can have numbers from 1 to 99 97 2 22 MultiCalc operation 2 22 5 Using MicroBeta with MultiCalc 2 22 5 1 Editing protocols MicroBeta controlled counting protocols 1 e Counting CPM normalization and DPM standardization protocols must be edited in terminal mode Select COUNTER F1 in MultiCalc s main menu press F9 one or several times to get to this level move the cursor to PLATEV3 in counter mode and select TERMINAL F3 Protocols can then be edited as described in chapter 2 25 Protocols Assay protocols which are controlled by
12. chapter 2 16 Interrupt before switching to some other application Press D again when back in UTMAC to resume counting When you want to exit completely from UTMAC choose Quit from the file menu If there is some printer output pending you will be asked whether or not you would like to print it If you have modified the setup parameters you will be asked to save them 2 31 7 10 Troubleshooting 1 Communication problems See GenTerm Troubleshooting 1 2 Disk full UTMAC periodically checks the free space left on volume s used for filing and warns you if there is too little space If any of the volumes gets really full UTMAC halts processing of data sent by counter and displays a warning message You should switch to Finder and make some space or alternatively you can close the data files Processing of counting data continues automatically when there is enough free space again or when there are no data files open Note 172 2 31 Terminal emulators however that you cannot close the temporary printer output file which is always located in the System folder of the boot volume So if you use the print through terminal option of MicroBeta make sure that you have enough free space for printer output on the boot volume 173 2 31 Terminal emulators 174 2 32 Thermostat option 2 32 Thermostat option When the Thermostat option is installed it allows the temperature of the upper detectors to be reduced and stabilized This lower
13. default and the maximum is 99 If repeat is greater than 5 results are not sorted and statistical results are not printed Repeats cannot be selected in crosstalk The number of replicates specifies how many replicates there are for a sample This is used for evaluating sample preparation errors The minimum value is 1 default and the maximum is 99 The number of cycles is the number of times a batch of sample cassettes will be counted The minimum is default and the maximum is 99 If the number of cycles is greater that then you will be asked to give the Cycle delay This is in minutes and is the time between when one cycle ends and the next one begins See chapter 2 6 Counting control for more information 42 Background correction Press Y to use background correction You can choose between using the background measured in normalization giving the position for a background sample or giving a value to be subtracted from the sample CPM Answer Yes to the background sample question 1f you want to use a background sample Enter the position for this sample H12 96 well 1 6 detectors G12 12 detectors D6 24 well or P24 384 well 1 6 detectors 1s the default Enter also the counting time 60s 1s the default Note that the background sample must be in a position reached by every detector see chapter 2 9 Detectors for these positions The measured backgrounds are subtracted from CPMs when calculating CCPMs Answer No to the
14. if necessary be separated and thus disposal costs reduced However it is essential to consult with your Radiological Safety officer and Institute Administrator before entering into any particular course of action 2 20 3 Filling wells Do not overfill wells In the case of the flexible MicroBeta 96 well plate the maximum total volume of liquid should not be much above 200 uL rigid plates have a capacity of 350 uL Splashes of scintillant on the plate should be removed with a tissue prior to applying the adhesive seal 2 20 4 Preparation of a quench series If you are using constantly quenched samples then the preparation of a quench series is not necessary However MicroBeta also offers you the possibility to handle variably quenched samples In such a case it 1s necessary to prepare a quench standard series so that the quench correction necessary to calculate the final DPM values can be made The preparation of six samples is adequate for a quench series although up to 12 samples can be used with MicroBeta Each sample should contain the same amount of radioactivity but the amount of quenching agent in the cocktail should increase incrementally so that the first sample has no quenching agent and the last the greatest amount The range of the quench standards should be such that it covers the expected quench range of the unknown samples and also that it allows the quench curve to be well defined by the curve fitting program 2 20 5 96 well
15. protocols If ParaLux is selected then AQP I is used instead of SQP I ParaLux counting has two modes High efficiency and Low background The discriminator channel that is used to calculate AQP I and High efficiency counts can also be optimized For PMT use select normal coincidence counting or use only upper or lower photomultiplier tubes Upper is the default if luminescence counting is selected Window 1 shows what the window setting is If Other has been selected as isotope 1 enter the limits for window 1 in a range from 1 to 1024 Window 3 is displayed only if Other has been selected as isotope 1 or isotope 2 in dual label counting Enter the limits for window 3 in a range from 1 to 1024 or O O if no third window is used Use mask adapter 1f you select Y for this then the mask adapter will be used for the counting Note this must be set to Y 1f you want to use the injector because the injector uses the mask adapter If you select N the parameters lines following as far as parameter 12 will not appear Use injector select Y if you actually want to use the injector system If you select N then even though the mask adapter with injectors 1s in position no injection will be done In the latter case lines from this one as far as parameter 12 will not appear Dispensing you should select automatic dispensing The other option Manual is not available at present Dispensing you can select the dispensing speed to be high
16. strip plates A1 is needed if background correction has been selected The sample array is as follows J awe 2 7 2 3 Normalization Protocol Edit a normalization protocol for crosstalk To do this in the Ready state press P P rotocols then N N ormalization protocols Select E dit and a protocol number Edit the protocol parameters see the figure overleaf for an example of the parameters 42 Protocol name Crosstalk correction Y N Isotope 1 1 H 3 ZEA AS A S35 5 Ce 5 7 P 32 Cerenkov 9 Luminescence PMT use 1 Normal 2 Upper 10 Counting time for crosstalk s ll Counting time for standards s 12 Precision 2 sigma 20 PEINE OUT pues l du RP 3 C 14 6 P 32 8 Other 3 Lower Short Programmable 34 Display OUEpILE 4 JS RO De Programmable Short Programmable Short Programmable Y N Y N 34 External output 36 File Output 1 3 Long 40 Change special features 41 Isotope activity setting 42 Background sample Y N 43 Half life correction Y N 44 Chemiluminescence correction 45 Use password Y N 46 Special plate 1 Wallac 3 Other Y N 2 4 2 4 2 4 2 4 Y N 2 BP filter 47 Strip plate 60h 60 Oe crosstalk correction X AA E E a is Z e 2 7 Crosstalk correction Select crosstalk correction on line 3 as well as the isotope line 5 and counting times lines 10 and 11 You can give the isotope activity setting on line 41 Y fo
17. switch to the new emulation later you can answer Y Example 11 system gt M Terminal emulation 1 GenTerm 2 UltroTerm 3 MultiCalc A VT52 5 VT100 6 MBW LS Number of lines on screen 24 gt Block graphics Y N Y gt Send results file to terminal Y N N gt Automatic result file deletion 14N3 Y gt Print through terminal Y N N gt Do you accept new terminal setting Y N ae Prepare your terminal for emulation change Press any key when ready Terminal type check failed Do you accept the new terminal setting Y N gt N 2J 30 11 P a sswords Press A to print protocol passwords entered on line 45 in the protocol editor see chapter 2J 25 Protocols This is possible only if no System password is in use or in Setup mode see 2 30 15 Example 12 system gt A Printing protocol passwords press to exit 2J 30 12 Customi z ing Press Z to set environment strings to customize the program The environment string name and value must be given For a list of possible strings see I nfo Customi z ing Giving D as the value sets the default value for the string Example 13 system gt Z Customized MENU N Environment string name gt MENU 241 2J 30 System MENU gt D Environment string name a 2J 30 13 MS D O S Press O to get to the MicroBeta MS DOS You must confirm this twice as example 14 shows after which the text MicroBeta A gt appears Example 14 system gt O Exit to MS DOS Y N gt Y Are
18. 3 023 stores result file 3 023 into the directory MB RESULTS of hard disk drive C 151 2 30 System d Path name B DATA gt Results are stored on the MicroBeta protocol disk B in directory DATA The character O at the head of the path shows that the MicroBeta disk 1s to be used for storing the results 2 30 10 Ter m inal Select the terminal emulation see chapter 2 31 Terminal emulators for details that your terminal or terminal PC uses The additional questions depend on the emulation selected The possible emulations are 1 GenTerm in WT emulation mode This emulation is fully tailored to be used with MicroBeta so the only things the user has to specify are see example 9 Example 9 system gt M Terminal emulation 1 GenTerm 2 UltroTerm 3 MultiCalc A VT 52 SIVIELOO 6 MBW 2 gt 1 Number of lines on screen 24 gt Print through terminal Y N N gt Do you accept the new terminal setting Y N gt N the number of lines on the screen whether the printer should be connected to the terminal PC or not see 2 30 10 5 2 UltroTerm see example 10 Select this if you are using the UltroTerm terminal emulator UltroTerm V2 0 or later is required Example 10 system gt M Terminal emulation 1 GenTerm 2 UltroTerm So Mu ie aCe LG 4 VT52 5 VT1LOO 6 MBW 2 gt Number of lines on screen 23 gt Print through terminal Y N N gt Do you accept the new terminal setting Y N gt Y Prepare your
19. 3 seconds is not monitored REC1 REC2 or ERR is printed for samples with no recounting one recounting two recountings or two recountings and still bad statistics Using the next sample function disables the statistics monitor If counting ends because of precision then the statistics monitor functions only if at least nine parts have already been counted Statistics is defined as bad if the chi square value yo O T di n i mean n 1 114 MX 0 gt 2 4 27 Here O is the observed deviation and T is the theoretical deviation The count rate during time part t is 1 and the number of parts n 10 3 3 9 Statistical calculations 3 3 9 1 Mean The mean value of corrected count rate or activity A from n repeats or replicates headings MEAN MEAN1 MEAN is calculated using the equation M 2A n 28 where the sum is over the repeat or replicate number 3 3 9 2 Theoretical standard error of mean The theoretical percentage standard error T of the mean M headings THEOR ERROR TSEM1 TSEM2 is calculated using the equation T a 100 VO SA n M 29 where 0A is the corrected count rate or activity a sigma error 261 3 3 Calculation methods 3 3 9 3 Standard error of mean The observed standard error O of the mean M in percentages headings OBS ERROR SEM1 SEM27 is calculated using the equation O a 100V gt M A n n 1 M 30 a 100V LA A n n n 1 M 3 3 9 4 Theoretical c
20. 34 External output 1 No ZORO ETE 3 Long 4 Programmable 1 gt 36 File output 1 No Z SOL E 3 Long 4 Programmable 1 gt 40 Change special features Y N N gt Y 41 Number of replicates Lo 42 Background sample Y N N gt Background counting time s 60 0 gt 43 Half life correction Y N N gt A5 Use password Y N N gt 46 Special plate 1 Wallac 2 BP filter 3 Other 1 gt 208 2J 23 Normalization One module or only the first module 1 The second of two modules 10 Both of two modules Sil The third of three modules 100 The first and third of three 101 etc up to All four of four modules ME There can be a maximum of 100 normalization protocols numbered from 0 to 99 Protocol number 0 is the default protocol Protocols 91 to 98 inclusive are made at the factory for isotopes H M S I sat Ou i Cr 322p dual label H C S and P Cerenkov respectively These may not be deleted because they are connected to the corresponding counting protocols Note that no normalization is done for those protocols You must use these protocols to do the normalization before they can be used for counting unknowns Press P and N in the Ready state to choose the normalization protocol state The protocols can then be edited copied deleted printed and displayed Press F to get default parameters and E to edit a protocol C is used for copying a protocol including normalization data and D for deleting a protocol Not
21. 41 123 125 228 229 CPM normalization 41 DPM standardization 44 Curve fit method selection stand 125 230 Customi z ing 155 241 Customizing 74 CV Mean value 262 Cycles 38 198 D Delayed start 34 Interrupt 76 Data Datafiles 49 Pause in transfer 75 310 Processing 51 Saving 50 Data bits 149 235 Data drive 150 236 Datafiles 49 Accessing 50 External 51 In MultiCalc 51 Names 50 Saving data 50 Date 27 Decay correction Half life 71 DEL key 16 188 Delay between plates 39 123 200 227 Delay time after dispensing 220 Delayed start 34 Detector assembly 266 Detector efficiency Correction 257 Normalization 255 Detector usage 1 2 or 3 Start up 292 Detectors 53 250 Number of 53 Detectors parking 300 Di isopropylnaphthalene 87 Dimensions 265 DIN 87 Discriminator channel 115 219 Disk drive Selection 49 Diskettes Disks 57 Disks Backup 58 Counter 57 Handling 57 Preparing protocol disk 59 Program 57 Program disk 8 Protocol 57 Protocol disk 8 Storing 57 Dispense 185 Dispense custom command string 245 Dispense fast command string 245 Dispense slow command string 245 Dispenser IN OUT 185 Dispensing 219 Dispensing modules 220 Dispensing offset 244 Dispensing tube ID 244 Dispensing volume 220 Dispensing volume max 245 Display output 119 224 DPM Counting 61 Standardization 63 E Counting 35 Easy DPM 64 66 125 229 Edit standard c
22. 5 Example Format has no effect format is less than length of Example Example 10 Example 3 leading spaces output resulting in a total of 10 characters ina field ie a Empty text No output mimes TO tt HH it HH HEH A method to produce 10 spaces 133 2 26 Results 2 26 4 Defining new output fields New output fields can be defined The syntax of a new output is lt heading gt lt expression gt Or lt heading gt lt expression gt lt width gt Or lt heading gt lt expression gt lt width gt lt decimals gt where lt heading gt The heading for the new output field This must not be included in the headings listed in paragraph 2 26 1 above It consists of letters and digits starting with a letter lt expression gt An arithmetic expression consisting of fixed headings listed in paragraph 2 26 1 a and b above excluding POS S STM DPMM LM FLAG W1 W2 and W3 numerical constants and operators listed below Note that statistical values cannot be used in expressions lt width gt The width of a printout field in characters default 12 lt decimals gt Number of digits to the right from the decimal point default 4 Operators allowed are Addition e g CCPM1 100 Subtraction Suda CCP ML 100 S Mudie to lie ation e g 100 CCPM1 f Division 62g CEPM1 100 4 Powerfunction Ss Oe CCPMI 0 5 SgrtE CCPM1L Abs Absolute value e g Abs CCPM1 100 Ln Natural logarithm e g
23. Acebsiation hide carck and Muth alc and Bel ulica operall on Dn operation Option 1 Printer connected to MiroBala lor outed gi seminal oper son 1 Switch on the printer 1a or 1b depending on your configuration see the appropriate figure above 2 Switch on the PC and start up the terminal emulator program or start up MultiCalc and go to the terminal option by pressing Fl COUNTER followed by F3 TERMINAL See chapters 2 31 and 2 22 respectively If you are using the workstation start the program running as described in the separate User manual 3 Insert the MicroBeta program disk into disk drive A the lower disk drive of the counter and the protocol disk into disk drive B the upper disk drive 4 Switch on MicroBeta with the power switch at the back of the counter Loading takes some 3 minutes then the counter is ready for operation Operating MicroBeta TriLux See chapters in italics for more information on any subject barcodes are fixed Cassette 1450 101 to an ID support plate which is clipped onto a cassette to show MicroBeta the counting protocol to use 2 4 A counting protocol lis a set of unction code parameters e g time isotope single dual label etc which control counting 2 24 2 6 2 7 2 12 2 14 2 19 2 33 Position Al A EET UR IL Kb TARA RA pi Count norm or stand Prot No MultiCalc Assay protocol No Counting protocols use either CPM norm
24. Betaplate filtermat plates and other 24 well plates The default dimensions 17 3 13 4 113 6 71 5 are for the Costar plate type For the Falcon plate type the dimensions 14 5 13 4 110 7 71 5 should be used The default dimensions for a special 384 well plate are 11 8 9 0 115 3 76 5 These are the distances of the centres of the Al A24 and P1 wells from the plate sides Note this line is not available if 96 well cassette has been selected elsewhere in the protocol 47 Autoquench correction Automatic quench correction can be done in single label CPM counting Isotopes must be 1 2 3 4 or 6 and a non crosstalk CPM normalization protocol must be selected 48 Delay between plates The delay between the end of plate measurement and the start of the next plate measurement can be specified The range is 0 to 9999 minutes 2J 25 8 Parameters in a normalization protocol The parameters are much the same as in the counting protocol with some exceptions Information whether normalization standardization has been done is given after the protocol name The following counting protocol parameters are not included in the normalization protocol 221 2J 25 Protocols 2 Counting mode 3 CPM normalization DPM standardization protocol 20 Count all positions in cassette 21 Cassette type Adi Counting Control The following parameters are new or changed when compared with the ones in the counting protocol 3 Crosstalk correction Press Y
25. CEPMES MEAN CVI 3 18 2 1 Beginning operation of MicroBeta TriLux Note Keyboard editing works with most of the type 2 1 3 data input 2 1 5 System operation The figure following is the flow diagram of the whole MicroBeta operating system It shows how the control letters inside parentheses lead to various functions The functions are described briefly on the following pages 19 2 1 Beginning operation of MicroBeta TriLux Ready gt H elp Dnfo C ount P rotocols S ystem Flow chart of MicroBeta TriLux commands normal level Info gt H elp Q uit U sage C onsumables P ositions Ch a nges Cu s tomizing S y stem Count gt Help Q uit nn count prot no ES Nnn norm prot no N Dnn std prot no D A utomatic counting Operate con v eyor T otal count rate Protocols gt Help Q uit C ounting protocol CPM n ormalization prot DPM s tandardization prot onveyor gt H elp Q uit O ff C lear conveyor Rack u p Rack d own Total count rate gt H elp Qhuit S how P rint R eset D efine efficiencies Counting protocol gt H elp Q uit E dit nn prot no D elete S how P rint L ist De f ault C opy PM norm protocol gt Help Qhuit E dit nn prot no D elete S how P rint L ist De f ault C opy N ormalization data M ultiCalc assay protocol i oe DPM stand protocol gt
26. FX and 12 cpi Example 7 oy Seem gt e Printer type 1 IBM 2 EpsonFX 2 gt Printer charac per inch IEO ATLA eye eee 2J 30 9 D ata drive Select diskette or hard disk drive of the terminal PC or MicroBeta on which the results are to be stored when using UltroTerm define the result file path in UltroTerm Result filing options The result file path can also be given in each protocol Example 8 system gt D Data drive LAS 2 Bs Sek eu 4 Path 4 gt Path name C MB RESULTS gt 236 2J 30 System The options are 1 diskette drive A Select this if you have a a single drive terminal PC no other possibilities or b a hard disk PC but you want to store results on the diskette e g to be transferred somewhere else 2 diskette drive B Select this if you have a dual diskette drive terminal PC Then terminal emulator software can be kept in drive A all the time e g to enable automatic restart after power failure 3 hard disk drive C Select this if your terminal PC is equipped with hard disk and you want large storage capacity If the GenTerm terminal emulator is used then results will be stored into the directory which was active when GenTerm was started 4 full disk drive and directory path Use this if options 1 3 do not match your requirements E g if you want to store results a on a hard disk drive other than C e g D b on a local area network LAN connected to the terminal PC c in a
27. If background sample is selected on line 42 then thermal backgrounds for count subtraction are measured automatically An empty background cassette must be the first normalization cassette followed by the normalization cassette with standards See the table below for sample positions in normalization plates Sample position in a normalization plate Ped 96 well 96 well 384 well 384 well PG det 1 6 det 12 det 1 6 det 12 det pf Single Dual 2 23 5 Normalization procedure First place the empty background plate in a cassette and then plate the plate with the normalization samples in a cassette which is identified with the corresponding normalization protocol number in the protocol number area and with a NORM label in the function code area Insert the cassette into the rack and close the door Press either A Automatic counting or 106 2 23 Normalization Nxx where xx 1s the normalization protocol number in the Ready or Count state to start counting If Nxx is selected the program asks Do you want to continue counting after normalization Y N gt If Y Yes 1s selected the counting continues after counting the normalization plate if N No is selected the counting stops after the normalization If normalization has already been done for the protocol then the program asks first THIS protocol contains normalization data Continue Y N gt This is to prevent accidental overwriting of normal
28. LSC solvents This makes it ideal for use with microtitration plates 2 20 2 Safety aspects The toxicological and physico chemical properties of DIN have been extensively studied see the booklet Di isopropylnaphthalene a new solvent for LSC 1989 available on request The conclusion of the study was that DIN is classified as a non hazardous substance within the meaning of the EEC Directive of the 18th September 1979 79 83 1 EEC DIN is not subject to compulsory labelling as laid down in those regulations which are effective in the European Community In accordance with UN and EEC International Transportation Regulations DIN is not required to be labelled as flammable or hazardous because its flash point exceeds 65 C it is in fact 148 C A biodegradability study of OptiPhase HiSafe II will also be found in the above mentioned booklet with the conclusion The test compound OptiPhase HiSafe II was found to be readily biodegradable by the ISO 7827 1984 E method 87 2 20 Micro volume LSC OptiPhase SuperMix belonging to the OptiPhase HiSafe family shares these same features which thus greatly simplifies the requirements involved in its transportation and disposal as well as showing it to be both safe to use and environmentally friendly In many countries it can be disposed of via the drains because of its biodegradability It is also possible to use a removable adhesive seal for the microtitration plates so that liquid and solid waste can
29. Mask Adapter Parameter 207 MAX 94 Max volumes in well 88 Maximum plate time 116 Maximum Well Volume 245 Mean value 261 Chi square test 263 Coefficient of variation 262 Standard error 262 Theoretical CV 262 Theoretical error 262 MeltiLex A 274 MicroBeta JET Background sample 211 MicroBeta JET Background sample 207 Injector system 272 Serial port 3 parameters 236 Microprocessor 252 Microtitration plate 83 Micro volume LSC 87 Mixing samples 87 Monitors 93 Out of range DPM 93 Out of range LCPS 93 Statistics 93 MS DOS 156 242 Definition 159 MultiCalc 95 Communication protocol 101 Exit to MS DOS 102 Operation 33 Printer connection 102 Start up with 291 Multichannel pump 244 Multidrop dispenser 89 N N Counting 35 Next pos 186 Next row 75 No of isotope standards stand 125 229 NORM 107 206 214 Function codes 29 Normalization 103 207 255 Absolute or relative 124 228 And MultiCalc 100 Protocol 115 218 Protocol parameters 123 227 With injection 207 Without injection 207 Normalization standardization Parameters 205 214 Number of channels 244 Number of injectors 244 Number of lines on screen 153 239 Nxx 107 206 214 O O 77 5 Index Counting 34 35 Off 75 186 Operating conditions 265 Out of range monitor DPM 93 130 LCPS 93 130 Output Display external or file 120 224 P ParaLux 61 63 67 255 ParaLux counting 115 219 Paramet
30. MultiCalc must be edited in MultiCalc Select PROTOCOLS F4 in MultiCalc s main menu and a list of Assay protocols is displayed Select CREATE F2 to create a new protocol Enter name protocol ID or number and select BETA as the technology and RIA IRMA or RATIO as assay type To edit an existing protocol move the cursor to the protocol line and press Enter If the protocol list is more than one page long the next page can be displayed by pressing the Page down key usually marked PgDn Similar the Page up key PgUp is used for scrolling one page up Press the Home key to get the first protocol and the End key to get the last protocol in the list A protocol can also be selected by giving the beginning of its name After pressing Enter the protocol with its parameters is displayed Use the cursor keys to move up and down and press Enter to change the value on a particular line The alternatives if any are displayed as softkeys F1 F8 otherwise the value is given by typing it The beginning of the protocol looks as follows 01 DUAL ASSAY 02 COUNTING TIME MAX COUNTS 03 MEASURING PARAMETERS Select dual press F1 or single F2 label results handling Note that dual assay here means the way that MultiCalc handles the results This is not the same as the counter s dual label Instead dual label counting is selected by using a dual label counting protocol Enter counting time in seconds and separated by a space the Max counts
31. MultiCalc to the drives and directories specified in the protocols These result files cannot be evaluated by MultiCalc 52 2 9 Detectors 2 9 Detectors 2 9 1 Six detector model This model of MicroBeta has twelve photomultiplier tubes arranged in six pairs Each pair works in coincidence The tubes of the pair are situated on opposite sides of the counting block one above and one below the block The detectors form a 3 x 2 array see the figure below Six samples are counted simultaneously which reduces counting time to about one sixth of that for a single detector instrument When a cassette is being counted the transportation system moves it between the detector blocks The first detector then counts position Al the second A5 A3 for 24 well plates the third A9 A5 the fourth El C1 etc see the figure below At this phase only the results from position Al are output because the results must be in the order Al A2 A3 etc The cassette moves one position forward and positions A2 A6 A10 E2 E6 and E10 are counted The results from A2 are output After counting the whole row the cassette moves to the beginning of the next row e g position B1 in detector 1 The rest of the results from row A are output Ai a2 A3 Ar AS A A as 49 a10 A11 A12 B1 B B3 BA BS B B7 BS B9 B10 B11 B12 C1 C C3 Ch C5 C C7 C8 C9 C10 C11 C12 Six detector D1 D D3 D4 DS D D7 DS D9 D10 D11 D12 modeen scien E E2 E3 E4 E5 ES E7 E
32. System gt H elp Q uit MS D O S E rror beep S tatus display V ersion C lock Se t up mode R S 232C P rinter D ata drive Ter m inal P a sswords Customi z ing Detector temp 20 H elp Q uit E dit nn prot no D elete S how P rint L ist De f ault C opy R eplot ultiCalc assay protocol gt Help Q uit S how Print L ist MicroBeta A gt MB Back to Ready gt 2 1 Beginning operation of MicroBeta TriLux MicroBeta TriLux commands normal level with a brief description of each Operations in the Ready State H elp Display Help text Dnfo Get information on using the instrument Cjount Start counting or operate conveyor P rotocols Edit counting parameters S ystem Edit system parameters Count state operations are available in the Ready state too Operations in the Info State Help Display Help text Q uit Back to the Ready state U sage View a Short user s manual C onsumables View a list of consumables available P ositions View normalization and standardization positions Ch a nges View a list of changes after the previous version Cu s tomizing View information on how to customize the program S y stem View information about the system Operations in the Count State Help Display Help text Q uit Back to the Ready state nn count prot no Start automatic counting by giving protocol number Start automatic countin
33. The cassette is used when counting Skatron filtermats cut into four pieces The filtermat in a sample bag is placed between the two plates of the cassette 3 4 10 8 24 pos cassette 1450 110 This is an open based cassette The cassette is used when counting 24 well culture plates e g Costar plates or equivalent 3 4 10 9 24 pos cassette 1450 116 This 1s a two piece cassette with 24 sample holes This cassette 1s used when counting filtermats The filtermat in a sample bag is placed between the base plate and the cover plate 3 4 10 10 24 pos cassette 1450 117 This 1s a two piece cassette with 24 sample holes This cassette is used when counting 4 mL sample vials The vials are placed between the base plate and the cover plate 267 3 4 Specifications 3 4 10 11 96 pos cassette 1450 118 This is a two piece cassette with 96 sample holes This cassette is used when counting e g P labelled dot blot samples from filtermats such as 1450 423 Nylon membrane No scintillator needs to be added to the filtermat allowing reprobing assays The filtermat in a sample bag is placed between the base plate and the cover plate having solid scintillator in sample positions 3 4 10 12 384 pos cassette 1450 130 This has 384 sample holes the cassette is used when counting 384 well sample plates All the cassettes are made of white plastic with an optical surface in the sample holes 3 4 11 Sample identification system Each cassette c
34. and the counter is ready for operation 4 1 6 3 Troubleshooting If there seems to be communication problems e g characters are displayed but the keyboard does not function it is probably one of following errors Wrong terminal emulator Example The counter is configured for GenTerm WT emulation but the PC uses UltroTerm Solution When starting the program the counter will usually check the emulator in the PC and change its own settings according to this If this does not work then proceed as follows Restart the counter by putting the power switch off and on again When in the Ready state press the Esc key F3 in UltroTerm one or several times The counter should respond Please check terminal type in S ystem Ter m inal After this the keyboard will work and you can change the terminal emulation in System Terminal press S followed by M See chapter 2 31 Terminal emulators for further explanations Error in communication parameters Example The RS 232C communication parameters in the counter differ from those in the PC Solution Set the communication parameters to be the same in the counter and PC If you are changing the parameters in the counter then restart it as shown above Go to System RS 232C press S followed by R and set the parameters as described in section 2J 30 System RS 232C See section 2 31 Terminal emulators GenTerm Changing parameters in GenTerm or 2 31 Terminal emulators UltroTerm Changing parameters in UltroTerm
35. and the third to a printer or in the case of MicroBeta JET to the injector modules Should any fault occur the built in service test routines allow the service engineer to quickly diagnose the cause of the fault and due to the flexible arrangement of the electronics easily change any components where necessary 232 3 2 Routine maintenance 3 2 Routine maintenance 3 2 1 Cleaning the instrument The cassette rack should be kept clean The cassettes should be kept clean in order to prevent dirt from coming between the detector blocks to minimise the background and maximise the light collection efficiency In MicroBeta JET the optical fibres of the mask adapter injector assembly must be kept clean If necessary wipe them with a moist cloth Tubing should be washed with water using the Wash command Note refer also to the reagent manufacturer s instructions to see if there is additional information on cleaning tubing 3 2 2 ID label Check the labels on the ID support plates Those in bad condition should be replaced by new ones to guarantee correct reading of the labels When fixing ID labels on the support plate ensure that the area where the label is to be fixed is clean e g that there is no perspiration from your fingers on it 3 2 3 Power supply fan Check that the cooling fan in the power supply unit is working by listening for its sound 3 2 4 Checking the instrument To check the instrument it is recommended
36. are hidden The person responsible for the MicroBeta system can make the hidden functions visible if he or she knows the password Press T to get into the setup mode The program then asks for the password Type the password and press Enter The characters you type are not displayed on the screen If you enter a wrong password the program displays the message Incorrect password Otherwise the program shows the full System menu To set a system password exit from the MicroBeta program to MS DOS as described earlier After the text MicroBeta A gt appears type PASSWORD xxxx and press Enter xxxx means the new password If you forget the password then set a new one 246 Part 3 General description 247 248 3 1 Instrument description 3 1 Instrument description 3 1 1 Introduction This section describes the hardware features of 1450 MicroBeta TriLux and MicroBeta JET 3 1 2 Combined liquid scintillation and luminescence counter With MicroBeta TriLux you can count beta or gamma emitting samples or glow luminescence samples With MicroBeta JET you can in addition count flash luminescence samples in which reagent injection 1s required You get the luxury of several powerful counting techniques combined in one counter 3 1 3 Microtitration plate based operation In MicroBeta samples are counted in a microtitration plate Before counting the instrument itself determines which type of plate is being used The plate is placed in
37. bag and place it on a 1450 104 cassette use 1450 466 coloured sealing tape between the sample bag and detector 2 18 3 Preparation of normalization samples 24 well format The procedure is the same as with a 96 position sample support but the normalization sample position is D5 for S1 J awb 2 18 4 Normalization protocol Edit the normalization protocol so that it is suitable for luminescence measurements To do this in the Ready state press P P rotocol then N N ormalization protocols Select edit E and the protocol number 80 2 18 Luminescence counting 1 Protocol name Luminescence 3 Crosstalk correction Y N N S LSotope 1 1 ila ees 279 A Beas A S30 SVCraol WO ere 7 P 32 Cerenkov 8 Other 9 Luminescence 9 PMT use 1 Normal 2 Upper 3 Lower 2 10 Counting time s 1 0 12 Precision 2 sigma dd sz SU PELAS QUEDPUES 1 N 2 ShOrE 3 Long 4 Programmable 3 324 Display Gu tCputs L N 2 Short 3 Long 4 Programmable 3 34 External output 1 N 2 Short 3 Long 4 Programmable 1 36 File output 1 N 2 SHOLE ee 4 Programmable dl 40 Change special features Y N 1 41 Isotope activity setting Y N N 42 Background sample Y N Y Background counting time s 60 0 AS Half life correction Y N N 45 Use password Y N N Edit the protocol parameters Select luminescence 9 on line 5 isotope 1 and counting time Is on line 11 A background plate is counted only if selected on line 42 2 18 5 Normali
38. be directly compared with each other As an example the half life of the p isotope is 14 3 days and if counting takes 24 hours the last samples will give about 5 lower activity than they would have done if counted at the same time as the first sample 2 14 2 Half life correction The half life correction is calculated if requested during protocol setting This correction is recommended if the half life of the isotope used is short and or the total counting time is long If you select one of the six defined isotopes in protocol setting the half life will appear on line 43 see the figure 40 Change special features Y N N gt Y 41 Isotope activity setting Y N N gt 42 Background sample Y N N gt 43 Half life correction Y N N gt Y E l Half life 1 h 1073500 00 gt Tritium was the isotope Zero time 1 set Y N N gt selected on line 5 and the Zero date 1 28 Jun 2000 gt half life is given Zero time 1 15 40 53 gt 18 30 automatically after line 43 44 Chemiluminescence correction Y N N gt 45 Use password Y N N gt 46 Special plate 1 Wallac 2 BP filter 3 Other 1 gt If you select isotope option 7 Other then you must specify the half life in hours on line 43 If you type 0 0 no half life correction will occur The next line asks if you want to set a zero time If you select Y then two more protocol lines appear to allow you to set the zero date and 71 2 14 Half life zero time This corre
39. because these routines involve every detector counting the same sample See chapters 2 12 and 2 23 for details 56 2 10 Diskettes 2 10 Diskettes 2 10 1 Diskettes used The diskettes used in the MicroBeta counter itself are 3 5 micro floppy disks with standard 1 44 MB format The program diskette has to be in drive A the lower drive and the protocol diskette in drive B the upper drive A diskette is inserted into a disk drive with the label upwards and towards you When the diskette is properly in its place a click is heard To remove the diskette press the button under the diskette slot Terminal emulator diskettes are normally 3 5 1 44 MB micro floppy disks and these are inserted into the A drive of the terminal PC Terminal emulator program disk 2 10 2 Diskette handling and storing A floppy disk should be handled carefully Avoid touching the magnetic surface and always when handling the diskette touch only the end with the label on Do not bend the diskette or write on it When labelling it the label should be written first and affixed afterwards on the diskette Do not place anything heavy on it e g books Put the diskette in a cardboard envelope when not in use Diskettes should be stored in a closed box to avoid dust getting on them The storage temperature should be between 10 30 C They should also be kept away from magnets 57 2 10 Diskettes 2 10 3 Backup of the terminal diskette To make a b
40. before the prompt text e g Ready H elp I nfo C ount P rotocols S ystem Ready gt The command is executed by pressing the letter either lower or upper case enclosed inside parentheses However commands that include a protocol number e g giving a pure protocol number in the Ready state to start counting of that protocol must be completed by pressing the Enter key Help text can be obtained by pressing H h or Exit to a higher level by pressing Q Pressing Control E the Ctrl and the E keys at the same time exits always to the Ready state 2 1 2 Confirming questions and messages Quite often MicroBeta displays a message such as Press any key when ready or to exit gt 15 2 1 Beginning operation of MicroBeta TriLux This kind of message is used to give you time to think if this is really the correct operation and to check that everything is prepared for the operation Press any key such as the space bar when ready to continue Otherwise you can cancel the operation by pressing slash Messages such as Do you accept the new terminal setting Y N gt are used to notify you that the operation under question can affect the functioning of the instrument counting results etc Press Y Yes to accept the operation or N No to cancel it 2 1 3 Input of data The rest of the user interface consists mostly of data input Data input differs somewhat from menu selection and confirming questions The default or t
41. cassette is used The filters are counted with the upper or lower tube and a 1450 104 filter cassette is used A special feature when using MicroBeta as a luminometer is the 1450 466 coloured sealing tape The saturation of PMTs is prevented and crosstalk between adjacent positions is minimized by cutting the intensity of luminescence light with the 1450 466 coloured sealing tape The luminescence units used are LCPS luminescence counts per second 1 e corrected CPS values 100 Before counting the actual samples the detector and hence the LCPS values should be normalized with a luminescence solution as similar to the actual samples as possible The normalization is done first and it is made use of when counting the actual samples 2 18 2 Preparation of normalization samples 96 and 384 well format Make a sample solution which corresponds to the samples to be analyzed i e has the same enzyme and substrate and pipette the same volume as in the samples into well membrane G1 1 96 well 1 12 det N22 384 well 1 6 det and M22 384 well 12 det An empty background plate is used for background measurement The 96 well sample array is as follows where S1 is the sample solution 79 2 18 Luminescence counting TAHIA W D gt Plate Close the sample plate with 1450 461 sealing tape or with 1450 466 coloured sealing tape and place on a 1450 105 cassette for counting Filter Enclose the filter in a 1450 432 plastic sample
42. cassette outside so that a robotic arm can change the plate In the MicroBeta program the robotic loading interface is selected as follows S ystem L evel T esting Detector shelf u sage Use shelves 1 1 2 16 3 32 gt 1 L evel N ormal counting The operation is controlled using the RS 232C port 2 external port of MicroBeta When the cassette is outside MicroBeta sends the text Ready and carriage return line feed The possible commands are Hor list commands Nxx start normalization using protocol number xx Dxx start standardization using prot number xx XX start counting using protocol number xx A start counting using IDs All Ready commands must be followed by a carriage return line feed or Counting can be interrupted with O After a plate has been measured and the cassette is outside MicroBeta sends the text Continue and carriage return line feed At this point O stops counting and any other character continues counting If plates are counted in the inverted position Al at the bottom right corner then Plate orientation Rotated should be selected in the counting protocol This selection reorders results so that outputs POS and SEQ are correct Warning The mask adapter has to be moved manually into the counting chamber before starting counting 231 2J 27 Robotic loading interface 232 2J 30 System 2J 30 System 2J 30 1 Selecting System System settings control the general way Mic
43. connect the cable from the injector system to the connector at the rear of MicroBeta JET see fig below Boot the system up and check the maintenance section of the User Manual page 16 After initializing and priming you should dispense at least once the volume to be used in the assay The dispensing volumes and alignment to the sample plate can be tested by running a counting protocol Counting protocol parameters are set for the dispensing speed and volumes for the selected modules 306 Part 5 Index 307 308 5 Index 16 188 dd Help 73 in path name 49 120 152 225 238 A Abbreviations used 277 Acronyms used 277 Active rows 116 221 Activity 259 Activity standardization 259 Adapters 275 Adjusted activities stand 126 230 AQP I 61 63 115 219 259 Aspirating tube I D 244 Aspirating tube length 244 ASQP I Quenching 63 Assay protocol number 30 Assay protocols 97 Asymmetric quench parameter 61 259 Automatic operation Counting terminal operation 33 Counting with MultiCalc 34 Normalization 33 Standardization 33 Automatic result file deletion 154 240 Autoquench correction 123 227 B Background 268 Background thermal 106 210 Background correction 121 129 226 256 Backslash 16 188 Backspace 16 188 Backup 309 5 Index Program diskettes 58 Protocol diskettes 58 Terminal diskette 58 Barcode 29 Barcode reader 250 Baud rate 148 2
44. corner of the screen during counting 2 3 2 Adjusting the clock When the instrument is installed you need to set the clock Apart from this the clock will not normally need resetting as it has a battery back up However if you do need to adjust the clock proceed as follows Press S System in the Ready mode then press C Clock to make changes to the clock Enter the date and time in the format described below First the program asks for the date Type it in using the format dd mmm yyyy where dd day of the month 1 31 mmm month the first three letters of the month in English 1 e JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC yyyy year four digit year number E g 10 MA Y 2000 When you have typed the date press Enter Note You can also give the date in the format Year month day if preferred e g 2000 5 10 Then the program asks for the time Type it in using the format hh mm ss where hh hours 0 23 24 hour clock mm minutes 0 59 ss seconds 0 59 optional E g 14 57 When you have done this then press Enter 21 2 3 Clock settings The last line asks Set clock When Y yes is pressed the clock is started To get back to the Ready state press Q quit to quit the System mode For more information on the System mode see chapter 2 30 System 2 3 3 Counting time During counting both the elapsed time from the beginning of counting and the set counting time are displayed on the scre
45. during counting Press P Pause printing After a while the computer asks 1f you want to pause in printing Answer Y Yes to the question to cause a pause in printing To resume printing later on press P again To not pause answer N No This feature 1s not included in MultiCalc mode 2 16 5 Causing a pause in data transfer A pause in data transfer to the terminal PC can be caused if required It is recommended that this feature be used before making a temporary exit from the terminal emulator program to the MS DOS of the terminal PC MicroBeta can measure approximately three cassettes belonging to the same assay after the pause in data transfer has occurred If the pause is longer than this then more results will not be saved 75 2 16 Interrupt To cause a pause in data transfer press D Disconnect and answer Y to the question asking confirmation Wait until the message Press D to resume data transfer appears on the live display then exit from the terminal emulator After returning to the terminal emulator press D to resume data transfer 76 2 17 Loading the cassette rack 2 17 Loading the cassette rack 2 17 1 Cassette rack The cassette rack is located inside the door and consists of 16 or 32 shelves numbered 1 to 16 32 from the bottom upwards The maximum capacity is therefore 1536 3072 samples when using 96 well plates and 384 768 samples when using 24 well plates and 6144 12288 samples when using 384 well pla
46. etc which control counting 2 24 2 6 2 7 2 12 2 14 2 19 2 33 Position Al A COCINAN ARANA Kb AEAN E E E pi Count norm or stand Prot No MultiCalc Assay protocol No Counting protocols use either ainn CPM normalization 2 23 or 1 Set u p IN Ors 2J 0 DPM standardization 2 12 2 Fix code labels to sample cassettes 2 4 info ere Eos correction 2 7 ormalization 3 Load cassettes into MicroBeta Close the ensures the counting efficiency cas ette rack compartment door 2 17 of each detector is the same i Standardization additionally 4 T o start cou nting 2 5 calculates the amount of Use the Windows Workstation e ran ana crosstalk between samples can Alte natively Dress be corrected for Normalization For MultiCalc F1 COUNTER then Enter an ene Oe counting a special set of samples For terminal etc Protocol number or A This will normally be done tely f ti For interrupt or power failure info see 2 16 and 2 13 a orai ail be stored in MicroBeta and called 5 T e results 2 2 2 20 2 32 are sent to into use in normal counting by selected output dewces 2 8 2 29 means of the counting protocol You can use the workstation a Terminal or PC 86 terminal a terminal emulator or 486 or Pentium MultiCalc to control MicroBeta Types The workstation is described in VT52 VT100 a separate user manual GenTerm MultiCale 2 22 2 31 can be UltroTerm used to make
47. for CPM counting or 2 for DPM counting 3 CPM normalization protocol DPM standardization protocol Type the number for the normalization in CPM counting or standardization protocol in DPM counting used Type H or to get a list of normalization standardization protocols Isotopes must be entered if no protocol is selected otherwise the counting protocol uses the same isotope s as in the norm std protocol Type 1f no protocol is needed This means that no normalization correction is done in CPM counting and no DPMs are calculated in DPM counting Lines 4 to 5 are displayed only if no normalization or standardization protocol number is selected Information about whether norm std has been done is given after line 3 4 Number of labels Type 1 for single label counting one isotope default or 2 for dual label counting two isotopes 5 Isotope 1 Select an isotope in dual labelled samples the isotope with the lower energy The alternatives are 1 H default 2 3 4C 4 35g Er 6 P 7 2P Cerenkov 8 Other 9 Luminescence You will be asked for window limits if you select Other otherwise window settings for the selected isotope are shown ParaLux counting can be selected in single label non crosstalk and non luminescence protocols If ParaLux is selected then AQP I is used instead of SQP 1 ParaLux counting has two modes High efficiency and Low background The discriminator channel that is used to calcula
48. for how to change the parameters in GenTerm or UltroTerm emulators or see the respective manual 4 1 6 4 Detector usage shelf usage In case of a 1 2 or 3 detector model of MicroBeta the detector usage must be defined in the system parameters before starting counting Go to the S ystem state and select the Devel Desting Then select Detector shelf u sage and choose one of the five detector alternatives 292 4 1 Installation instructions Check also that the shelf usage 1 16 or 32 shelves mask type ParaLux counting and Plate ID reader are set correctly Here you can also set the selected detector settings to all protocols Use this feature with care 4 1 7 Functional check 4 1 7 1 Preparing a functional check Carry out a functional check of the instrument including checking the function of the ID reader To do this proceed as follows Prepare four cassettes with labels On the first cassette use labels 1 3 and NORM in the function field Assay 1 Protocol 2 Cassette 3 and Function code NORM on the second 4 6 and STD on the third 7 9 and STOP and on the fourth 10 12 with the function field empty See chapter 2 4 for details of coding of a cassette Insert the first cassette into shelf 1 see the figure on the right the last into shelf 16 32 and the Last other two somewhere between cassette tj A 4 1 7 2 Starting the functional check The ID test is started in the Conveyor state on the te
49. is elapsed time from zero time to the end of counting T is the half life of the isotope and R is the background and detector efficiency corrected count rate Both single and dual label corrections have the same form Half life correction has no effect on the corrected count rate errors 3 3 4 Spectrum indexes 3 3 4 1 Isotope spectrum end point Isotope spectrum end point heading ISEP is a channel at the end of the spectrum 3 3 4 2 Spectrum quench parameter Spectrum quench parameter of the isotope spectrum heading SQP I is the spectrum mean pulse height and is calculated using the equation q 21i c i Xch 15 where 1 window low Max window high window 2 high window 3 high is the channel number and c i is the number of counts in channel 1 The SQP I counting window is truncated so that a 10 counts range is removed from both ends The a sigma percentage error of SQP I is given by a Sq q a 100 VXL Zo q DXe i q 16 3 3 4 3 Asymmetric quench parameter Asymmetric quench parameter heading AQP J is a measure of the isotope spectrum for an asymmetric sample and varies as the quench changes It establishes a relation between quench and efficiency 258 3 3 Calculation methods 3 3 5 Standardization 3 3 9 1 Activity standardization Standardization corrects for differences in sample quench levels and in detector responses All standard samples are counted with detector 1 Only the unquenched
50. memory but you can run MS DOS commands and other small programs in the remaining free memory To return to the original program type EXIT and press Enter 2 31 2 7 Exit from a terminal emulator program Exit from a program means that the current program is terminated totally to return to it you must start the program again 2 31 2 8 Backup It is recommended that you make a backup copy of your terminal emulator program diskette s before installation Use the MS DOS command DISKCOPY at the MS DOS prompt of the terminal PC See the MS DOS User s Manual of your terminal PC for details 2 31 2 9 Start up of the terminal PC A terminal PC can be started up using one of the following methods switch on power 1f power is on press the Ctrl Alt and Del keys simultaneously or press the reset button this button does not exist in all PC models 2 01 3 Conventions Manual references Interpret notations such as System Ter m inal Block graphics as chapter System section Ter m inal topic Block graphics 160 2 31 Terminal emulators 2 01 4 GenTerm 2 31 4 1 Product number and manual 1221 243 GenTerm terminal emulator 1221 921 GenTerm User Manual 2 31 4 2 Version V2 B or later 2 31 4 3 Installation a For details see the GenTerm User Manual three last pages b When the installation program asks Select emulation mode 1 Normal VT 52 terminal 2 Optimized Wallac Terminal WT then answer 2 WT emu
51. no parity DTR Port 2 External 9600 baud 8 data bits 2 stop bits no parity DTR Port 3 Printer 4800 baud 8 data bits 2 stop bits no parity DTR Normally these need not be changed However some situations when they must be are a Port 1 Terminal If the terminal PC is going to be used for other purposes during counting MultiCalc or UltroTerm allow this then it is recommended to lower the baud rate to 1200 baud or lower to ensure that characters will not be lost b Port 2 External computer The interface can depend very much on the external computer used 4 1 9 Parking the detectors If the instrument needs to be moved from the place where it was originally installed it is recommended that the detectors should be parked during the time when the instrument is actually being moved This can be done as follows Go to the S ystem state and select L evel and T est Press Q to return to the Ready state and then O to operate the conveyor Press period This will move the upper detector block against the lower one so that 1t cannot shift when the instrument is moved The following message will be seen on the screen Detectors are now parked Please turn the power off When the instrument is restarted the detector blocks will separate again automatically 300 4 2 Installation of the Injector System 4 2 Installation of the Injector System to MicroBeta JET Make a normal MicroBeta installation mechanical check and check
52. of efficiencies and background Switch the instrument off and attach the injector system to the instrument attach the injector modules to the base two screws at the bottom attach the support pieces for the solution bottles attach the transparent tubing of the solution bottles to the IN connectors of the pumps attach the black injector adapter tube No 1 to the out connector of pump No 1 and tube No 2 to the out connector of pump No 2 etc If there is more than one injector module the units are numbered ctor Module T Axle attached to 7 W the instrument Solution Bottles Injector assembly OUT 301 4 2 Installation of the Injector System attach the whole injector system to the left side of the MicroBeta JET by sliding the axles into holes in the support pieces attached to the instrument 302 4 2 Installation of the Injector System open the lid of the injector adapter module and check the tubing see fig below Systems with two injector modules have a second tube on the left side connector cut the tie which is attached to the arm for the injector assembly 303 4 2 Installation of the Injector System attach the injector assembly to the arm see fig below 304 4 2 Installation of the Injector System tie the tubing to the arm motor so the tubing can freely move into the instrument when injecting see fig below 6 l 305 4 2 Installation of the Injector System
53. position is reset This must be the first operation in setup to get the injector system ready 2J 0 6 P rime This operation fills the tubing with liquid so that there is no dead volume during dispensing Make sure the tubing is properly connected before this operation The input tubing should be in the reservoir from where the liquid is coming Press P to begin this operation It will take a few seconds You must give the number s of the injector module s to be primed and also the numbers of the channels to be used 2J 0 7 T ip This operation is to make sure that there are no drops hanging from the tips of the injectors It should be performed after prime has been done Press T to perform it You must give the number s of the injector module s to be tipped and also the numbers of the channels to be used 184 2J 0 Injector setup 2J 0 8 Disp e nser IN OUT This command should be used to move the mask adapter with its embedded injectors into the counter ready for dispensing to occur Press E to cause this to happen This command should be given before the Dispense command described next If the mask adapter is already in position in the counter and you give this command then the adapter will be moved out to the injector chamber 2J 0 9 D ispense Press D to actually cause injection to actually occur This should not be done until Init Prime and Tip have been performed Also the mask adapter should have been moved into place and
54. press Enter a gt POS CLIMB CCPME CCP Miss MEANN Vio You can also add a new heading afterwards Go to programmable output selection again Keep pressing Control N until the cursor is just after CTIME Ser POs Gl Eve Now type space and the text SQP 1 gt POS E TIME WSOP 1 _ Keep pressing Control N until the program beeps and then press Enter gt POS CTIME SOP 1 CCPML CCPMI MEANL CVIS Note Keyboard editing works with most of the type 2J 1 3 data input 190 2J 1 Beginning operation of MicroBeta JET 2J 1 5 System operation The figure following is the flow diagram of the whole MicroBeta operating system It shows how the control letters inside parentheses lead to various functions The functions are described briefly on the following pages 191 2J 1 Beginning operation of MicroBeta JET Flow chart of MicroBeta JET commands normal level Info gt onveyor gt H elp H elp Qhuit Qhuit U sage Oyff C onsumables C lear conveyor P ositions Rack u p Ch a nges Rack d own Cu s tomizing S y stem Total count rate gt Help Q uit Count gt S how Help P rint Q uit R eset nn count prot no D efine efficiencies Nnn norm prot no Counting protocol gt N Help Dnn std prot no Q uit D E dit A utomatic counting nn prot no Operate con v eyor D elete T otal count rate S how Ready gt Injector X L P rint Help L ist I nfo De f ault C
55. press Enter to start counting The Assay protocol to be used will be determined by the ID label If an Assay code is missing but there is a protocol code the samples will be counted using the counting protocol defined but results will not be returned to MultiCalc If counting was started from MultiCalc but the first cassette has no protocol code then the first assay will be counted using the default counting protocol prot No 0 but results will not be returned to MultiCalc Any other cassette that does not have an Assay protocol number will be counted with the assay protocol defined by the most recent cassette with an assay protocol number This means that within one assay you only need to label the first cassette A new label is only needed to start a new assay 2 5 1 4 Shelf number The program will then ask for the shelf number of the first cassette Give this number The default 1s shelf number 1 2 5 1 5 Delayed start Counting can also be started after a delay This 1s useful for e g incubation To do this press D when starting counting You must then enter the delay time in minutes and press Enter The program will display the remaining time before starting the counting Counting can be started immediately by pressing S or the operation can be cancelled by pressing Q Delay time before start m 60 gt S bare counting Opuit Time remaining 59 58 2 5 2 During counting The door of the counter should be properly closed
56. relative efficiency and background of each detector and then correct for it This is called normalization Normalization is done by measuring an optional background plate followed by one or two standard samples with defined activity or CPM in each detector Once CPMs have been measured the relative efficiencies can be calculated The efficiency of the detector giving the highest count rate is then defined to be one and the other detector efficiencies are expressed as a fraction of this value These fractions are called efficiency coefficients If isotope activity is given then absolute efficiencies are calculated by dividing the count rates by the activity In this way sample quench and detector absolute efficiencies can be corrected for When measuring a sample with a particular detector the CPM of the sample is corrected by dividing the CPM by the efficiency coefficient In dual label counting corrections are made using dual label correction formulae see 3 3 Calculation methods When normalization is done the results are stored with the normalization protocol The normalization data can be used by one or several protocols and is selected when editing the protocol see 2 25 Protocols 2 23 2 When is normalization necessary Each counting or assay protocol requires normalization data to be stored before it can calculate corrected CPMs for the samples counted A fresh normalization must be done when a new isotope is counted or when countin
57. sample This is used for evaluating sample preparation errors The minimum value is default and the maximum is 99 The number of cycles is the number of times a batch of sample cassettes will be counted The minimum is default and the maximum is 99 If the number of cycles is greater that then you will be asked to give the Cycle delay This is in minutes and is the time between when one cycle ends and the next one begins See chapter 2J 6 Counting control for more information 223 2J 25 Protocols 42 Background correction Press Y to use background correction You can choose between using the background measured in normalization giving the position for a background sample or giving a value to be subtracted from the sample CPM Answer Yes to the background sample question if you want to use a background sample Enter the position for this sample H12 96 well 1 6 detectors G12 12 detectors D6 24 well or P24 384 well 1 6 detectors 1s the default Enter also the counting time 60s is the default Note that the background sample must be in a position reached by every detector see chapter 2 9 Detectors for these positions The measured backgrounds are subtracted from CPMs when calculating CCPMs Answer No to the background sample question if you want to give a background value The value given in CPM is subtracted from the sample CPM for an isotope Two values are given one for each window if dual label counting is
58. sample in DPM The remaining parameters can be selected to be default 2 20 9 ParaLux counting 2 20 9 1 What is ParaLux counting In the case of scintillating samples e g SPA beads that are settled on the bottom of a well the scintillation light from the sample has to travel a longer distance through the sample solution in order to reach the upper photomultiplier tube The light reaching the lower PMT has to travel a shorter distance and it does not have to pass through much sample solution This 90 2 20 Micro volume LSC causes a scintillation light detection asymmetry because events may be detected in the lower tube which are not detected in the upper tube because of quenching This asymmetry is most pronounced with heavily colour quenched samples In the normal counting mode each pair of PMTs works in coincidence which means that both must detect the scintillation event for it to be counted However in the case described above there are true scintillation events from the sample which are detected by the lower PMT but which are not in coincidence with any event detected by the upper tube These events would normally be rejected by the coincidence system In ParaLux counting mode both the coincidence events and the non coincidence events in the lower tube are counted In addition a new quench parameter called AQP I Asymmetric Quench Parameter is used This parameter is derived from the difference between the coincidence count
59. standard N1 and the fully quenched standard N2 are counted with all detectors The SQP Ds or AQP I s efficiencies and weights of standard samples are calculated and stored for activity calculations The efficiency in a counting window is E r a 17 where r is background and half life corrected count rate and a is the given activity of the standard sample Standardization weight is given by w V k 5q S1 a 7 18 where k is the slope of the standard curve at q dq is from equation 16 and or is from equation 6 3 3 6 Activity 3 3 6 1 Using activity standardization Sample activity in disintegrations per minute headings DPM and DPM1 is A r E 19 where r is background corrected count rate 3 and E is counting efficiency 17 at SQP I or AQP Activity is half life corrected using equation 14 Efficiency E is obtained from the standardization efficiency curve of detector using the measured SQP I or AQP of detector 1 The measured SQP I or AQPO qG is first corrected to correspond to detector 1 q q 1 N2 qd N1 qd N2 qG N1 qa N2 q qa N2 20 259 3 3 Calculation methods where the first index stands for the detector number and the second index stands for the standard sample type If the measured SQP or AQP I is not between the standardization SQP I or AQP I limits then linear extrapolation with the two first or last points 1s used or spline with the two last points if s
60. terminal ready for emulation change Press any key when ready 3 MultiCalc VT52 Extensions to enable result file saving to PC disk when using MultiCalc 152 2 30 System 4 VT52 see example 11 Select this if you are using a terminal emulator that can provide only this emulation 5 VT100 This emulation is quite popular in commercial terminal emulation programs If your terminal emulator implements both VT52 and VT100 select VT100 because it supports a better range of MicroBeta features 1 e counting results displaying screen attributes result window scrolling etc 6 MBW MicroBeta Windows workstation VT52 If this is selected then results sorting and live data automatic sending must be specified The workstation program sets the terminal and other settings automatically Additional questions 2 30 10 1 Number of lines on screen Possible values from 15 to 25 the default value is 24 lines Some terminal emulator programs can scroll the whole screen if something is written at the lower right corner of screen e g column 80 on line 24 which mixes the status display If you encounter this problem give here a smaller value e g 23 lines You can select a smaller value also if your terminal PC cannot display 24 lines e g some portable PCs can display 16 lines 2 30 10 2 Block graphics For VT52 and VT100 default Y Yes If your terminal shows some strange characters around the status display and it seems that it c
61. that you must use MicroBeta JET with the injector module s installed Assays with a more stable luminescence chemistry can be measured with MicroBeta TriLux or MicroBeta JET without using injection These luminescence assays are designed to be completed on a solid support e g nylon membrane glass fibre or coated plate as well as in solution and can be directly quantified in a short time Due to the high signal glow luminescence reactions can be measured using single PMTs An optional mask adapter can be moved into place to reduce the signal by a factor of 10 if required The traditional microtitration plates are totally opaque black or white These plates are counted with the upper tube and a 1450 105 cassette is used The filters are counted with the upper or lower tube and a 1450 104 filter cassette is used A special feature when using MicroBeta as a glow type luminometer is the 1450 466 coloured sealing tape The saturation of PMTs is prevented and crosstalk between adjacent positions is minimized by cutting the intensity of luminescence light with the 1450 466 coloured sealing tape The luminescence units used are LCPS luminescence counts per second 1 e corrected CPS values 100 Before counting the actual samples the detector and hence the LCPS values should be normalized with a luminescence solution as similar to the actual samples as possible The normalization is done first and it is made use of when counting the actual sample
62. the relatively small sample volume there is such a low background count that no extra lead shielding is needed thus reducing the weight of the instrument To avoid ambient light affecting the photomultiplier tubes the inside of the instrument is painted black and the door has a light seal along its edge To protect the detectors the high voltage of the photomultiplier tubes is cut off when the door is open A warning message is displayed if counting is attempted while the door is not properly closed See chapter 2 16 Interrupt 3 1 7 Glow luminescence assays With MicroBeta TriLux luminescence assays designed to be completed on a solid support e g nylon membrane glass fibre or coated plate and in solution can be directly quantified Due to the high signal luminescence reactions can be measured using single PMTs 250 3 1 Instrument description Traditional microtitration plates are totally opaque black or white These plates are counted with the upper tube and a 1450 105 cassette is used The filters are counted with the upper or lower tube and a 1450 104 filter cassette is used A special feature when using MicroBeta as a luminometer is the 1450 466 coloured sealing tape The saturation of PMTs is prevented and crosstalk between adjacent positions 1s minimized by cutting the intensity of luminescence light with the 1450 466 coloured sealing tape The luminescence units used are LCPS luminescence counts per second 1 e correc
63. the way What follows here is a description of some of the features which are especially relevant to the operation of MultiCalc with MicroBeta The softkey sequences described begin with the main menu referred to above 2 22 2 Installation and startup The installation procedure is as follows a When you install MultiCalc on your PC you will be asked amongst other questions to select the Technology and the Counter Technology describes the counting process to be used In this case it is beta counting so you must select BETA Counter is the type of counter MultiCalc is to be working with You must select PLATEV3 You will be asked to load a Disk with communication drivers Drivers are instrument version specific so please use the instrument program disk at this point You can select more than one type of technology and counter at the same time Complete the rest of the MultiCalc installation as described in the MultiCalc User Manual 95 2 22 MultiCalc operation If MultiCalc is already installed in the PC but not for MicroBeta then you must go through a similar procedure to that described above but you select Setup only rather than Installation from the main installation menu b Start MultiCalc c Press Fl COUNTER to get to counter control d Move the cursor to PLATEV3 in the counter list e Press F5 INSTALL to install 1450 MicroBeta for MultiCalc In step a you were adding MicroBeta to the list of possible counters to b
64. this can be up to 4 modules and in multiple detector counters up to 2 modules 9 Delay time after dispensing Displayed only if Use injector has been selected to be Y The Delay Time defines the time to wait after dispensing before the counting starts There is a separate parameter for each module used The resolution is 0 1 seconds The default value is 0 1 second 10 Counting time The counting time is given in seconds The minimum value is 0 1 and the maximum is 999 999 9 seconds The default value is 60 0 If Use injector was selected for paramer 5 then there will be as many counting time parameters as there are dispensing modules 220 2J 25 Protocols 12 Precision 2 sigma Counting stops when the standard deviation is below the two sigma value in all detectors Precision is given in percent in a range from O to 99 9 O means that no precision limit is used The default value is 0 2 20 Count all positions in cassette Press Y Yes if you want to count the whole cassette default or N No if you want to specify the positions to be counted If No has been selected the next parameter will appear 21 Cassette type Type if you use cassettes with 96 wells default or 2 if the cassettes have 24 or 384 wells 22 Active rows If you answered No on line 20 you will have to specify the rows to be counted for the first cassette by typing the letters for these rows E g AC means that samples Al to A12 and Cl to C12 wil
65. to the speed you require if neither slow nor fast speeds are acceptable 2J 30 14 16 Wash command string This defines how the wash procedure occurs Wash is used to clean the tubing after use 2J 30 14 17 Tip init command string This ensures that there are no drops on the end of the tubing 2J 30 14 18 Prime command string This fills the tubing with liquid before operation can begin 2J 30 14 19 Suck back command string This command causes the liquid in the tubing to be returned to the reservoir 2J 30 14 20 Initialize command string This prepares the system for use and is the first operation before any other injection operation 2J 30 14 21 Stop command string The is the command to stop operation 2J 30 14 22 Dispensing volume max This is the maximum volume that may be in a well It is the sum of the sample volume and the dispensed volume It ensures that the well is not overfilled 2J 30 15 Q uit Press Q to quit from the System state 245 2J 30 System 2J 30 16 Se t up mode If a system password is used then the system mode menu is quite short see example 16 Example 16 H elp O ult MS D 0 S E rrorbeep S tatus display V ersion C lock Se t up mode Ready gt T Password gt H elp O ult MS D 0 S E rrorbeep Status display V ersion CLOCK RIS 2302 P rinter D ata drive Ter m inal P a sswords Customi z ing 1 jector module system gt All the functions that can affect routine use
66. to the terminal PC with a standard Centronics cable The printer is normally connected to port 3 on MicroBeta 2 31 5 10 Exit to MS DOS To get to the menu press the Esc key Select either Terminal DOS gateway temporary exit or Exit the program total exit press Enter and follow the instructions When the menu is on the screen press the Esc key to get back to the terminal mode During counting it is recommended that the user types D see chapter 2 16 Interrupt before temporary exit from UltroTerm to MS DOS It is then also possible to exit totally from UltroTerm and to use the terminal PC for running other programs After returning to UltroTerm press D again to allow MicroBeta to send data to UltroTerm See UltroTerm 2 User Manual Menu for details of other menu operations 2 31 5 11 Troubleshooting 1 Communication problems See GenTerm Troubleshooting 1 167 2 31 Terminal emulators 2 Printer errors UltroTerm shows an error window Solve the printer problem power off off line mode paper out etc and then press the number key to continue printing 3 Diskette and hard disk problems See the UltroTerm User Manual chapter 4 168 2 31 Terminal emulators 2 31 6 VT52 and VT100 compatible terminals and terminal emulators 2 31 6 1 Installation This depends on the product see the manuals which accompany the product For cabling see the chapter 4 1 Installation Define the terminal features in System Ter m inal se
67. top to bottom 3 and 6 detector models without background samples The rows E H are empty in the 3 detector model B si sisi si 2 2 fsz s2 ss 83 83 53 cC si si si s1 S2 s2 82 2 S3 S3 S3 S3__ D s si si Si S2 s2 S2 s2 S3 S3 s3 s3 211 2J 23 Normalization 3 and 6 detector models with background samples Rows E H are empty in the 3 detector model ee h3 J4 J5 j6 7 s J9 jio jun 12 C Sl S1 SI Sl S2 S2 S2 S2 3 3 3 3 D si si si Si S2 s2 S2 S2 S3 S3 S3 S3__ E B4 s4 s4 s4 BS s5 S5 S5 B6 s6 s6 s6 l and 2 detector models without background samples Rows E F are empty in the 1 detector model model F s2 s2 s2 s2 Go 0 212 2J 23 Normalization Plate maps for different detector configurations with 24 well plates and the maximum number of replicates are shown below If there is less than the maximum number of replicates the replicates positions are from left to right and top to bottom 3 and 6 detector models without background samples Rows C D are empty in the 3 detector model model model 213 2J 23 Normalization 2J 23 5 Normalization procedure First place the empty background plate in a cassette 1f you are using such a plate and then place the plate with the normalization samples in a cassette that is identified with the corresponding normalization protocol number in th
68. with counting time of module 1 repeat 2 199 2J 6 Counting control Module 2 Dispense with module 2 Wait the delay time of module 2 Count with counting time of module 2 repeat 1 Count with counting time of module 2 repeat 2 Wait cycle delay time Cycle 2 Plate 1 For all coded positions Module 1 Count with counting time of module 1 repeat 1 Count with counting time of module 1 repeat 2 Module 2 Count with counting time of module 2 repeat 1 Count with counting time of module 2 repeat 2 Wait plate delay time Plate 2 For all coded positions Module 1 Count with counting time of module 1 repeat 1 Count with counting time of module 1 repeat 2 Module 2 Count with counting time of module 2 repeat 1 Count with counting time of module 2 repeat 2 Note dispensing and background counting happens only at first cycle 2J 6 8 Delay between plates This parameter can be used to control the interval between when the counting of one sample plate has finished and the next is started The range is 0 to 9999 minutes 200 2J 18 Luminescence counting 2J 18 Luminescence counting 2J 18 1 Introduction Luminescence assays come in two main forms those making use of luminescence that has a short signal flash luminescence and those making use of that which emits a long stable signal glow luminescence Both can be measured with MicroBeta however the short emission assays require reagent addition mixing and temperature control This means
69. you sure Y N gt Y Type MB to start the program again MicroBeta A gt MB H elp LIMES C ount P rotocols S ystem Ready gt Type MB and press Enter to get back to the MicroBeta program Note Avoid going to MS DOS in routine work The main reasons to go there are to make a backup copy of the protocol or program diskette 2J 30 14 l njector Press I to set injector parameters for MicroBeta JET 242 Example 15 System gt l Mask adapter Y N Injector module Y N Raise detector Y N Multichannel pump Y N Number of channels Number of injectors Injector 1 Syringe volume 1 100uL 2 250uL 3 500uL ASpirating tabe LD O05 Aspirating tube length Zla Dispensing tube I D DOS Dispensing offset uL Injector2 Syringe volume 1 100uL 2 250uL 3 500uL Aspirating Tube IDs 01 DOS Aspirating tube length li Dispensing tube I D 0503 Dispensing offset uL Dispense fast command string S10L7IAx0v1000V4000c2700L20A0kR Dispense slow command string S10L7IAxO0v1000V2000c2700L20A0R Dispense custom command string S10L7 1Axv1000V3000c2700L20A0R Wash command string SLOTAxV38000A0R LLP ante Command Ser ing S10L70M200A0A24TR Prime command string SLOTAxOV3800A0R suck back command string SLOOAxIV3800A0R Initialize command string ZOR Stop Commend seeing T Dispensing volume max uL 250 Store data Y N 2J 30 14 1 Mask adapter a S VVVVV V 3 gt 3 gt Q gt Q gt Q gt
70. 1 CPM and on line 3 select the number of the luminescence normalization protocol defined above On line 42 select Y Background correction and on the next line Y Use normalization background Edit the other parameters normally Start counting in the Ready or Count state using automatic counting and a protocol ID label or start with the protocol number and press Enter When counting the samples the CPS values are shown on the status display The stored normalization factors are used for correcting the LCPS values in the printout The normalized LCPS values are shown after counting on the status display 206 2J 23 Normalization 2J 23 Normalization 2J 23 1 What is normalization MicroBeta JET has 1 2 3 or 6 detectors allowing it to count the respective number of samples simultaneously In order for the results from each detector to be equivalent irrespective of small variations in efficiency and background between detectors it is necessary to determine the relative efficiency and background of each detector and then correct for it This is called normalization Normalization without injection In the case when the injection modules are not used normalization is done by measuring an optional background plate followed by one or two standard samples with defined activity or CPM in each detector Once CPMs have been measured the relative efficiencies can be calculated The efficiency of the detector giving the highest count rate
71. 1024 The preset energy windows shown below are used for these isotopes when the isotope is chosen in the protocol When measuring dual labelled samples any of these window combinations can be used Note Before counting MicroBeta should be normalized for each isotope used Isotope Channel settings 5 360 5 530 P Cerenkov Other 5 1024 Luminescence 5__ 1024 If only upper or lower photomultiplier tubes are used then the default window is 150 1024 for non luminescence counting 179 2 34 Window settings 180 Part 2J Operating information for MicroBeta JET The information in these chapters is for MicroBeta JET only For the equivalent MicroBeta TriLux information the chapter with the same number in the preceding part 181 182 2J 0 Injector setup 2J 0 Injector setup 2J 0 1 Injector modules and channels When setting up the injectors you are sometimes required to specify the injector module number s and the channel number s An injector module is a pump with 1 2 3 or 6 syringes each feeding tubing that goes to the mask adapter which positions the tips of the tubes above the well s Each syringe and hence piece of tubing constitutes one channel A single detector instrument can have up to four modules each with one channel A multiple detector instrument can have up to two modules each with as many channels as there are detectors in the instrument 1 e 2 3 or 6 Module 1 Molde Channels 1 to 6 Ch
72. 19 ScintiStrips are counted with the support frame 1450 486 MicroBeta Starter kit MicroPlates Contains the MicroBeta sample plates 5 of each tapes and heat sealable foil 1450 487 MicroBeta Starter kit Filters Membranes and Solid Scintillators Contains the MicroBeta filters membranes MeltiLex sheets Sample bags and Waste bags 3 4 15 8 Adapters and Inserts 1450 108 Adapter For Eppendorf tubes or equivalent The adapters are placed on a 1450 102 cassette a bottom tape is used if needed 1450 109 Insert For 24 well cell culture plates The inserts are placed in the sample wells in order to reduce crosstalk between the wells 275 3 4 Specifications 276 3 5 Abbreviations and acronyms used 3 5 Abbreviations and acronyms used A D ABS ALT AQP AQP AS ASCII BDH BG BGND BGNDm BIOS Bk bps BS CA CASS CCPM CCPM CCPMI1 CCPM1 CCPM2 CCPM2 CCPM3 CCPM3 CCPS1 CCPS2 CCPS3 CGA ch CHISQ1 CHISQ2 CLM CNO Sample passed monitor tests monitor output Analogue to digital Absolute value Alternate key Asymmetric quench parameter of the isotope spectrum Theoretical percentage error of AQP Value of assay ID for the current plate American Standard Code for Information Interchange Laboratory reagent and equipment sales company Background Background Background in window m Basic Input Output System Background Bits per second Backspace key Value of cassette ID for the curren
73. 2 Routine maintenance 253 RS 232C 148 234 RS232C protocol 300 314 Safety 141 Sample plates 272 Sample type 125 229 Sample volume 220 Scintillation proximity assay 89 ScintiStrip 273 274 Send result files to terminal 153 239 Setup mode 156 246 Shelf number 34 Short output mode 135 SPA 89 SPA default curve fit method 125 230 Special plate 122 227 Specifications 265 Spectrum indexes 258 Spectrum quench index 258 Spectrum quench parameter 61 SQP I 61 Stability 270 Standard curve fit selection stand 125 230 Standard deviation Observed 262 Theoretical 262 Standardization 63 259 Standardization and MultiCalc 100 Standardization protocol 115 218 Parameters 124 229 Standby power supply 265 Start up ParaLux 293 Start up 8 290 Detector usage 292 Plate ID reader 293 Shelf usage 292 Troubleshooting 292 Statistical calculations 261 Statistical deviation values 129 Statistics 143 Restrictions 143 When available 143 Statistics monitor 93 130 261 Status 186 Status display 147 233 STD Function codes 29 Stop 75 STOP Function codes 29 Stop bits 149 235 Stop cassette Cassettes 30 Stop command string 245 Storing results 49 Strip plate 43 124 228 Suckback 185 Suck back command string 245 SuperMix 87 Syringe volume 244 System 74 147 233 System parameter setting 296 T Tapes 275 Technology 95 Terminal 152 238 Terminal emulation 159 Insta
74. 23 lines then change the Number of lines on screen to the maximum the PC can display 296 4 1 Installation instructions Notes 1 UltroTerm V2 00 or later is required 2 See the sections 2 31 Terminal emulators UltroTerm Installation and Terminal emulators GenTerm Troubleshooting 3 See also 4 8 2 and 4 8 5 4 1 8 1 3 MultiCalc a Set Terminal emulation to 3 MultiCalc b The MultiCalc installation procedure sets other terminal features Notes 1 MultiCalc V1 00 or later is required together with a 1450 communication protocol 2 See chapter 2 22 MultiCalc and chapter 2 31 Terminal emulators GenTerm Trouble shooting 3 See also 4 8 2 and 4 8 5 4 1 8 1 4 Other terminal emulator programs or terminals MicroBeta can also support other terminal emulator programs and pure terminals that use VT52 or VT100 terminal emulation If the user wants to utilize one of these then see section 2J 30 System Ter m inal for the possible settings Notes 1 See chapter 2 31 Terminal emulators VT52 and VT100 compatible terminals about using VT52 VT100 type terminals 2 See also GenTerm Troubleshooting in the same chapter 3 See also 4 8 2 and 4 8 5 297 4 1 Installation instructions 4 1 8 2 Printer connection Instrument Manual See chapter 2J 30 System Ter m inal Default The printer is connected to the terminal PC parallel printer port There are two possible ways to connect the printer 4 1 8 2 1 Printer c
75. 2J 30 10 Ter m inal Select the terminal emulation see chapter 2 31 Terminal emulators for details that your terminal or terminal PC uses The additional questions depend on the emulation selected The possible emulations are 1 GenTerm in WT emulation mode This emulation is fully tailored to be used with MicroBeta so the only things the user has to specify are see example 9 Example 9 system gt M Terminal emulation 1 GenTerm 2 UltroTerm 3 MultiCalc A TSS SIVTLOO 6 MBW 2 gt 1 Number of lines on screen 24 gt Print through terminal Y N N gt Do you accept the new terminal setting Y N gt N the number of lines on the screen whether the printer should be connected to the terminal PC or not see 2J 30 10 5 2 UltroTerm see example 10 Select this if you are using the UltroTerm terminal emulator UltroTerm V2 0 or later is required Example 10 system gt M Terminal emulation 1 GenTerm 2 UltroTerm 3 MalbtaGalc LANTO SAE LOO 6 MBW 2 gt Number of lines on screen 23 gt Print through terminal Y N N gt Do you accept the new terminal setting Y N gt Y Prepare your terminal ready for emulation change 238 2J 30 System Press any key when ready 3 MultiCalc VT52 Extensions to enable result file saving to PC disk when using MultiCalc 4 VT52 see example 11 Select this if you are using an emulator that can provide only this emulation 5 VT100 This emulation is quite popul
76. 3 system gt s Status display 1 No 2 Current positions 3 Whole plate gt 147 2 30 System 2 30 5 V ersion Show the program and MS DOS version numbers Example 4 system gt V MicroBeta program V4 4 Copyright C Wallac Oy 1997 All rights reserved MSDOS Vos COPyELGat EJ Maerosore Corporation 1987 All rights reserved 2 30 6 C lock This allows the date and time to be set MicroBeta has a calendar clock with battery back up so you do not have to set 1t each time the power has been switched off normally only when the instrument is installed Details of setting the clock are given in chapter 2 3 Clock When you set the clock the Terminal PC date and time are also set if GenTerm is used Example 5 oystem gt C Date 12 Jul 2000 gt 13 Jul 2000 Time 14 44 gt 9 30 Set clock Y N gt Y 2 30 7 R S 232C There are three RS 232C communication ports which can be used 1 for the Terminal PC 2 for an external computer 3 for the printer You can specify the communication parameters for each of these ports separately Give the number of the port and press Enter The first parameter that can be changed is the speed of transferring the data the Baud rate The possibilities are 300 600 1200 2400 4800 or 9600 The default value for the terminal and external ports is 9600 For the printout port it is 4800 If these values are not suitable then select the ones you need Next is the parity selection you
77. 3 4 10 Cassettes Cassettes are made of special grade polycarbonate plastic containing a high amount of white pigment to produce a very high degree of reflectivity about 90 while eliminating optical crosstalk between wells Measurement is made through the cassettes pat pend 266 3 4 Specifications 3 4 10 1 96 pos cassette 1450 101 This has 96 sample holes with a diameter of 7 8 mm Well separation is 9 mm The cassette is used when counting 1450 401 or equivalent 96 well sample plates 3 4 10 2 24 pos cassette 1450 102 This has 24 sample holes with a diameter of 13 2 mm Well separation is 18 mm This cassette is used when counting 1450 402 or equivalent 24 well sample plates 3 4 10 3 96 pos cassette 1450 103 This has 96 sample holes The cassette is used when counting 1450 407 plates 3 4 10 4 96 pos cassette 1450 104 This 1s a two piece cassette with 96 sample holes This cassette 1s used when counting filtermats The filtermat in a sample bag is placed between the base plate and the cover plate 3 4 10 5 96 pos cassette 1450 105 This has 96 sample holes This cassette is used when counting 1450 410 1450 405 1450 419 or equivalent sample plates 3 4 10 6 96 pos cassette 1450 106 This has 96 sample holes This cassette is used when counting e g Millipore MultiScreen Filtration plates or equivalent sample plates 3 4 10 7 24 pos cassette 1450 107 This is a two piece cassette with 24 sample holes
78. 3 4 5 6 A Ze Lo 34766 20137 291282 26680 34354 B 53926 T3803 54123 76060 55490 74528 21296 34828 27459 35104 27400 35487 D 55602 PEAS 59909 Lo 130 56 205 76007 CCPM2 1 2 gt 4 5 6 A 97465 96759 98349 96360 96441 TL B SM PIDIO 91742 80547 90815 183003 E 99128 96844 ITAA 96366 97342 DOI S2 D 92820 Sed 92343 1ID0S GLASS FOTOS Total Count Pate La LUESTOZOG L CCPM A EL LIL BO Ze CALI CeCe TESTA EL 36206 KBA End of assay 85 2 19 Microtitration plate format output 86 2 20 Micro volume LSC 2 20 Micro volume LSC 2 20 1 OptiPhase SuperMix cocktail In micro volume counting very small volumes of sample and cocktail have to be mixed Traditional cocktails are not well suited for this A cocktail specifically for micro volume counting has been produced it is called OptiPhase SuperMix Aqueous samples can be mixed with a minimum of shaking and sample loading can be greater than 1 to 1 see the table below The values are in ml per 10 ml of OptiPhase SuperMix cocktail at different temperatures 40 sucrose 0 23M ammonium sulphate 14 00 13 50 11 50 0 1M ammonium sulphate 16 00 14 50 13 00 Krebs Ringer original buffer 16 75 16 75 15 50 The solvent used di isopropylnaphthalene DIN is in all OptiPhase HiSafe cocktails including OptiPhase SuperMix DIN is not aggressive so it does not attack the majority of commonly used plastics e g polyethylene polypropylene polycarbonate polyester and PVC unlike other
79. 34 Block graphics 153 239 Blocked transportation system Loading cassettes 78 C Calculation methods 255 Capacity for samples 265 Cassette number 29 Cassette type 116 221 Cassettes 25 266 Coding 29 Stop 30 Change parameters GenTerm 162 Change special features 121 225 Changes 74 Changing parameters UltroTerm 166 Channel injector 251 Chemiluminescence correction 122 129 226 260 Chi square test Mean value 263 Chromatography 89 Chromium release 89 Cleaning 253 Clear conveyor 186 Clock 27 148 234 Coding 250 Coding cassettes 29 Coding filtermats 31 Colour quenching and SPA 89 Command 186 Communication drivers 95 Communication loss 149 235 Connections 265 Consumables 74 272 Adapters and inserts 275 Filtermats 273 Heatsealing foil 275 5 Index ID products 275 MeltiLex A 274 Miscellaneous 275 Plastic bags 274 Polystyrene Safe 274 Sample plates 272 ScintiStrip 273 274 Tapes 275 Control keys 17 188 Control k Hiding text 112 216 Controlling MicroBeta 15 187 Costar plate 123 227 Count all position in cassette 116 221 Count rate 255 Corrected 256 Counter 95 Counting 33 Control 37 121 197 225 During 34 Mode 114 218 Next row N 35 Next assay E 35 Off O 35 Starting 33 Stopping 35 Time 28 37 116 197 220 Time for background norm 124 228 Time for standards norm 124 228 CPM counting 61 Crosstalk 269 Crosstalk correction
80. 4 well plate format outputs If you select the Other option for plate type you must give the plate dimensions These are the distances of the centres of the Al A6 and D1 wells from the plate sides Special cassettes must be used for Betaplate filtermat plates and other 24 well plates The default dimensions 17 3 13 4 113 6 71 5 are for the Costar plate type For the Falcon plate type the dimensions 14 5 13 4 110 7 71 5 should be used The default dimensions for a special 384 well plate are 11 8 9 0 115 3 76 5 These are the distances of the centres of the Al A24 and P1 wells from the plate sides Note this line is not available if 96 well cassette has been selected elsewhere in the protocol 47 Autoquench correction Automatic quench correction can be done in single label CPM counting Isotopes must be 1 2 3 4 or 6 and a non crosstalk CPM normalization protocol must be selected 48 Delay between plates The delay between the end of plate measurement and the start of the next plate measurement can be specified The range is 0 to 9999 minutes 2 25 8 Parameters in a normalization protocol The parameters are much the same as in the counting protocol with some exceptions Information whether normalization standardization has been done is given after the protocol name The following counting protocol parameters are not included in the normalization protocol 2 Counting mode 3 CPM normalization DPM standardization protocol 20 Coun
81. 9 between all output columns to ease loading of the result file into commercial programs e g spreadsheets EXCL Controls excluding of unrequired parts of the output v FF Sends a form feed character to the printer after the End of assay message NOLIVE No status display Notes 1 Statistical deviation values will be output normally on the 2 sigma level However if item Sl exists in an output selection then for that output device deviation values will be printed on the 1 sigma level 11 Corrections made a Background correction if in protocol either 42 Background correction Y Yes and Use normalization background Y Yes OT Background sample Y Yes or Background subtraction 1 gt 0 0 CCPMl or DPM1 or Background subtraction 2 700 CCPMZ or DPMZ b Half life correction if in protocol 43 Half life correction Yes and Half life 1 gt 0 CCPM1 or DPM OF Half life 2 gt 0 CCPM2 or DPM2 c Chemiluminescence correction if in protocol 44 Chemiluminescence correction Yes d Detector efficiency and isotope spillover correction if normalization has been done for the protocol e Quench correction if standardization has been done f Crosstalk correction if in normalization or standardization 3 Crosstalk correction Yes 111 Monitors and flags in output see chapter 2 21 Monitors and flags 129 2 26 Results a Statistics monitor checks count rate variations in a s
82. CPM2 or DPM2 Microsoft Disk Operating System No nanoCurie NEWLINE Start a new line NL NO NOLIVE NORM OUT P30 Pl P2 PBS PC PET PLATE PLATEID PMT POS PR PREC PRINTER PROT PS 2 PVC RAM RECI REC2 REPL RESPI RESP1 RESP2 RESP2 ROM RP RS 232C S S Start a new line f Number No status live display Normalization ID label Out of range DPM monitor output Ortho phosphate Probability in percentages for CCPM1 DPM1 or LCPS Probability in percentages for CCPM2 or DPM2 Phosphate buffered saline Personal computer Polyethylene terephtalate Output results in microtitration plate format Plate identifier Photomultiplier tube Sample position on microtitration plate Protocol number used Precision reached termination flag Output same data as specified for printer Protocol listing l Protocol field in ID support plate Personal System 2 Polyvinyl chloride Random access memory Recounted once statistics monitor output Recounted twice statistics monitor output Replicate sample range Response CCPM1 or LCPS Theoretical percentage error for RESP Response CCPM2 Theoretical percentage error for RESP2 Read only memory Repeat number of the current measurement Recommended standard number 232 revision C for data communications Second Spectrum half i 280 S1 SD1 SD2 SEM1 SEM1 SEM2 SEM2 SEQ SH SPA 3 5 Abbreviations and acronyms used Selects 1 sigma le
83. CVZ SEMZS TCEVZS TSEMz SD Z SEMz LODZ TSEMZ CHISOZ PZ The data specified after this will be output only when a repeat or replicate mean value is available Selects 1 sigma level for all deviation output fields 1 Replicate sample range e g A01 A03 if A01 A02 and A03 are replicates Mean value of CCPMz DPMz or LCPS Percentage coefficient of variation for CCPMz DPMz or LCPS 1 Percentage standard error of mean for CCPMz DPMz or LCPS 1 Percentage theoretical coeff of variation for CCPMz DPMz or LCPS 1 Percentage theoretical stand error of mean for CEPMz DPMz Of LEBS AL Standard deviation For CCPMz DPMz or LCEPS 1 Standard error of me n or CCPM27 DPMz Or LCPS 1 Theoreticat standard deviation For CEPMZ DPMz or LCPS 1 Theoretical standard error of mean for CCPMz DPMz or LCPS 1 Reduced chi square value for CCPMz DPMz or LCPS Probability as a percentage for CCPMz DPMz or LCPS d Controlling selections SPECTRA NEWLINE NL SPSAVE PROT Plot spectra Printer output only Start new line Start new line shorthand Output spectra in compressed numerical format Include protocol listing in the beginning of output 128 2 26 Results PLATE Output results in microtitration plate format see the chapter Microtitration plate format output PRINTER Output same data as specified for printer iv TXT Produces Start of assay nn message TAB Produces tab character ASCII
84. D of each cassette continuously Focus and stop with ID reader on Read IDs continuously and show IDs on the screen Toggle printing of ID bitmaps in ordinary form Toggle printing of ID bitmaps in raw form Toggle printing of bad ID bitmaps in raw and ordinary form Move cassette to cassette type checkpoint Move cassette between zero point and D06 Move detector block up and down at mask change position Park detector block for shipping Move rack to the specified shelf Calibrate sensor offset steps Operations in the Total Count Rate State Help Q utt S how Print R eset D efine efficiencies Operations in the System State Help Q utt MS D O S E rror beep S tatus display V ersion C lock Se t up mode Setup mode contains also R S 232C P rinter D ata drive Ter m inal Display Help text Back to the Ready state Show the accumulated total count rate and activity Print the accumulated total count rate and activity Reset the accumulated total count rate and activity Define counting efficiencies of the isotopes Display Help text Back to the Ready state Exit to DOS Set error sound on or off Set status display on or off Show version number of program and MS DOS Set date and time Select Setup mode Set serial port parameters Set printer parameters Select drive for data saving results Set terminal parameters 23 2 1 Beginning
85. Date and place of issue 2 December 2000 TURKU FINLAND Name and signature or equivalent marking of authorized person Pekka Makinen Quality Assurance Manager SAA Lae 283 284 Part 4 Installation information 285 286 4 1 Installation instructions 4 1 Installation instructions Note Except for where noted these instructions are valid for both MicroBeta TriLux and MicroBeta JET 4 1 1 Environment Although normal clean laboratory conditions are usually quite satisfactory as an operational environment it is useful to take the following points into consideration If possible a separate room should be provided for the MicroBeta LSC as this allows the best control over the immediate environment Ventilation in the room should be adequate for all conditions of use the temperature should be reasonably constant at about 22 C relative humidity should not be excessive and direct sunlight should not be able to reach the instrument It is also important that the various isotopes are stored well away from the instrument in another room Only those radioactive samples that are actually measured should be in the laboratory at any time in order to keep the background at a low level 4 1 2 Electric power Three electrical outlets each having a protective earth should be available with if possible a separate power line for the instrument itself having an isolation switch and a fuse box If excessive fluctuations in the mains voltag
86. Error in output heading CTIMR and an arrow gt will appear Press the Control N keys As you do this you will see the letters POS CTIMR COUNTS I etc appearing After CCPM1 has been displayed the program will beep to inform that the end of the current value has been reached Keep Control B pressed down and you will see that characters at the end of output selection start disappearing Continue until the cursor blinks just after CTIMR _ shows the place of the cursor gt POS CTIMR_ If you go too far then stop pressing Control B and press Control N to make the lost characters visible again When the cursor is placed just after CTIMR as in the example press Backspace or the DEL key once and R will disappear Type the letter E gt POS SET Backspace or DEL pressed 189 2J 1 Beginning operation of MicroBeta JET gt POS CLIME Letter E typed Keep pressing Control N until there is a beep and you get gt POS CTIME COUNTSL CCPMI CCPM ls You can now continue typing new headings after ccpM1 E g gt POS CTIME COUNTS1 CCPM1 CCPM1 MEAN1 CV1 Now press Enter to complete editing If you later want to remove heading COUNTS1 go to line 31 to get the programmable output selection Keep pressing Control N until you are just past COUNTS1 gt POS CTIME COUNTS 1 Press Backspace or DEL 8 times so that couNTS1 and the preceding space disappears P PO GELME Keep pressing Control N until there is a beep and
87. FS GS H5 Ab Ab B C D Eb F G H A B7 LA D7 E FA G7 H A B7 C D7 E FA G H A B7 A D7 E FA G7 H as 49 A10 BE C8 DE EX FE G HE AS Bs CE DE EX FE GE HE AR B8 C8 DE EX FE G HE BY 9 D9 EY FI G9 H Ag BY 9 D9 EY FI Go H9 Ag BY 9 D9 EY FI G9 H 61H C18 D16 E10 FI G18 H10 414 614 C18 D10 E10 F16 614 H16 414 61H C18 D1 E10 FI 614 H16 A11 B11 C11 D11 E11 F11 G11 H11 A11 B11 C11 D11 E11 F11 11 H11 A11 B11 C11 D11 E11 F11 G11 H11 55 AT B12 C12 D12 E12 F12 G1 H12 A12 B12 C12 D12 E12 F12 12 H12 AT B12 C12 D12 E12 F12 12 H12 Three detector model cassette in first position Two detector model cassette in first position One detector model cassette in first position 2 9 Detectors 2 9 3 Selecting detector number The amount of detectors must be defined in the system parameters before starting counting Go to the System state by pressing S and select the test level by pressing L followed by T Then select detector usage by pressing U and choose one of the alternatives for the number of detectors Return to the normal level by pressing L followed by N before quitting to the Ready state by pressing Q 2 9 4 Twelve detector model A twelve detector MicroBeta has twenty four photomultiplier tubes arranged in twelve pairs Each
88. INSTRUMENT MANUAL june 2004 ws MicroBeta TriLux and M MicroBeta JET Liquid Scintillation and Luminescence Counters 7 gt PerkinElmer precisely Wallac 1450 MicroBeta TriLux and MicroBeta JET Liquid Scintillation and Luminescence Counters Valid for MicroBeta TriLux with software version 4 7 and MicroBeta JET with software version 5 2 p gt m PerkinElmer precisely PerkinElmer Life and Analytical Sci s Wallac Oy P O Box 10 FIN 20101 Turku Finland Tel 358 2 2678111 Fax 358 2 2678 357 Website www perkinelmer com Warning This equipment must be installed and used in accordance with the manufacturer s recommendations Installation and service must be performed by personnel properly trained and authorized by PerkinElmer Life and Analytical Sciences Failure to follow these instructions may invalidate your warranty and or impair the safe functioning of your equipment Contents Contents Part 1 Introduction Introduce e Oca srr Tre od 7 Operating MicroBeta TriLux short instructions oocccccnnccnnnnnnoonnnnnnnnnnononononnnnnnnnnnnnnnnnnnnnnnnnnnnnos 9 Operating MicroBeta JET short instructions cccoooooocncnnncnnnnnnnncnnnnnnnnonononanononnnnnononnnannnnnnnnnnns 11 Part 2 Operating information 2 1 Beginning operation of MicroBeta TriLuX ooooonnnnnnnccnnnnnnoocnnnnnnnnonononanononnnnnononnnnnnnccnnnnnns 15 Lol toco colima 29 TOC E T A 27 2 4 COGI
89. Ln CCPM1 Exp Exponent function e g Ln Exp CCPM1 CCPM1 Sqrt Square root e g Sqrt 4 0 2 0 Parenthesis e gs 1L0 CSPMI 1 00 Examples SPECIFICATION EXPLANATION CPS1 CPM1 60 Calculates Counts per second and CULDULS DLE in format 999999979999 RESP Ln Abs CCPM1 102 1087 0 78 8 2 An example of a more complicated formula The output is in the format 99999 99 134 2 26 Results Note If a new output field with the heading AQP is given in any programmable output then this output is used as the quench parameter instead of SQP I or AQP I 2 26 5 Short output mode If you select option 2 short output mode on protocol lines 30 32 34 or 36 then the program produces as output POS COPML ECBMLS CCPM2Z ESEMZS Results for each isotope will have their own line The corresponding output for DPM counting is POS DPM1 DPM1 DPM2 DPM2 For luminescence counting the output 1s POS CPS LEDS 2 26 6 Long output mode If you select option 3 long output mode on protocol lines 30 32 34 or 36 then the program produces as output PROT POS CTIME SQP I or AQP I CPM1 CCPM1 CCPM1 CCPM2 CCPM2 Results for each isotope will have their own line The corresponding output in DPM counting is PROT POS CLIMB SOPI Cor ACP TJ CPML CCPBML CCPMIS CCPMZ CCEMZS DPM1 DPM1 DPM2 DPM2 For luminescence counting the output is PRO Ss TIME POS CTIME CPSI CCPS LCRO 135 2 26 Results 2 26 De
90. MultiCalc must be in terminal mode if you want to do these operations from within it NOTE Do not move the rack up or down when a cassette 1s in the counting position Always first clear the conveyor Re start the system if the rack and transportation system becomes blocked 78 2 18 Luminescence counting 2 18 Luminescence counting See chapter 2J 18 for JET information 2 18 1 Introduction Luminescence assays come in two main forms those making use of luminescence that has a short signal and those making use of that which emits a long stable signal Both can be measured with a liquid scintillation counter however the short emission assays prove difficult due to the requirements for reagent addition mixing and temperature control More recently assays with a more stable luminescence chemistry have been described and marketed These have been targeted towards the molecular biology laboratory and have utilized both traditional luminometers and photographic film image analysis to quantify results With MicroBeta TriLux luminescence assays designed to be completed on a solid support e g nylon membrane glass fibre or coated plate and in solution can be directly quantified in a shorter time than with any of the above methods Due to the high signal luminescence reactions can be measured using single PMTs The traditional microtitration plates are totally opaque black or white These plates are counted with the upper tube and a 1450 105
91. NG cassettes ANG enana 29 2Y COMING ices E aeeacea tease 33 20 COUNTING CONTO anise assancsereactuatiaaraaRaoanarsalie ara 37 Z wl ATOSS AIE COCHON eio AT 41 2S DAE S dades 49 eT NDC UE CU OS n a E E E N T 53 DMO DOE e ici 57 Za DENCIA a A O 61 AZ DW ly Stindardi Zao ere Nee ad Ten On 63 ZNSE RT 69 De A Te E AA on Pu eaoiscinse teaiomaccdenasansctere eaten so eeciomac eee asaneceire se ectectae se aasenaeniomtey 71 E FP NC MNO heared O A AN 73 27 AOS LINUG GUE O APA no APP te taen ented aaaaeeadoadsen ae suekead ab taakiaaadeediagr ea Riatas 75 21 Loading th cassette Tacna T TF ZNO Luminescence COU sanra a a 79 2 19 Microtitrati n plate TOr Mat OU PU a a E 83 22 Micro O Md O a 87 22k Monitors AAA esas 93 2 22 NIMC ANC OPEL Ath ON spin iai linia 95 222 NOTO id osea 103 2 24 P 32 Dot DlOt quadntiticaON xs iioceoies sasdunahecant a ences 109 22 SW EOL OC ONS sd 111 DAS SMS Sas 127 AN NS A E Aa EEA 139 2 26 Salety ANG radioactive materials cana R 141 LS ASES ali 143 Dis ESO E EE 0 OPA E P no E A A E 147 2 de emita Emo Al Otis stacy atone hccat on rail he hes 159 2232 Thermostat OPIO aos 175 293 LOLA COUME sad rta elect 177 Contents Zea WING OW SeN Sa T 179 Part 2J Operating information for MicroBeta JET DAO OCO A O O O 183 211 Besimnins operation Of Micro Beta JET lt a A ii 187 JAR a AAA aie ne nadie ian a oe teaieiadena Sects Reale tat 197 ZV O Luminescence COUNTING aria ia 201 2523 INOGM VAI ZAIOU ontario 207 ZS O a See
92. PM2 or DPM2 Percentage theoretical standard error of mean for CCPM2 or DPM2 Produces Start of assay nn message i United Nations UltroTerm for Macintosh Video Graphics Array Digital Equipment Corporation video terminal type Digital Equipment Corporation video terminal type Window 1 limits Window 2 limits Window 3 limits 281 3 5 Abbreviations and acronyms used WT Wallac terminal emulation XON XOFF Flow control using XON ASCII 17 and XOFF ASCII 19 characters Al Yes Programmable output Normalization and standardization tables 282 1390 3693 i DECLARATION OF CONFORMITY FOR CE MARKING PerkinElmer We Supplier s name WALLAC OY Address PL 10 20101 TURKU FINLAND declare under our sole responsibility that the product Name type or model lot batch or serial number possibly sources and numbers of items 1450 MicroBeta TriLux Valid from serial number 4500762 1450 MicroBeta JET Valid from serial number 4500877 to which this declaration relates is in conformity with the following standard s or other normative document s Title and or number and date of issue of the standard s or other normative document s EN 50082 1 1992 EN 50081 1 1992 EN 61000 3 2 1995 A1 1998 A2 1998 A14 2000 EN 61000 3 3 1995 EN 61010 1 1993 if applicable following the provisions of the following directives Electromagnetic compatibility EMC 89 336 EEC Low voltage LV 73 23 EEC
93. ROBABILITY P1 P2 that a random sample from a correct distribution would have a larger value of Y than the observed 2 y value 263 3 3 Calculation methods 264 3 4 Specifications 3 4 Specifications 3 4 1 Physical dimensions Height 610 mm 630 mm 055mm 12det 85 kg 90 kg pO Kg 12der 3 4 2 Power Mains voltage selectable 100 115 120 220 240 V 10 50 60 Hz power consumption 360 VA Injector module 24 V 10 max 2A 3 4 3 External power supply Power failure recovery and protection can be obtained with an external UPS back up power supply provided This can be ordered from PerkinElmer Life and Analytical Sciences 3 4 4 Connections input output Serial ASCII interface RS 232 C Connector No 1 for terminal computer No 2 for mainframe or data net and No 3 for printer 3 4 5 Operating conditions Temperature 15 to 35 C Humidity max 10 80 Pollution degree 2 Installation category II Altitude 2000 m 265 3 4 Specifications 3 4 6 Sample capacity Sample capacity using 96 well sample plates is 16 32 96 1536 3072 samples using 24 well sample plates 16 32 24 384 768 samples and using 384 well sample plates 16 32 384 6144 12288 Any combination of cassettes either for 96 or 24 well sample plates or 96 or 384 well sample plates is possible 3 4 7 Sample cassette changer A z direction moving cassette rack with shelves for 16 or 32 cassettes This rack func
94. S2 CPS3 CT CTIME CTRL CV1 CV2 D DATE DCCPM1 DCCPM2 DCCPM3 DDPM1 DDPM2 DEAE DEL DET DIN DLCPS DNA DPM DPM DPM1 DPM1 DPM2 DPM2 DPMM DTR Count rate counts per second for isotope 1 i Count rate counts per second for isotope 2 i Count rate counts per second for window 3 l Cassette type i Dead time corrected counting time in seconds Control key Percentage coefficient of variation for CCPM1 DPM1 or LCPS 1 Percentage coefficient of variation for CCPM2 or DPM2 Detector number i Current date i Absolute theoretical error for CCPM1 Absolute theoretical error for CCPM2 Absolute theoretical error for CCPM3 Absolute theoretical error for DPM1 Absolute theoretical error for DPM2 Diethylaminoethyl Delete key Detector number Di isopropylnaphthalene Absolute theoretical error for LCPS Deoxyribonucleic acid Activity disintegration per minute Theoretical percentage error for DPM Activity disintegration per minute for isotope 1 Theoretical percentage error for DPM1 Activity disintegration per minute for isotope 2 i Theoretical percentage error for DPM2 DPM out of range monitor Flow control using Data Terminal Ready line 278 EEC EFF EFFI EFF2 EFFmn EGA ERR ETIME1 ETIME2 EXCL EXP FF FLAG FU FUNC h HIGH IAEA IBM ID INT ISEP ISEP ISO ISOT kB kBq keV LAN LCPS LCPS LM LN LSC m MAX MB MC MCA MCGA 3 5 Abbreviations and acronyms used
95. TY CCPM1 30699 44 1362 5 44 LAO 0 00 CCEMZ 11174909 GaGo 120 VAGO ADE 2 29 3 1 Printer output Printer output selection line 30 above demonstrates the default statistics output format In single label counting only a line for ccpm1 is generated 2 29 3 2 External output External output selection lines 34 and 35 above demonstrates a very compact statistical output format The program did not use the default statistical output format because there were statistical output fields in the programmable output i e MEAN1 SEM1 MEAN2 and SEM2 Here the user was not interested in all of the statistical values which can be selected POS GEIME ESP2ML ECCBMLS CCPM2 ELP MEAN1 SEM1 MEAN2 SEM2 A01 D LIN Zad DEZCS4 IERS A02 MED AIII ZrO VOT lea A03 TR SLIN Lids ISA let 2699 4 Oa AMAS 1 Led 2 29 3 3 File output File output selection lines 36 and 37 above demonstrates the use of the control switch STAT The output items to the left of STAT POS CTIME CCPM1 CCPM1 6 CCPM2 CCPM2 are output for each individual result The empty text definition 6 is used to shift ccpm2 and CCPM2 seven 6 1 characters to the right to leave space for the second line sem1 The output items to the right of STAT NL MEAN 12 MEAN1 TSEM1 SEM1 MEAN2 TSEM2 SEM2 are output only when the replicate or repeat mean value is available Then at the first NL a new line is output MEAN 12 outputs a commentary word MEAN under the CTIME of the last replica
96. a special cassette for counting which you can identify with barcoded labels Load the cassette into the cassette rack behind the door with the plate upwards and the ID labels towards you then close the door 3 1 4 Cassette rack and transportation system The cassette rack 1s located inside the door and consists of 16 32 shelves These are numbered to 16 32 from the bottom upwards The maximum capacity is 1536 3072 samples when using 96 well plates and 384 768 samples when using 24 well plates and 6144 12288 samples when using 384 well plates The cassette rack 1s in its middle position when being loaded A motor moves the rack up or down one step at a time Sensors are used for identifying shelf numbers and for checking if there are cassettes on the shelves and whether they are 24 96 or 384 well type A sensor will cut off the high voltage if the door 1s opened during counting in order to protect the detectors One cassette at a time is moved to the transportation system This is an X Y transfer mechanism which first moves it to the ID reader which reads the barcodes on the cassettes Then it is moved to the first counting position After a cassette has been counted it is moved back to the rack The elevator moves the cassette rack down one position and the next cassette is moved to the transportation system 249 3 1 Instrument description 3 1 5 Coding Code labels can be used to identify which protocol set of counting paramet
97. ackup copy of the terminal program or data diskette in the terminal PC use the MS DOS DISKCOPY command see 2 31 2 8 Terminal emulators Backup 2 10 4 Backup of the program diskette There should always be a backup copy of the program diskette in case something should happen to the original To make a backup copy start the MicroBeta program as normal Then type S to get to the System state and there type O for exit to MS DOS Press Y twice to confirm the exit After the prompt MicroBetaA gt appears on the screen type PROGRAM and press Enter Follow the instructions on the screen Put a new diskette into drive B so that a copy can be made Store the new copy of the program diskette in a safe place To get back to the MicroBeta program put the protocol diskette back into drive B type MB and press Enter 2 10 5 Backup of the protocol diskette Protocol modifications are saved on the protocol diskettes so it is advisable to make a backup copy of these diskettes from time to time in order to save the protocols used To make a backup copy of the protocol diskette begin in the same way as with the program diskette The procedure differs from the point when the prompt MicroBeta A gt is displayed Here type PROTOCOL BACKUP and press the Enter key Remove the program diskette and put a new protocol diskette into drive A to receive the protocols If you want to make a new empty protocol diskette then type PROTOCOL NEW instead and press the E
98. ad off the detector 1 efficiency from the quench curve made for samples counted with detector 1 see the figure overleaf 64 2 12 DPM standardization Quench correction curve made with upto twelve points Efficiency detector 1 E ficiency Corected SOP for GP 01 SGP Or detector 1 AP Y 4 The efficiency calibration curve is then used to obtain the actual detector efficiency corresponding to the detector 1 efficiency just obtained Efficiency calibration curve made with to points Efficiency detector 1 Efficiency reded Efficiency detector j efficiency 5 The measured CPM value is then divided by the actual detector efficiency to obtain the activity in DPM 2 12 2 Editing standardization parameters Standardization parameters are stored in standardization protocols see the listing in section 2 12 3 There can be a maximum of 100 standardization protocols numbered from 0 to 99 The default protocol is number 0 Protocols 91 to 95 97 to 98 are made at the factory for isotopes H e I ies e S P and P Cerenkov respectively These may not be deleted because they are connected to the corresponding counting protocols Note that no standardization has been done for those protocols 1 e there 1s no standardization data connected to the protocols Standardizations must actually be performed with the protocols to get the standardization data Press P and in the Ready state to choose the standardization protocol mode
99. ain to exit cassette coding If there are more cassettes 221 2J 25 Protocols in a batch than there are defined cassettes the rest of them are counted according to the last cassette specification W can be pressed to code additional cassettes later X can be pressed to change the cassette type 96 to 24 or 96 to 384 and vice versa for cassettes 2 to 16 see the example 20 Count all positions in cassette Y N Y gt N 21 Cassette type 1 96 2 24 3 12 i gt Ze Cassette 1 96 Rows ABCDEFGH gt R ROW AS EVEL hI 324 526 Row Bs OOLLITLOO0000 gt 56 Row Ex CO0GLLOOU000 0 Row D 000000000000 Cassette 2 96 Rows ABCDEFGH gt X Cassette 2 24 Rows ABCD gt AB Cassette 3 24 Rows ABCD gt 23 Plate orientation Select 2 if rotated plates Al at the bottom right corner are used 3 if mirrored Al at the top right corner and 4 if both rotated and mirrored Al at the bottom left corner 30 Printer output There are four printer output alternatives 1 No 2 Short 3 Long default 4 Programmable 1 No If No is selected no results are printed 2 Short If Short is selected sample results are printed using the short results format POS CCPM1 ccpM1 In dual label CCPM2 and CCPM2 are added ccpM is replaced with ppm in DPM counting In luminescence counting itis POS LCPS and LCPSS 3 Long The long output contains PROT POS CTIME SQP I Or AQP I CCPM1 CCPM1 plus ccpmM2 ccpM2 i
100. al Select here where to connect the printer Answer N No if you do not need to connect the printer to the terminal PC Connect instead the printer to serial port 3 of MicroBeta with serial cable 1221 502 Y Yes to connect the printer to the terminal PC In this case connect the printer to the parallel port of the terminal PC with a Centronics cable 1221 122 2J 30 10 6 Exit terminal setting To exit terminal setting press slash and Enter on any line Otherwise after you have answered all the questions described above the program confirms that you accept the change If not press N or else press Y which causes the following message to appear Prepare your terminal ready for emulation change Press any key when ready Now you can make the preparations possibly required for the emulation change e g to switch from GenTerm to UltroTerm When ready press any key Then if the change succeeded the program returns to the System state However if the message Terminal type check failed 240 2J 30 System appears see example 11 it means that MicroBeta has found that the terminal program does not use the same emulation as was selected MicroBeta sends an identity inquiry to the terminal and the answer sent by the terminal emulator software is different for each emulation GenTerm UltroTerm VT52 VT100 Answer N if you want to cancel the new terminal emulation However 1f you know that your terminal has the correct emulation or you want to
101. al Emulator for Macintosh 1221 926 UTMAC 1 1 User Manual 2 31 7 2 Version V 1 1 or later 2 31 7 3 Installation a for details see UTMAC User Manual chapter 3 Installation b If you want to run UTMAC automatically on starting up the Macintosh make an alias of the appropriate UTMAC setup document and put it into the Startup folder 2 31 7 4 Starting UTMAC To start UTMAC double click on the icon of the appropriate setup document or on the icon of the UTMAC application itself At the present time there is no UTMAC emulation mode in the counter but UTMAC is 100 compatible with the UltroTerm emulation mode 2 31 7 5 Changing parameters in UTMAC The main reason for communication problems is that the counter is configured to use one emulation and the terminal emulator uses some other emulation so check that the counter 1s set to UltroTerm terminal emulation mode If there still seems to be communication problems after changing to the correct emulation mode the reason can be that there are different communication parameters in the counter and the emulator Here 1s a quick guide for how to change the communication parameters in UltroTerm For further information see the UTMAC User Manual 1 Choose Communication from the Settings menu of UTMAC 2 Modify the appropriate parameters baud rate number of data bits number of stop bits parity and type of handshake 3 Press the OK button to confirm the changes 4 Save the modified p
102. al terminal port communication setting Note You can lose communication with the counter also by changing the serial port setting of your terminal alone Usually then some strange characters appear around the screen and commands do not function To get communication functioning again you can change terminal settings in the following way if you do not remember the terminal port setting of MicroBeta Try different baud rates Press e g space bar after setting each baud rate If there is no success then change the number of data bits usually 7 or 8 bits come into question and try different baud rates again Usually parity and handshake differences should not prevent normal manual terminal use Another method is to switch the counter off and on again The correct parameters terminal type baud rate and data bits are set automatically after instrument restart If they differ from the current parameters then a text like the following is shown Incorrect terminal baud rate 9600 bps used Please set terminal RS 232C parameters in S ystem R S 232C 149 2 30 System When it seems that communication is working go to the MicroBeta System mode and check the serial port settings and make sure that they are identical with settings of the terminal Example 6 system gt R RS 232C port number i Baud rate 1 300 2 600 3 1200 4 2400 5 4800 6 9600 6 gt Parity 1 None 2 Odd 3 Even 1 gt Data bits 8 gt Stop Dies 2 gt Hand
103. alization 2 23 or DPM standardization 2 12 1 Fix code labels to sample cassettes 2 4 o a oca RA 2 Load cassettes into MicroBeta Close the ensures the counting efficiency of each detector is the same cassette rack compartment door 2 77 ao 3 To start cou nting 2 5 calculates the amount of e x x chemical quenching Possible Us g the Windows Workstation crosstalk between samples can Altern atively oress be corrected for Normalization and standardization each require For MultiCalc F1 COUNTE R then Enter counting a special set of samples For terminal etc Protocol number or A a el oe cone separately from routine For interrupt Or power ilure info see 2 6 and 2 13 operation The results will be stored in MicroBeta and called 4 The results 2 21 2 26 2 32 are sent to into use in normal counting by means of the counting protocol selected output devices 2 8 2 29 You can use the workstation a Terminal or PC 86 terminal a terminal emulator or 486 or Pentium MultiCalc to control MicroBeta Types The workstation is described in VI52 VT100 a separate user manual GenTerm MultiCale 2 22 2 31 can be UltroTerm used to make assay protocols An MultiCalc assay protocol tells which Workstation counting protocol MicroBeta is to use and what should be done Poy with the data when it is obtained SS e g quality control analysis Terminal mode is available with MultiCalc and sh
104. amels 1 to 6 This figure shows the injector system for a six detector MicroBeta JET in which there are two modules each with six channels Each module and channel can be switched on or off and thus needs to be identified in parameter setting This is done by giving the number 0 or 1 in the right position to identify the modules and then 0 or 1 again in the right position to identify the channels E g Injector module number gt 1 Use channels gt 100100 would mean that there is a six detector instrument with a single injector module using channels syringes and 4 Injector module number gt 2 Use channels gt A dei de This is a six detector instrument with two modules and all six channels of the second module in use 183 2J 0 Injector setup 2J 0 2 Selecting Injector setup The MicroBeta JET injectors need setting up before they can be used This can be done by pressing command letter X when the Counting menu is shown The injector setup menu then appears as follows H elp Q uit I nit D ispense P rime LIIE W ash Suck b ack Co m mand S tatus Oc JE N ext pos G oto pos C learconveyor Disp e nser IN OUT 2J 0 3 Help Press H to get help about the various selections 2J 0 4 Q uit Press Q to quit from the Injector menu 2J 0 5 I nit Press I to initialize the injector module You must give the number s of the injector module s to be initialized Power is switched on to the module and the piston
105. ample and gives the values No statistic variation detected REC1 No variation after one recount REC2 No variation after two recounts ERR Still variation after two recounts b DPM out of range monitor selectable in DPM counting only checks quench levels in samples by comparing with the quench in standardization The following values can be displayed OK no out of range detected OUT DPM out of range c LCPS out of range monitor selectable in luminescence counting only checks counter saturation The following values can be displayed E OK no out of range detected HIGH counter saturation detected luminescence scaler is used Saturation cannot be detected d Termination flag tells if counting has been stopped The values for FLAG can be Normal termination PREC Precision reached MAX Hot sample more than 50 000 counts channel INT Interrupted by user iv Applicable on lines 32 Display output 34 External output and 36 File output Selects same output as was selected on line 30 Printer output However PRINTER does not select automatically protocol listing PROT although PROT exists in printer selection E g to select protocol listing for file output you must specify PRINTER PROT v The ExcL output item allows you to customize your output This can be useful if for example you want to prepare output for a spreadsheet program The normal output format of an assay is shown below in the table in the next colu
106. an be provided with a 1450 451 ID Support Plate for 1450 452 ID Labels An ID Support Plate has marked areas for ASSAY PROTOCOL and CASSETTE numbers 0 99 and FUNCTION codes CPM normalization DPM standardization stop This provides the information needed for Good Laboratory Practice There is also an optional Plate ID reader 3 4 12 Electronics 3 4 12 1 Environmental Electromagnetic susceptibility according to EN50082 1 Electromagnetic emissions according to EN50008 1 1 The following interconnecting cables should be used to comply with these standards Instrument to PC 1221 503 Instrument to printer 1221 502 3 4 12 2 Hardware 16 bit microprocessor controlled counting and data reduction Memory configuration 64 kbyte ROM and 1 Mbyte RAM Two 3 5 inch 1 44 Mbyte disk drives drive A for program disk and drive B for protocol disk Logarithmic A D converter energy range 2000 keV There is a dual 1024 channel multichannel analyzer for each detector Built in dead time correction 3 4 13 Performance 100 ns coincidence time 268 3 4 Specifications 3 4 13 1 Background at the factory Turku Finland With empty 1450 514 plate lt 15 CPM in wide H window lt 17 CPM in wide C window Unquenched sample with a volume of 150 microlitres high flash point cocktail unpurged in the 1450 401 96 well Sample Plate typically 6 CPM in wide H window typically 9 CPM in wide 14 C window Unquenched sample with a
107. and returns to the System state if serial port 2 or 3 setting is in question However when changing the terminal port port 1 setting the program then shows a help text that informs you to set your terminal to the same settings which were selected for MicroBeta before confirming the new setting When ready press any key The program should then ask Do you see This text Y N If the text appears it is quite sure that the communication works and you can press Y to confirm the change of terminal port setting If the text does not appear it is obvious that your new terminal and MicroBeta serial port settings differ In that case check from the printer the new MicroBeta setting and modify your terminal setting if required until you see the above text If you cannot make the communication work you have still one possibility Set the terminal to the original serial port setting Switch off the counter and switch on again Restarting MicroBeta at this phase returns the original terminal port communication setting Note You can lose communication with the counter also by changing the serial port setting of your terminal alone Usually then some strange characters appear around the screen and commands do not function To get communication functioning again you can change terminal settings in the following way if you do not remember the terminal port setting of MicroBeta Try different baud rates Press e g space bar after setting each baud rate If
108. anges to the protocol or F2 to exit without saving 2 22 5 3 Sending assay protocols to the counter Assay protocols must be stored in both MultiCalc and the counter After exiting from the editor the protocol will only be stored in MultiCalc and it must therefore be sent to the counter Go to the counter control press F9 and Fl and select PARAMETERS F2 Press F2 to send every Assay protocol or move to the protocol name and press Enter to send a particular protocol If you want to check what assay protocols have been sent to the counter you can use commands P and M in the MicroBeta Ready state This comes from the command list items P rotocols M ultiCalc assay protocol There are three functions you can select from Show displays the contents of an assay protocol Print prints out the contents of an assay protocol List prints the list of assay protocols 2 22 5 4 CPM normalizations and DPM standardizations Normalizations and standardizations are made in the same way as when using MicroBeta without MultiCalc see chapters 2 23 Normalization and 2 12 DPM standardization Go to the counter control in MultiCalc and select TERMINAL F3 Select normalization or 100 2 22 MultiCalc operation standardization protocol in the Ready state to edit the protocol Start the counting as normal from the Ready state Normalizations and standardizations can also be started from MultiCalc Check that a NORM or STD label is in the FUNC f
109. annot display block graphic characters select N No 2 30 10 3 Send result files to terminal For VT52 and VT100 The method of sending result files to the terminal PC depends on the terminal emulator software Often a terminal emulator can capture data between a start string and a stop string If your terminal emulator is capable of doing this then answer Y Yes Then MicroBeta stores a result file at first on the protocol diskette and sends it to the terminal as one block as Start OF assay ixXxX as Result data End of assay where xx is the protocol number If your terminal emulator cannot capture data then answer N No Then the result files are stored on the protocol diskette of MicroBeta but not transferred to the terminal PC 153 2 30 System Note 1 Result file will be output if protocol line 36 FILE OUTPUT is set to other than 1 No output Note 2 See chapter Results for details 2 30 10 4 Automatic result file deletion For VT52 and VT100 default Y Yes The capacity of the protocol diskette is limited 300KB 1 3MB depending on the number of protocols and it is capable of storing the results from at least 20 cassettes 1f the output selection for FILE OUTPUT does not specify too many different printout fields If this selection is set to Y Yes then the result files on protocol diskette will be deleted automatically when starting counting and after sending a result file to the terminal PC is complete If you want to ensure
110. ar in commercial terminal emulation programs If your terminal emulator implements both VT52 and VT100 select VT100 because it supports a better range of MicroBeta features 1 e counting results displaying screen attributes result window scrolling etc 6 MBW MicroBeta Windows workstation VT52 If this is selected then results sorting and live data automatic sending must be specified The workstation program sets the terminal and other settings automatically Additional questions 2J 30 10 1 Number of lines on screen Possible values from 15 to 25 the default value is 24 lines Some terminal emulator programs can scroll the whole screen if something is written at the lower right corner of screen e g column 80 on line 24 which mixes the status display If you encounter this problem give here a smaller value e g 23 lines You can select a smaller value also if your terminal PC cannot display 24 lines e g some portable PCs can display 16 lines 2J 30 10 2 Block graphics For VT52 and VT100 default Y Yes If your terminal shows some strange characters around the status display and it seems that it cannot display block graphic characters select N No 2J 30 10 3 Send result files to terminal For VT52 and VT100 The method of sending result files to the terminal PC depends on the terminal emulator software Often a terminal emulator can capture data between a start string and a stop string If your terminal emulator is capable of
111. arameters in the appropriate setup file using the Save or Save As command from the File menu Section 2 30 System RS 232C in this manual describes how to change communication parameters in the counter This section also contains the default parameter settings 171 2 31 Terminal emulators 2 31 7 6 Special features UTMAC uses an extended VT52 terminal emulation UTMAC allows file transfer to Macintosh and printing 2 31 7 7 Result files Select the automatic filing start and stop strings in the Filing dialogue A column selection marker data marker can be used with programmable output see 2 26 3 See the UTMAC User Manual for details Result files will be saved in the same directory where the current UTMAC setup document lies 2 31 7 8 Printer connection The MicroBeta program can automatically direct printer output through UTMAC However it is not printed immediately but gathered into a temporary printer output file You can print the output by pressing the Print button that appears at the top of the terminal window or by selecting Print Printer Output from the File menu After printing the contents of the temporary file will be deleted It is recommended that you print the printer output only when the counter is in the Ready state 2 31 7 9 Exit to Finder If you are running System 7 or MultiFinder you can always switch to other programs and let UTMAC work in the background If you want to stop counting temporarily type D see
112. assay batch When the end of the batch is reached counting starts from the beginning by counting the repeats and replicates again This procedure continues for as many times as you have defined for the cycle parameter Using injection When injection is selected it changes the counting procedure so that the positions are dispensed and counted multiple times depending on the number of Injector Modules used in the protocol Also the background sample measuring using the Background sample Y option is different the background is measured for every sample as the first repeat RP 0 using the counting time specified for the background sample The following example shows the order in which operation occurs Dispense with two modules background sample selected with two cycles two repeats and two plates Plate 1 For all coded positions Background Count with counting time of background repeat 0 Module 1 Dispense with Module 1 Wait the delay time of module 1 Count with counting time of module 1 repeat 1 Count with counting time of module 1 repeat 2 Module 2 Dispense with Module 2 Wait the delay time of module 2 Count with counting time of module 2 repeat 1 Count with counting time of module 2 repeat 2 Wait plate delay time Plate 2 For all coded positions Background Count with counting time of background repeat 0 Module 1 Dispense with module 1 Wait the delay time of module 1 Count with counting time of module 1 repeat 1 Count
113. assay protocols An 5 MultiCalc assay protocol tells which NV Workstation counting protocol MicroBeta is to use and what should be done with the data when it is obtained e g quality control analysis Terminal mode is available with MultiCalc and should be used if you want to edit normalizations standardizations and counting protocols from within MultiCalc With a terminal 2 37 you can get results to your screen print them out or store them on a disk in MicroBeta A terminal emulator 2 3 is a program which makes a PC act as a terminal You can have results stored on your PC You can also use your PC for other tasks during counting Injector assembly Results are sent Output Local Results go to fits here with the to MicroBeta to ext area PC connected module s below drive B during compu net to port 1 in Port 3 is used for terminal operat ter e g a work terminal connection to the ion For diskette main conn emulation amp injector modules information see frame ection MultiCalc See Settings Baud 6 2 10 Connect LAN 2 10 for Parity 1 Data bits to port using diskette info 8 Stop bits 2 2 or PC PC MultiCalc Handshake 1 see printout goes to 2J 30 For System setting info e g printer or printer connect Entering commands into terminal type 2 30 ed to the PC MicroBeta is described in 2 1 For Help type H h or 2 15 MultiCalc has its own manual 11 12 Par
114. ata Print normalization data Operations in the DPM Standardization Protocol State The same operations as in the Counting protocol state plus R eplot Plot DPM standardization curve Operations in the MultiCalc Assay Protocol State Contains only operations S how P rint and L ist Operations in the Conveyor State Help Display Help text Q uit Back to the Ready state Off Stop conveyor C lear conveyor Move cassette to rack and move rack to the middle Rack u p Move the rack up one level Rack d own Move the rack down one level Test level contains also Remove ca s s Move cassette to rack N ext cass Move next cassette to conveyor and read ID P rev cass Move previous cassette to conveyor and read ID Cu r rent cass Move current cassette to conveyor and read ID Ne x t pos Move cassette to the next position G oto pos Move cassette to a specified position C Dose detector Close detector block Op e n detector Open detector block Next le v el Move rack down to next measurement level 22 Reset r a ck Dnit mask Change m ask T est IDs F ocus ID reader J ust read IDs Print ever y ID Print ra w IDs Print b ad IDs Si z e checkpoint Calib 0 pos Calib det bloc k Park det Shelf 1 16 Calib sensors 2 1 Beginning operation of MicroBeta TriLux Move rack to the highest level Initialize detector mask small holes Change detector mask Read and print I
115. ation plate format See the next page for the examples of statistical output 2 29 3 Examples of statistical output The following protocol only relevant parts shown demonstrates the different statistical output formats 30 Printer output 1 No 2 SNORE 3 LONG 4 Programmable See 32 Display OuLpuL 1 No 2 SHOE 3 Long 4 Programmable 352 34 External output 1 No ZV SnOrT 3 Long 4 Programmable 4 gt 35 External output POS CTIME CCPM1 CCPM1 CCPM2 CCPM2 MEAN SEM1 MEAN2 gt SEM2 gt 36 File output 1 No Z2 Short 3 bong 4 Programmable 4 gt 37 Pile Oucpucs POS CTIME CEPMI CCPMIS se ECBM2 ECEPBMZS STAT NE MEAN lt 12 MEAN1 TSEM1 S SEM1 MEAN2 TSEM2S SEM2 gt File path ESXMBATES TI x gt File name extension gt 40 Change special features Y N gt 41 Counting control Y N Y gt Number of repeats gt Number of replicates I gt Number of cycles L gt 144 2 29 Statistics Because symbol S1 does not exist in any of the programmable lines 35 and 37 above output selections the deviation values are represented at the 2 sigma level The output defined by the protocol above for each output medium looks like the following EOS CTIME DOP UE CPM CCPM CCPM FLAG A01 939 Sisto me ZO To Zo Toad AO 112834 112834 ys A02 ee 3 0 S52 29997 ASE 208 110761 110761 NERS A03 oes oOo 32302 3423207 Zot LT L652 111652 US 3 replicates AQ1 A03 MEAN THEOR ERROR OBS ERROR 5 CHI SQUARED PROBABILI
116. ay Help text Back to the Ready state Initialize the selected injector module Volume to be dispensed via the selected tubing of the selected injector module Fill the selected tubing of the selected injector module Ensure there is no drop hanging from the selected tubing of the selected module Wash the selected tubing of the selected module with the selected volume of liquid Suck liquid in selected tubing of the selected injector module back into bottle 195 2J 1 Beginning operation of MicroBeta JET Co m mand S tatus Of Njext pos G oto pos C lear conveyor Disp e nser IN OUT Injector specific macros can be sent to the selected injector module Show the status of the selected injector module Stop injection Move the plate to the next position Move the plate to the selected position Return the plate to the rack Move the mask adapter with injectors injector assembly in or out Operations in the System State Help Q utt MS D O S E rror beep S tatus display V ersion C lock Se t up mode Setup mode contains also R S 232C P rinter D ata drive Ter m inal P a sswords Customi z ing Detector temp Test level contains also Serial n o Temp cali b Detector shelf u sage Display Help text Back to the Ready state Exit to DOS Set error sound on or off Set status display on or off Show version number of program and MS DOS Set date and ti
117. background sample question if you want to give a background value The value given in CPM is subtracted from the sample CPM for an isotope Two values are given 121 2 25 Protocols one for each window if dual label counting is used In dual label DPM counting three values are given The value is in the range from 0 0 default to 99 9 CPM or 999 999 9 LCPS Note in the normalization protocol press Y if you want to count the background plate 43 Half life correction Press Y 1f you want to use half life correction The half life for the isotope is displayed e g 107 500 hours for H No half life value is given for 4C because it is so long Note that you must always specify the half life if you have selected an isotope of type Other It must be in a range from 0 no half life to 999 999 99 hours 114 years If normalization or standardization protocol is selected on line 3 then half life is as specified in this protocol and it cannot be changed in the counting protocol Press Y on the next line if you want to set zero time otherwise the activity is corrected to the time when counting of the first sample of the assay was started The zero time consists of date and time First enter the date in the format DD MMM YYYY where DD is the day number 1 31 mum the month in letters JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV and DEC and YYYY the year e g 15 JUN 2000 The date can also be in the format YYYY MM DD e g
118. cale 32 Display output This output is selected when you want to display results on the screen It can be chosen independently of the printer output Display output has the same output alternatives as the printer output The default programmable output PRINTER means the same data as specified for printer output 119 2 25 Protocols 34 External output This output is selected when sending data to an external device such as a mainframe computer connected to the MicroBeta via port 2 The data is in ASCII format text files The output alternatives are the same as in printer and display output except that alternative 1 No output is the default value The default programmable output PRINTER means that the same data is specified as for printer output 36 File output The counting results can be stored on a floppy disk or hard disk on the PC or MicroBeta or on the server of a local area network LAN section 2 8 Datafiles The default setting is 1 no file output The alternatives are the same as in printer output The default programmable output PRINTER means that the same data is specified as for printer output The result files are named so that the first part of the name the part before the period is the protocol number and the second part after the period is a running number indicating the assay s execution order E g the first file for protocol 5 1s named 5 001 the second 5 002 etc Result file names for norma
119. can have None Odd or Even where None 1s the default 148 2 30 System The number of data bits can be 8 or 7 where 8 is the default The number of stop bits can be 2 or 1 where 2 is the default Lastly you can select the kind of checking process for the data transfer the handshaking It can be None DTR or XON XOFF where the default is DTR Confirm the selection of these parameters by typing Y twice When you have done this the program prints the original and the new settings and returns to the System state if serial port 2 or 3 setting 1s in question However when changing the terminal port port 1 setting the program then shows a help text that informs you to set your terminal to the same settings which were selected for MicroBeta before confirming the new setting When ready press any key The program should then ask Do you see this text Y N If the text appears it is quite sure that the communication works and you can press Y to confirm the change of terminal port setting If the text does not appear it is obvious that your new terminal and MicroBeta serial port settings differ In that case check from the printer the new MicroBeta setting and modify your terminal setting if required until you see the above text If you cannot make the communication work you have still one possibility Set the terminal to the original serial port setting Switch off the counter and switch on again Restarting MicroBeta at this phase returns the origin
120. cima1 gt The program adds an extra space character between printout fields lt decimals gt Number of digits to the right from the decimal point works for real number values such as CCPM1 only In the examples following character denotes a leading space character and it is supposed that the value of the CCPM1 field is 1234567 123456 Examples are given in the table at the head of the next column FORMAT QUTPUT EXPLANATION SPECTELEATION OBTAINED CCPM1 1234561 Default format CCPM1 7 0 CCPME T73 tel 23845674123 13 characters 3 decimals CCPMLE i334 UA SA OO AS ES Last digit 5 because of rounding CCEML TIA 123456071239 Exactly 12 digits in field CCPM1 11 4 WZ34567 123 Value too large strip off a decimal CCPM1 9 4 12345671 Value too large strip off 3 decimals CCPML 324 1234567 Value too large strip off all decimals CCPM1 7 4 1234567 Value fits exactly without decimals CCPM1 6 4 1 2E006 Value too large show value in scientific notation 1 2E006 means 1 2 times ten to the power 6 2 26 3 Entering constant text into an output Text can be defined between apostrophes The syntax 1s lt text gt or lt text gt lt width gt where lt text gt Arbitrary commentary text lt width gt Width of the text field If this is greater than the number of characters in lt text gt then leading spaces will be produced Examples SPECIFICATION OUTPUT EXPLANATION Example Example Character inside s shown Example
121. ckground samples Half life correction corrects the CPM DPM values for decay of short lived isotopes Zero time start of assay or specified date and time Chemiluminescence correction corrects the CPM DPM values for random coincidence contributions Statistics Monitor detects distorting contributions from static electricity Static electricity discharge 1s a phenomenon which occurs together with the use of plastic sample carriers such as microtitration plates 3 4 14 2 Normalization protocols Up to 100 Normalization protocols data sets Normalization of the detectors is required to eliminate possible difference in counting efficiency between the detectors Normalization is done for the different isotopes different applications or different window settings The Normalization protocol to be used is selected in the counting protocol Also available in the protocol are Half life correction Chemiluminescence correction Password protection 271 3 4 Specifications 3 4 14 3 Standardization protocols Up to 100 Quench standardization protocols data sets Quench standardization is needed to correct for counting efficiency variation or quench in the samples The Quench Standardization protocol to be used is selected in the counting protocol Selection of curve fit method smoothed spline interpolation spline linear interpolation or linear regression Input of individually adjusted DPM values for the standard samples Also ava
122. counting If N Next pos or E Next protocol is pressed the program starts to count the next sample position or the next assay respectively If quick view is used then N stands for the next plate Counting of an Assay protocol in MultiCalc is stopped by pressing the Fl key F2 and F3 are used for counting the next position or the next assay respectively 35 2 5 Counting 36 2 6 Counting control 2 6 Counting control See chapter 2J 6 for JET information 2 6 1 Counting parameters The counting process is controlled by the following parameters in the counting protocol 10 Counting time 12 Precision 2 sigma 41 Counting control Number of repeats Number of replicates Number of cycles Maximum channel counts a hidden parameter 48 Delay between plates Note Assay protocol parameters are described in chapter 2 20 MultiCalc A CPM normalization protocol or DPM standardization protocol does not include parameter 41 Counting control Instead the crosstalk normalization and crosstalk standardization protocols contain counting time for both crosstalk samples and standards e g 10 Counting time for crosstalk 11 Counting time for standards 2 6 2 Counting time Counting will be stopped after the specified counting time in seconds has elapsed from the start of the counting unless some other conditions have already stopped it The maximum counting time is 999 999 9 s more than 11 days 2 6 3 Precision 2 sigma The statis
123. cter in the template E g Control B is entered by first pressing down the Control key usually labelled with Ctrl and then typing letter B lower case or upper case does not matter If you keep pressing both the Ctrl key and letter B down for a while then the repeat function of the terminal sends several Control Bs to MicroBeta Note that Control B and Control N move only inside the default or the current value displayed before gt not inside a newly typed text The cursor control keys left and right arrows can sometimes be used instead of Control B and Control N This does not work with every terminal emulator The up and down arrow keys can be used when moving up and down in the protocol editor or you can type Lx to jump to line number x Please note that the Enter key must be pressed if making changes to any row As an example of keyboard editing suppose you have entered the following printout selection the second line Printer sOuLpat EOS CTIME ECEMI CCPM gt POS Cl IMR COUNTS CUPML CCPM IS After you press Enter you will see the text Error in output heading CTIMR and an arrow gt will appear Press the Control N keys As you do this you will see the letters POS CTIMR COUNTS I etc appearing After CCPM1 has been displayed the program will beep to inform that the end of the current value has been reached Keep Control B pressed down and you will see that characters at the end of output selection start disap
124. d gt given here Select crosstalk correction on line 3 Remember also to edit the parameters isotope counting time number of standards and the isotope activity See the listing above 2 7 3 4 DPM Standardization Counting Start the DPM standardization run in the Ready state or Count state using automatic counting and ID labels or by selecting Dx where x is the protocol number no ID label is needed in this case 46 2 Crosstalk correction The active samples are automatically counted first e g positions G9 B1 B11 and G12 then the crosstalk samples Al A11 H9 and H12 2 7 3 5 Counting the actual samples with crosstalk correction In the Ready state select a counting protocol to edit On parameter line 2 select DPM mode and on line 3 the crosstalk DPM standardization protocol defined above Start automatic counting using ID labels or start with the protocol number and press Enter The DPM values in the printout are the crosstalk corrected values 47 2 Crosstalk correction 48 2 8 Datafiles 2 8 Datafiles 2 8 1 Use Counting results can be stored on a floppy or hard disk located in the PC terminal emulation or MultiCalc they can also be sent to the server of a local area network LAN Results can also be stored on the MicroBeta disks in drive A or B Ifa VT52 or VT 100 terminal is used then results are stored on the protocol disk in MicroBeta drive B see 2 31 6 4 Terminal The data in the fil
125. d also in the short and long outputs and the DPM monitor in DPM counting At the end of this chapter is a list of monitors and flags with their output headings and possible values 2 21 2 Statistics monitor The purpose of this monitor is to warn of unacceptable count rate variation in a sample The sample count rate in different periods of the counting time is measured to determine the sample count rate variation The variation must be within the range expected from the statistical nature of radioactive decay If the variation is outside the range due to e g chemiluminescence the sample is recounted A maximum of two recounts are made before the results from the last recount are output The statistics monitor is not active unless it has been selected in at least one programmable output 2 21 3 DPM out of range monitor This monitor evaluates quench levels of samples in DPM counting and compares these with corresponding quench levels in the standardization This is done by measuring SQP I for a sample If it is not within the standardization range the monitor gives a DPM out of range message The DPM out of range monitor is included in the short and long outputs in DPM counting It is no longer in use if 1t has been removed from the outputs 2 21 4 LCPS out of range monitor This monitor evaluates the light intensity of samples in luminescence counting to inform of possible counter saturation The monitor is set to HIGH if counter saturati
126. d results are quantitative The P counting efficiency is around 70 and background less than 10 CPM Crosstalk values less than 0 05 are achieved The cassette consists of a cassette body and a lid In the cassette body there is a steel plate which is the same size as the filtermat The filtermat is aligned on the steel plate according to the edges and the lid is clicked on the filtermat so that the small hole is in the upper left corner the Al corner To count the P cassette in MicroBeta choose other isotope upper PMT usage and window 350 1024 in your normalization or counting protocol No crosstalk correction is needed If you use Windows workstation you can use the parameter P cassette 8 by 12 for P membranes or you can set up a new parameter with the settings previously mentioned 109 2 23 Normalization 110 2 25 Protocols 2 29 Protocols See chapter 2J 25 for JET information 2 25 1 Four types of protocols A protocol is a set of parameters which control the counting of samples There are four types CPM normalization protocol DPM standardization protocol MicroBeta Counting protocol MultiCalc Assay protocol A CPM normalization protocol contains parameters used when doing normalization see chapter 2 23 Normalization and a DPM standardization protocol contains parameters for standardization see chapter 2 12 DPM Standardization A MicroBeta counting protocol 1s used when counting unknown samples An as
127. den to make the editor shorter Press Ctrl K Ctrl and K keys pressed at the same time to hide the current line To reveal a hidden line go to the next line and press Ctrl U To unhide all hidden lines go to line selection line in the beginning of the editor and press Ctrl A Press and Enter when you are finished with your editing and want to save the changes made If you want to exit without saving the new values press and Enter 216 2J 25 4 Example protocol listing 2J 25 Protocols The example alongside shows a luminescence protocol listing The parameters are described in section 2J 25 7 Counting Protocol no zo due LesJuly 2000 10 54 59 Name Flash lumi CPM normalization protocol no 2 Wallac 1450 MicroBeta V4 4 S N 4500759 i PEOCOCOL name Flash lumi 2 Counting mode 1 CPM 2 DPM 1 2 CPM Dormalizearion protocol 4 Number of labels 1 Single 2 Dual 1 5 Isotope 1 LIAE AAA 3 C 14 43535 A pl 6 P 32 7 P 32 Cerenkov 8 Other 9 PMT use 1 Normal 2 Upper 3 Lower 2 Window 1 B L024 Use mask adapter Y N Y Use injector Y N Y Dispensing 1 Automatic 2 Manual L Dispensing 1 High 2 Low 3 Custom 1 Dispensing modules LL 7 Sample volume uL 100 8 Dispensing volume 1 uL alae Dispensing volume 2 uL 10 9 Delay time after dispensing 1 s 1 0 Delay time after dispensing 2 s 1 0 10 Counting time 1 s iene Counting time 2 s 1 12 Precision 2 sigma Daz 20 Count all positions in cassette Y N x 23 P
128. determining the efficiency for the particular detector The DPM value is counted by dividing CPM with the efficiency See chapter 3 3 Calculation methods for formulae used in DPM counting 2 11 3 Easy DPM Easy DPM is a special case of single label DPM counting Only normalization standards are counted in standardization and stored quench curves are used when counting unknown samples 61 2 11 DPM counting 2 11 4 Dual label DPM Dual label counting is similar to single label counting except that two plates are used one for each isotope The DPM1 and DPM2 values are calculated using the 3 2 method Note EasyDPM is not available with ParaLux count mode 62 2 12 DPM standardization 2 12 DPM Standardization 2 12 1 What is standardization When counting DPM samples in a multidetector counter the variation in detector efficiency detector energy and the effect of quenching must both be corrected for Standardization is the name given to the process in MicroBeta which makes this correction 2 12 1 1 Quench correction Several samples labelled with the same activity will give different count results even in the same detector because of the difference in the amount of energy absorbed in the samples This is called quenching To correct for this a number of samples with identical DPM but differing quench have to be counted in one detector This establishes a relationship between a quantity called SQP I or AQP I the latter is used fo
129. doing this then answer Y Yes Then MicroBeta stores a result file at first on the protocol diskette and sends it to the terminal as one block as Start Of assay ixXx Result data End of assay 239 2J 30 System where xx is the protocol number If your terminal emulator cannot capture data then answer N No Then the result files are stored on the protocol diskette of MicroBeta but not transferred to the terminal PC Note 1 Result file will be output if protocol line 36 FILE OUTPUT is set to other than 1 No output Note 2 See chapter Results for details 2J 30 10 4 Automatic result file deletion For VT52 and VT100 default Y Yes The capacity of the protocol diskette is limited 300KB 1 3MB depending on the number of protocols and it is capable of storing the results from at least 20 cassettes 1f the output selection for FILE OUTPUT does not specify too many different printout fields If this selection is set to Y Yes then the result files on protocol diskette will be deleted automatically when starting counting and after sending a result file to the terminal PC is complete If you want to ensure that your data has been transferred to another diskette or to some PC then answer N No In that case after starting counting the program confirms deletion before operation by asking Delete result files saved on protocol disk and you can answer N NO to reject deletion or Y Yes to accept deletion 2J 30 10 5 Print through termin
130. during counting and 1t should not normally be opened unless counting has been interrupted by pressing O If you need to add more cassettes then you can open the door and place the cassettes in a suitable empty place Close the door Counting of those samples which were in the counting position when the door was opened will be done again While counting is taking place the uncorrected CPM or CPS values are shown on the display of the Terminal PC These are replaced by CCPM1 DPM1 or LCPS values when the whole plate 34 2 5 Counting has been counted In addition there is the information from the barcodes on the cassette assay number protocol number cassette number function codes and the number of the shelf from which the cassette was taken There is also the current date and time for the assay the counting time set in the protocol and the current counting time in seconds for the samples in the measuring position If there is a specified number of samples given on line 22 in the protocol or normalization or standardization samples the assay end time and date will be given also During counting of an Assay protocol in MultiCalc live information from the counter 1s not normally shown However by pressing the F4 key LIVE some information will be displayed e g protocol No current and set counting times and positions that are counted 2 5 3 Stopping counting Counting can be stopped anytime by just pressing O off The program terminates
131. e 2 2 Robotic loading interface See chapter 2J 27 for JET information The robotic loading interface is a special door adapter system that brings a cassette outside so that a robotic arm can change the plate In the MicroBeta program the robotic loading interface is selected as follows S ystem L evel T esting Detector shelf u sage Use shelves 1 1 2 16 3 32 gt 1 L evel N ormal counting The operation is controlled using the RS 232C port 2 external port of MicroBeta When the cassette is outside MicroBeta sends the text Ready and carriage return line feed The possible commands are Hor list commands Nxx start normalization using protocol number xx Dxx Start standardization using prot number xx XX start counting using protocol number xx A start counting using IDs All Ready commands must be followed by a carriage return line feed or Counting can be interrupted with O After a plate has been measured and the cassette is outside MicroBeta sends the text Continue and carriage return line feed At this point O stops counting and any other character continues counting If plates are counted in the inverted position Al at the bottom right corner then Plate orientation Rotated should be selected in the counting protocol This selection reorders results so that outputs POS and SEQ are correct 139 2 2 Robotic loading interface 140 2 28 Safety and radioactive materials 2 28 Saf
132. e the normalization data will be destroyed when editing an old normalization protocol The normalization must therefore be done again for this protocol Only the protocol name and password can be edited without destroying normalization data See chapter 2J 25 Protocols for further details on normalization parameters and editing 2J 23 4 Preparing the normalization samples When injection is not used Depending on which type of MicroBeta you have it is supplied from the factory with for 1 6 detector instruments with 24 or 96 well capability a 24 well normalization sample plate containing standards for H and C The H standard is in D5 and the C standard is in D6 The sample positions and the activities for the standards are shown on a label on the plate The activity values can be inserted into the normalization protocol 209 2J 23 Normalization The standard 3 is Sample position in a normalization plate Be careful that the samples are in the correct positions when making your own normalization plates The positions cannot be changed by modifying protocol parameters In 24 well plates the standard for isotope 1 is in D5 and the standard for isotope 2 is in D6 Isotope 1 is the lower energy isotope in dual label counting In single label counting the standard is in the isotope 1 position 1 e D5 with one exception the standard for C must be in the isotope 2 position in single label counting with 24 well plates
133. e 400 uL well For aqueous samples SPA or for samples with Polystyrene Safe cocktail 1450 444 Sealed with tape type 1450 461 or 1450 462 Counted in cassette type 1450 103 1450 408 24 well Heat sealable Sample Plate 25 box 24 flat bottomed wells 4x6 format made of clear PET plastic printed lines between wells chemically resistant to all HiSafe cocktails max volume 1 ml well Sealed with tape type 1450 461 or 1450 462 or heatsealed with 1450 463 in 1495 021 Microsealer using 1495 023 Heat Seal Support 24 Counted in a cassette 1450 102 1450 410 96 well Rigid Sample Plate 25 box 96 flat bottomed wells 8x12 format made of polystyrene white well walls and clear well bottom max volume 350 uL well For aqueous samples e g SPA or for samples with Polystyrene Safe cocktail 1450 444 Sealed with tape type 1450 461 or 1450 462 Counted in cassette type 1450 105 1450 419 ScintiStrip 120 box 12 breakable strips of 8 wells on a support frame 1450 482 made of scintillation plastics max volume 300 uL well For immobilised samples or aqueous samples no scintillation cocktail needed Sealed with tape type 1450 461 or 1450 462 Counted on the support frame in cassette type 1450 105 1450 514 96 well Isoplate 25 box 96 flat bottomed wells 8x12 format made of clear polystyrene wells with a rigid white frame max volume 350 uL well For aqueous samples e g SPA or for samples with SuperMix cocktail 1200 439 Sealed w
134. e are anticipated a mains stabilizer may be necessary 287 4 1 Installation instructions 4 1 3 Unpacking 7 Removing ANS MicroBeta from A 1ts packaging a g lt Cut off and remove the binding bands Open the clinching nails and lift the cover off from the base of the package see the figure in the previous column Unpack all units and accessories and check them according to the packing list also noting any possible transport damage Open the lower clinching nails and remove the packing around the main instrument Lift the instrument up from the bottom of the package Move the instrument to its place of operation 288 4 1 Installation instructions 4 1 4 Checking the mains voltage setting Measure and note the mains voltage at the outlets to be used Locate the mains selector switch this is on the right side when looking from the rear of the instrument If necessary adjust the mains selector switch to correspond with the measured supply For supplies with a nominal voltage of 230 V it is recommended that the selector be set to 240 V Check that the fuses fitted in the fuse carriers on the back panel are of the correct rating for the local supply and according to the label T 4 A for supplies with a voltage in the range 100 to 120 V and T 2 A for supplies with a voltage in the range 220 to 240 V For continued protection against fire hazard replace only with the same type and rating of fuse Insert the batter
135. e below for details and chapter 2 30 System Terminal 2 31 6 2 Special features The terminal understands standard VT52 VT100 escape and other control codes 2 31 6 3 Restrictions If the product does not understand VT52 VT100 block graphics characters then set System Ter m inal Block graphics to N No 2 31 6 4 Result files There are several possibilities 1 MicroBeta automatically saves results defined on the Protocol 36 File output line into directory B RESULTS on the protocol diskette You can copy files from there if your PC can read standard MS DOS 3 5 inch 1 44MB diskettes 2 If you set System Ter m inal Send result files to terminal to Y Yes then MicroBeta sends the result file saved as in 1 at the end of each assay as one block of text in the format otart OL essay xx End of assay where xx is the protocol number The terminal emulator can now capture the data within Start of assay and End of assay 3 It is possible to define the following environment strings in the MicroBeta AUTOEXEC BAT SET SOPN lt open gt open result file name follows SET SCLS lt close gt close result file SET SBEG lt begin gt save characters until lt end gt is found SET SEND lt end gt end character saving 169 2 31 Terminal emulators In the above lt open gt lt close gt lt begin gt and lt end gt can be any character sequences which do not include the VT52 VT100 control codes and do not appear i
136. e format can be selected by including the keyword PLATE in the programmable output selection of the counting protocol The user must then also select the printout fields of interest Each printout field such as ccpm1 will be output as its own table of 6 or 12 columns and 1 8 rows MicroBeta outputs all the selected tables after the cassette has been counted The output can contain both normal and plate format output Items before the PLATE word are output in normal format and those after it in plate format Note If you are using MultiCalc Assay protocols you can also make plate maps there See section 2 22 4 2 and the MultiCalc manual chapter on Plate operation Plate mapping is done in a MultiCalc Assay protocol as part of the CODING section F6 PLATE is the softkey to press 2 19 3 Restrictions Some restrictions exist caused mainly by memory requirements Plate format cannot be used if the Repeats parameter is greater than 1 Plate format cannot be used if the spectrum plot symbol SPECTRA has been selected for printer output Plate format should be selected for the Display only if results are filed with UltroTerm In this case the plate format should come between the data markers PS and PE A maximum of 4 different printout fields will be accepted resulting in up to 4 tables If plate format is selected for more than one output medium display printer external result file then the printout fields listed with the first
137. e maximum number of replicates are shown below If there is less than the maximum number of replicates the replicates positions are from left to right and top to bottom 3 and 6 det models without background samples Rows E H are empty in the 3 det model l 2 3 4 5 6 y 8 9 10 11 12 3 and 6 det models with background samples Rows E H are empty in the 3 det model T0 0 40U00 Y42 gt 1 2 3 4 5 6 7 8 9 10 11 12 B Si s1 si Si S2 s2 82 82 S3 S3 s3 S3__ D Si si si SI S2 s2 82 S2 S3 S3 3 S3 E B4 s4 s4 S4 B5 S5 S5 S5 B6 s6 s6 s6 Est S4 St 154 85 185 85 85 86 86 86 186 m fst s4 s4 tst ss 55 85 ss 56 se s6 se l and 2 det models without background samples Rows E F are empty in the 1 det model e a a A Sl O SS TO A B fs s Ys Si e e C All ft S S T S F s2 s2 s2 8 S S T Go A O ff ff a a 0 So se ss 203 2J 18 Luminescence counting l and 2 det models with background samples Rows E F are empty in the 1 det model TR O FE E E O E CA E EC FE E E a a a ff POSO AOS IO IO OS IS IA IS IS IE FE E ECO O O O O O O O O O IS IS 2J 18 5 Preparation of flash type normalization samples 24 well format Plate maps for different detector configurations with 24 well plates and the maximum number of replicates are shown below If there is less than the maximum number of replicates the replicates positions are from
138. e of ID labels Delay Start allows for sample incubation prior to start of measurement Edit Copy Print Delete List features for management of protocols Password protection against unwanted changes of protocol Help function built in operating instructions supports the user throughout the program Single label CPM or DPM counting Dual label CPM or DPM counting Crosstalk correction to allow different types of microtitration plate to be used Scintillation proximity assay can be done Preset parameter for 6 common isotopes Count termination by counting time or reached precision 2 sigma error Plate counting control whole plate or only active rows or positions Default Short or Long printer output for quick printout settings Fully programmable printout selection Including statistical analysis of results and user functions Fully programmable output to display Including statistical analysis of results and user functions 270 3 4 Specifications Fully programmable output to mainframe Including statistical analysis of results and user functions Fully programmable output to file on terminal PC Including statistical analysis of results and user functions Repeat or Replicates counting Cycle counting repeat counting of the whole assay Background correction CPM DPM results can be corrected for experimental background either by typed in background value or background samples Programmable counting time for ba
139. e operated with MultiCalc Now with this instruction you are telling which counter you are actually going to work with If MultiCalc cannot get MicroBeta into the Ready state it will ask you to press F3 to get into the terminal mode Then press F3 and try to get MicroBeta into the Ready state If communication does not seem to work then see chapter 2 31 4 11 Terminal emulators GenTerm Trouble shooting 1 about how to try to fix the problem You can also try to press the Esc key several times At the end of installation answer YES to see instructions about MultiCalc MicroBeta use Note Steps b through f should be executed when MultiCalc 1s connected for the first time to MicroBeta MicroBeta has been used in another environment such as with UltroTerm There seems to be some problems For more details about MultiCalc installation see the MultiCalc User Manual Installation 2 22 3 MultiCalc terminal mode Sometimes you must use terminal mode e g when changing counting standardization or normalization parameters MultiCalc allows you to operate in terminal mode as mentioned in the previous section The type of terminal emulation used by MultiCalc is W152 Le when MultiCalc is in terminal mode MicroBeta sees it as if it was a VT52 type terminal MultiCalc must therefore be selected as terminal emulation in the MicroBeta System mode parameter Ter m inal see chapter 2 30 10 System Ter m inal 96 2 22 MultiCalc operatio
140. e other emulation see 2 31 4 11 Troubleshooting below If there still seems to be communication problems after changing to the same emulation in both the counter and the PC the reason can be that there are different communication parameters in the counter and the emulator Here is a quick guide for how to change the communication parameters in GenTerm For further information see the GenTerm User Manual 1 Press the Ctrl and Break keys to get to the GenTerm main menu or in VT 52 mode press F8 2 Use the down arrow key to go to Set terminal parameters and press Enter 3 Select an item from the Set Up menu by pressing the up or down arrow keys 4 Change the value of the selected item by pressing the left or right arrow keys 5 Store the setup on disk as default settings by pressing F3 6 Press F9 to return to the terminal emulator mode Section 2 30 System R S 232C in this manual describes how to change communication parameters in the counter This section also contains the default parameter settings 2 31 4 6 Special features GenTerm WT emulation emulates the IBM ROM BIOS keyboard and display functions If communication problems arise see the Troubleshooting section below 162 2 31 Terminal emulators 2 31 4 7 Restrictions Do not change GenTerm to ADM 3A or VT52 emulation 2 31 4 8 Result files The destination disk drive and directory can be set in System D ata drive or in Protocol File output The MicroBeta program u
141. e protocol number area and with a NORM label in the function code area Insert the cassette into the rack and close the door Press either A Automatic counting or Nxx where xx 1s the normalization protocol number in the Ready or Count state to start counting If Nxx is selected the program asks Do you want to continue counting after normalization Y N gt If Y Yes 1s selected the counting continues after counting the normalization plate if N No is selected the counting stops after the normalization If normalization has already been done for the protocol then the program asks first This protocol contains normalization data Continue Y N gt This is to prevent accidental overwriting of normalization data If automatic counting is selected the normalization cassette can be placed anywhere in the rack The NORM label is needed in this case The counting results for the normalization are output as specified on lines 30 37 in the normalization protocol and after them the background and detector efficiency values are given Note Normalization and unknown samples should have the same quench level in dual label counting 2J 23 6 Preset normalizations There are four preset normalizations available in MicroBeta Normalization protocol 1 contains a H normalization using ScintiStrip plate 1450 419 dry 200 uL volume with crosstalk correction Normalization protocol 2 is the same as but using ry Normalization prot
142. egister is provided for each of the 7 standard isotopes and one register for the user definable isotope called Other At the end of each assay a total accumulated count rate or activity for that assay is printed out Each of these count rates is then accumulated to form the Total count rate This is then the total count rate for all assays since the last time the total count rates were reset You can display the total count rates print or reset them You can also define efficiencies for each isotope these are used to transform count rates into activities Type T in the Ready or Count state to get to the Total count rate functions See the example on the next page 177 2 33 Total count rate Count H elp ONU TE S how Print R eset D efine efficiencies Total Count rate S Total Count rate and activity since Wed 1Z2 July Z000 132535 Isotope CCPM DPM BS LID TIO 0 L TZ5 0 0 C 124189482 297103992 S 35 86669 0 Creo 0 0 P 32 0 0 P 320 0 0 Other 2014554 0 H elp Q uit S how P rint R eset D efine efficiencies Total countrate gt D Efficiency estimates used Y N N gt Y Isotope Efficiency LT H 3 507 00 gt 2e ESEAS 0 00 gt 3 E lA4 75 00 gt 90 A S30 0 00 gt iS 0 00 gt 6 P 32 0 00 gt Y P 32C 0 00 gt 8 Other 0 00 gt H elp Q uit S how P rint R eset D efine efficiencies Total Countratle gt 5 Total countrate and activity Since Wed 12 July 2000 13253 Isotope CCPM Eff DPM ne
143. en if the display is selected If the number of cassettes is defined the estimated end time is also displayed 28 2 4 Coding cassettes and filtermats 2 4 Coding cassettes and filtermats 2 4 1 Cassette code system Barcode labels are used for cassette and sample recognition They are attached to the marked area of the ID support plate This is a separate piece of plastic which can be clipped onto the appropriate area of a cassette as shown in the figure When fixing ID labels on the ID support plate take care that the fixing area is clean Sometimes it happens that the protocol ID label is dirty or placed the wrong way round and the barcode reader fails to read it In this case the default protocol prot No O is used 2 4 2 Cassette codes used An ID support plate has four fields areas to which a barcode can be fixed The meaning of each field is described below The figure shows a counting cassette The cassette number is 1 and the counting protocol to be used is 60 ounting norm or stand protocol Mo Multivalc Assay protocol Mo ID support plate and barcodes 2 4 2 1 Function codes The FUNC field can be labelled as follows No label The cassette is a normal sample cassette without any special function STD The cassette is to be used for DPM standardization See chapter 2 12 DPM Standardization The number of the standardization protocol is given in the PROT field 29 2 4 Coding cassettes and filtermats
144. er has been switched off normally only when the instrument is installed Details of setting the clock are given in chapter 2 3 Clock When you set the clock the Terminal PC date and time are also set if GenTerm is used Example 5 system gt C Date 12 Jul 2000 gt 13 Jul 2000 Time 14 44 gt 9 30 Set clock Y N gt Y 2J 30 7 R S 232C There are three RS 232C communication ports which can be used 1 for the Terminal PC 2 for an external computer 3 for injector module communication You can specify the communication parameters for each of these ports separately Give the number of the port and press Enter The first parameter that can be changed is the speed of transferring the data the Baud rate The possibilities are 300 600 1200 2400 4800 or 9600 234 2J 30 System The default value for the terminal and external ports is 9600 For the printout port it is 4800 If these values are not suitable then select the ones you need Next is the parity selection you can have None Odd or Even where None 1s the default The number of data bits can be 8 or 7 where 8 is the default The number of stop bits can be 2 or 1 where 2 is the default Lastly you can select the kind of checking process for the data transfer the handshaking It can be None DTR or XON XOFF where the default is DTR Confirm the selection of these parameters by typing Y twice When you have done this the program prints the original and the new settings
145. ers is to be used for counting the samples in a cassette These identification labels are printed with barcodes and the reader is situated in front of the detector block There are identification labels for identifying the protocol and the cassette number There are also three special function codes STOP NORM and STD When a cassette carries a STOP code it is counted normally and after it the counting stops NORM and STD show that the cassette carries normalization or standardization samples respectively See chapter 2 4 Coding the cassette for more details You can also identify the plate itself with a barcode see section 2 4 6 Plate ID reader 3 1 6 Detectors One two three six or in the case of MicroBeta TriLux twelve pairs of photomultiplier tubes view the samples in the measuring position The detectors are placed in a 1x1 2x1 1x3 2x3 or 3x4 formation respectively see chapter 2 9 Detectors for pictures of this formation The tubes of one pair are situated on opposite sides of the counting block one above and one below the block Each pair operates in coincidence mode which greatly reduces single photon background events The out of coincidence signal is also measured simultaneously When a cassette moves into the counting position the labels on the cassette are read by the barcode reader Then 1 2 3 6 or 12 samples on the cassette are counted simultaneously according to the protocol specified by the label on the cassette Due to
146. ers in a protocol 114 218 Parity 148 235 Parking detectors 300 Password 122 155 156 227 241 246 Pause Data transfer 75 Printing 75 Performance 268 Plastic bags 274 Plate coding MultiCalc 100 Plate format Microtitration plate 83 Plate ID reader 32 268 PMT use 115 219 Polystyrene Safe 274 Port 1 49 Port 2 49 Ports 148 234 Positions 74 Power 265 Power failure 69 Power supply fan 253 PREC 94 Precision 37 116 197 221 Preset isotope protocols 104 209 Prime 184 Prime command string 245 Print through terminal 101 154 240 Printer 49 150 236 Connections 298 Output 117 145 222 Port 3 49 Type 298 Printing Pause in printing 75 Printout 5 Index Changing field format 132 Constant text 133 Default formats 136 Defining new fields 133 Items 127 Program failure 69 Protocol name 114 218 Protocol number 30 Protocols 111 215 Copying 114 218 Creating 112 216 Deleting 114 218 Editing 112 216 Listing 111 215 Parameters 114 218 Protecting 114 218 Types 111 215 Q Quench series preparation 88 Quenching 63 Quick view 116 Quick view 115 218 Quit 156 245 Radiation Window 179 Raise detector 243 Repeat counting order 38 With injection 199 Without injection 198 Repeats 38 198 Replicates 38 198 Results 127 File 51 Storing 49 Return key 16 188 Robotic loading 139 231 Plate orientation 117 222 Robotic loading interface 25
147. es saved on the disk is available to be processed or printed Data can also be sent to an external computer The data is sent via port 2 on the back of the counter Note Port 1 is for the Terminal PC The third output port 3 is for the printer 2 8 2 Disk drive selection The disk drive of the terminal PC where the data is to go can be chosen in the System state by pressing D Data drive You can choose from drives A B or C where C is a hard disk drive and A and B are for floppy disks You can also specify the whole directory path see the chapter System D ata drive Start the path with if you wish to store results on the MicroBeta disk Depending on the type of PC you have to proceed as follows Single floppy disk PC You must select drive A Then you must replace the terminal emulator program diskette with an empty but formatted data diskette Dual floppy disk PC Select drive B This allows you to keep your terminal emulator diskette in drive A Load an empty formatted data diskette into drive B Hard disk PC Select drive C hard disk or specify the path This will allow you to collect large amounts of data If you have only specified the drive you must select the directory on the terminal PC where you want to store your data To do this exit from the terminal emulator program See the chapter Terminal emulators Exit to MS DOS Use the change directory command in MS DOS 1 e cd directory name to go to the directory you wan
148. es symbol S1 in one of the programmable output selections then values will be output at the 1 sigma level 2 29 2 Restrictions If the user selects programmable output and includes any of the following statistical values except Sl and STAT MEAN CV1 SEM1 TCV1 TSEM1 CHISO1 P1 SD 1 SEM1 TSD1 TSEM MEAN2 012 SEM2 TCV22 TSEM2 CHISO2 P2 SD2 SEM2 TSD2 TSEM2 REPI 143 2 29 Statistics then a default statistical table summary will not be produced Instead you must specify in the programmable output selection which statistical values to output how to locate these values in the output and which kind of formats are to be used to output the values Statistical values cannot be used in arithmetic expressions The default values will be output at the 2 sigma level unless symbol S1 exists in the selection The external output of the example in 2 29 3 demonstrates a compact statistical output where only values MEAN1 SEM1 MEAN2 and SEM2 have been selected They have been output on the same line as the last replicate in order to reduce paper consumption The file output of the example in 2 29 3 demonstrates the use of the control symbol stat The output values defined after STAT will be output only when the last replicate or repeat result has been output This can be used e g to output the statistical data on its own line as in the example following or to produce replicate mean values when using Microtitr
149. estart MicroBeta by putting power off and then on again Terminal type baud rate and data stop bits are set automatically after instrument restart see 4 7 2 and 4 7 3 Another method is if the instrument is in the Ready state to press several times the key that sends the Escape character to MicroBeta usually labelled Esc in UltroTerm F3 MicroBeta should respond Please check terminal type in S ystem Ter m inal 163 2 31 Terminal emulators After doing this the keyboard will work and you can then set terminal emulation in System Ter m inal If the message above does not appear then the problem is probably in the serial communication cable RS 232C protocol 2 Printer errors GenTerm shows an error window Solve the printer problem power off off line mode paper out etc and then press R Retry to continue printing If you cannot fix the problem press A to abort printing and then P until the MicroBeta program asks Pause printing Y N Then answer Y 3 Diskette and hard disk problems When the diskette or hard disk becomes full GenTerm shows a message window that enables you to a Change diskette insert a new formatted diskette the result file continues on the new diskette with the same name b Give a new file name e g direct the result file to another disk drive c Stop logging data Select the recovery method using the left and right cursor keys press Enter and follow the instructions If the resul
150. et r a ck Dnit mask Change m ask Dest IDs F ocus ID reader J ust read IDs Print ever y ID Print ra w IDs Print b ad IDs Si z e checkpoint Calib 0 pos Calib det bloc k Park det Shelf 1 16 Calib sensors 2J 1 Beginning operation of MicroBeta JET Move rack down to next measurement level Move rack to the highest level Initialize detector mask small holes Change detector mask Read and print ID of each cassette continuously Focus and stop with ID reader on Read IDs continuously and show IDs on the screen Toggle printing of ID bitmaps in ordinary form Toggle printing of ID bitmaps in raw form Toggle printing of bad ID bitmaps in raw and ordinary form Move cassette to cassette type checkpoint Move cassette between zero point and D06 Move detector block up and down at mask change position Park detector block for shipping Move rack to the specified shelf Calibrate sensor offset steps Operations in the Total Count Rate State H elp Q uit S how P rint R eset D efine efficiencies Display Help text Back to the Ready state Show the accumulated total count rate and activity Print the accumulated total count rate and activity Reset the accumulated total count rate and activity Define counting efficiencies of the isotopes Operations in the Injector State Help Q utt Dnit D ispense P rime T ip Wash Suck b ack Displ
151. ety and radioactive materials The following comments about precautions and safety measures in handling radioactive materials are included as a guide and are not intended to be fully comprehensive More complete details may be found elsewhere for example in the booklet SAFE HANDLING OF RADIONUCLIDES published by the International Atomic Energy Agency Vienna this may be recommended as a useful code of practice appropriate to radio chemical laboratories Unless a specially designed radio isotope laboratory is used limitations should be placed on the amount of active material in the laboratory area depending on toxicity and type of chemical operation For high toxicity material and wet chemical operations involving the risk of spillage the AEA recommend a maximum activity of about 10 uCi Personnel should be properly trained in the safe handling of these materials maximum levels of stored activities should be set proper records should be kept and a definite monitoring schedule maintained The areas where samples are handled should be kept clean and free of dust This is most easily accomplished if all surfaces are as smooth as possible and 1f the minimum number of extra surfaces 1s introduced into the room Lastly it is extremely important to store all radioactive materials in a separate room to which access is restricted 141 2 28 Safety and radioactive materials 142 2 29 Statistics 2 29 Statistics 2 29 1 When statistical outpu
152. ever y ID Print ra w IDs Print b ad IDs Si z e checkpoint Calib 0 pos Calib det bloc k Park det Shelf 1 16 32 D i spIn Di s pOut Calib sensor Sensors gt H elp Q uit Y Rack X Rack XY Z ero XY Pos F ine D etUp Det U pPos P arkDet E dit 4 1 Installation instructions 4 1 8 Setting the system parameters During the installation some system parameters should be checked and set if required These parameters can be set in the System state of the instrument program see chapter 2J 30 System A checklist of the system parameters 1s provided in what follows 4 1 8 1 Terminal emulation Instrument Manual See chapter 2J 30 System Ter m inal Default UltroTerm terminal emulator program 23 lines on screen Depending on which terminal emulation the user wants to use check the following 4 1 8 1 1 GenTerm in WT emulation GenTerm is a terminal emulator program supported by MicroBeta The following parameters must be set if GenTerm is to be used a Set Terminal emulation to 1 GenTerm b Set Number of lines on screen at maximum to 24 Notes 1 GenTerm V2 B or later is required 2 See chapter 2 31 Terminal emulators GenTerm Installation 3 See also 4 8 2 and 4 8 5 4 1 8 1 2 UltroTerm This is the default terminal emulation and so normally you need not change the terminal emulation settings However if the terminal PC is some portable model that is not capable of displaying
153. fault formats of output fields The default format of printout fields is shown in the list here Note Fields are always separated by one space that is not included in the field length Note in these tables y stands for 1 or 2 or 3 and z for 1 or 2 a Sample identification data AS 2 XZ CA 4 XXXX NOS XXX CTED XXX Dz XX FU 4 XXXX PLATEID 12 XXXXXXXXXXXX POGES XXX PR 2 XxX REZ XX Spal X SEQ 4 XXXX SH 2 XX b Sample counting data AOP IJ 621 XIXX X AQP I 7 1 XXXXX X CCPMy 720 XXXXXXX CCPEMy T 61 XXXX X CEST ELA OO CLM 6 1 OEA COUNTSYS 7 XXXXXXX CPMy 7 0 XXXXXXX CPMy 6 1 XXXX X CPM AOP 7 0 XXXXXXX CPM C 7 0 XXXXXXX CPM NC 7 0 XXXXXXX Boyd XXXXXXX X CTIMESSsL PO e DATE 11 XX XXX XXXX DCEPMy 79 el XXXXXXX X DDPMz 9 1 SKK X DLCPS 9 1 XXXXXXK K DPMz 7 0 XXXXXXX DPMzS 3651 SOO aC DPMM 4 XXXX EFFy 6 4 O ETIMEZ V933 XXXIX XXX FLAG 4 XXXX PSE 6 1 SOO LE LOPS fk XXX X 136 2 26 Results LCPS 941 XXXXXXX X LCPS32 62 1 DOS LM 4 XXIX RESP zZ 9 31 EXXERXIX Xx RESPz 3 6 1 SOO o SOP CL S OSL XXXIX cK SOP I a TEL sexx x STIME 8 lt x PC STM 4 XXXX LIME 8 So Se UN Wy 232 XXXX X XXXX X c Statistical data CATSOZ OS XXIX 4 CV ze 622 XXXX X MEANZ 9 1 ROR RK XX POS SOCK x REPL XXX XXX ODZ ol XXXXXX X SEMZ 26 271 XXXXXX X SEMz 6 1 SOK LEVAS EOL XXXX X LoD Zo LI XXXXXX X TSEMzZ3 6 1 MK TSEMZ soe AI 137 2 26 Results 138 2 2 Robotic loading interfac
154. g 5 6 7 equals 000011100000 1 equals 100000000000 and 3 equals 001111111111 After pressing Enter you will be asked for positions for the next row The default values are the same as you have given on the previous row Press when you want to stop specifying counting positions The rest of the rows will then be counted as the last edited row In the example above the following positions will be counted for cassette 1 A3 A4 A5 A6 B5 and B6 The following cassettes can then be coded in the same way as the first one Press once again to exit cassette coding If there are more cassettes in a batch than there are defined cassettes the rest of them are counted according to the last cassette specification W can be pressed to code additional cassettes later X can be pressed to change the cassette type 96 to 24 or 96 to 384 and vice versa for cassettes 2 to 16 see the example 23 Plate orientation Select 2 if rotated plates Al at the bottom right corner are used 3 if mirrored Al at the top right corner and 4 1f both rotated and mirrored Al at the bottom left corner 117 2 25 Protocols 30 Printer output There are four printer output alternatives 1 No 2 Short 3 Long default 4 Programmable 1 No If No is selected no results are printed 2 Short If Short is selected sample results are printed using the short results format POS CCPM1 CCPM1 In dual label ccpm2 and ccpm2 are added ccpM is re
155. g by selecting protocol from list Nnn norm prot no Start CPM normalization by giving protocol number N Start CPM normalization by selecting protocol from list Dnn std prot no Start DPM standardization by giving protocol number D Start DPM standardization by selecting protocol from list A utomatic counting Start automatic counting using IDs Operate con v eyor Conveyor operations T otal count rate Show and reset total counts M anual counting Start manual counting only at Test level Operations in the Protocols State H elp Display Help text Q uit Back to the Ready state C ounting protocol Edit counting protocol CPM n ormalization prot Edit CPM normalization protocol DPM s tandardization prot Edit DPM standardization protocol M ultiCalc assay protocol Show and print MultiCalc assay protocol 21 2 1 Beginning operation of MicroBeta TriLux Operations in the Counting Protocol State Help Display Help text Q uit Back to the Ready state E dit Edit or create and edit a protocol nn prot no Edit a protocol by giving protocol number D elete Delete a protocol S how Show protocol parameters Print Print a protocol L ist Print list of protocols De fault Give default values to a protocol C opy Copy one protocol to another Operations in the CPM Normalization Protocol State The same operations as in the Counting protocol state plus N ormalization d
156. g features such as isotope or window are changed When a new type of microtitration plate is used crosstalk normalization may be necessary 103 2 23 Normalization 2 23 3 Editing normalization parameters Normalization parameters are stored in normalization protocols These parameters are shown in the figure There can be a maximum of 100 normalization protocols numbered from 0 to 99 Protocol number 0 is the default protocol fine levt4o type 7 to exit Y for Herp 1 gt 1 Protocol name gt 3 Crosstalk correction Y N N gt 4 Number of labels 1 Single 2 Dual 1 gt 5 Isotope 1 Ly ASS 2 tA SEELA Ay Sa 35 Cra 16 P37 7 P 32 Cerenkov 8 Other 9 Luminescence 1 gt ParaLux Y N N gt PMT use 1 Normal 2 Upper 3 Lower 1 gt Window 1 gt 360 10 Counting time for crosstalk s 60 0 gt 12 Precision 2 sigma 0 2 gt 30 Printer output 1 No Z Short 3 Long 4 Programmable 3 gt 32 Display output L NO 2 SHOE 3 Long 4 Programmable 3 e 34 External output 1 No Zi Shore 3 Long 4 Programmable L eae 36 File output 1 No ZV SHO 3 Long 4 Programmable di gt 40 Change special features Y N Y gt 41 Isotope activity setting Y N Y gt Isotope 1 activity DPM Z00000 r0 gt 150000 42 Background sample Y N N gt 43 Half life correction Y N N gt 44 Chemi luminescence correction Y N N gt 45 Use password Y N N gt 46 Special plate 1 Wallac 2 BP filter 5 OEneL 1 gt
157. g standards for H and C The H standard is in G10 and the C standard is in G12 for other configurations of MicroBeta special arrangements are made The standard 3 is Sa A Pm de hea si et a e a Sample position in a normalization plate The sample positions and the activities for the standards are shown on a label on the plate The activity values can be inserted into the normalization protocol 105 2 23 Normalization Be careful that the samples are in the correct positions when making your own normalization plates The positions cannot be changed by modifying protocol parameters In 24 well plates the standard for isotope 1 is in D5 and the standard for isotope 2 is in D6 Isotope 1 is the lower energy isotope in dual label counting In single label counting the standard is in the isotope 1 position 1 e D5 with one exception the standard for C must be in the isotope 2 position in single label counting with 24 well plates This is because the factory made normalization plate could be used for both single label H and 14C normalizations and dual label H C normalizations For best results 96 well plates should be used as normalization plates if 96 well sample plates are to be counted in a 1 6 detector counter The isotope 1 standard should be in G11 and the isotope 2 standard in H12 See 2 7 2 Crosstalk CPM normalization for crosstalk sample positions and 2 18 for luminescence sample positions
158. gt gt gt gt gt gt gt gt gt gt gt 2J 30 System This is the plate that fits between the detectors and the microtitration plate The number of holes in the plate correspond to the number of detectors This parameter allows you to select 1f the mask adapter is to be used It must be used 1f you want to use the injector system since the injector tubing is fitted into the mask adapter 2J 30 14 2 Injector module This parameter must be Y if you want to use injection 2J 30 14 3 Raise detector This parameter must be Y if you want to use injection because the upper detector must be raised to make space for the mask adaptor and injector s 243 2J 30 System 2J 30 14 4 Multichannel pump There can be from 1 to 6 channels for the pump If the pump is a single channel one 1 e for a single detector MicroBeta JET then select N here Otherwise select Y for a 2 3 or 6 channel pump 2J 30 14 5 Number of channels Give here the number of channels 1 2 3 or 6 2J 30 14 6 Number of injectors A single channel system can have up to four injectors whereas a multiple channel system can have a maximum of two injectors Give the number for this parameter 2J 30 14 7 Injector 1 As many injectors will appear as there are selected Each injector has its own set of paramters as follows 2J 30 14 8 Syringe volume This is the amount of liquid a syringe takes in during one stroke There are three possibil
159. hanged in the counter 1 Switch on the printer 2 Switch on the PC 3 Install UltroTerm as described in the UltroTerm User Manual and in section 2 31 5 3 Installation in this manual 4 Start UltroTerm as described in the UltroTerm User Manual and in section 2 31 5 4 Starting UltroTerm in this manual 5 If necessary change the communication parameters as described in the UltroTerm User Manual and in section 2 31 5 5 in this manual 6 Insert the MicroBeta program disk into drive A the lower disk drive of the counter 7 Insert the MicroBeta protocol disk into drive B the upper disk drive of the counter 8 Switch on the counter with the power switch on the back of it 9 After about 3 minutes Ready is displayed and the counter is ready for operation 290 4 1 Installation instructions 4 1 6 2 Start up using another terminal emulator When the program is started it will check what kind of terminal or emulator is connected to the counter and change its own settings according to this It will also test the baud rate and the number of data bits and make necessary changes The program can identify the same emulators as are available in the counter 1 e GenTerm WT UltroTerm MultiCalc VT52 standard VT52 and VT100 It can also identify baud rates from 300 to 9600 bps and 7 or 8 data bits Together with changing emulator and communication parameters the program will also notify the user that the change has been made and that the u
160. hase of entering a long input such as output selection in the protocol editor you can press Enter to complete entering a new value and then go back to the same line to correct the mistake 188 2J 1 Beginning operation of MicroBeta JET The editing keys are lt or Control B Go back one character in the template default or current value gt or Control N Go to the next character in the template E g Control B is entered by first pressing down the Control key usually labelled with Ctrl and then typing letter B lower case or upper case does not matter If you keep pressing both the Ctrl key and letter B down for a while then the repeat function of the terminal sends several Control Bs to MicroBeta Note that Control B and Control N move only inside the default or the current value displayed before gt not inside a newly typed text The cursor control keys left and right arrows can sometimes be used instead of Control B and Control N This does not work with every terminal emulator The up and down arrow keys can be used when moving up and down in the protocol editor or you can type Lx to jump to line number x Please note that the Enter key must be pressed if making changes to any row As an example of keyboard editing suppose you have entered the following printout selection the second line Printer Output POS CTIME COCPMI CCPMIES gt POS CTIMR COUNTS CCPML CeEPML After you press Enter you will see the text
161. he counter control and press Enter to start counting Live data can be displayed by pressing F4 LIVE The results of each sample are displayed in plate format if plate coding has been used The results are then stored in MultiCalc for further evaluations see Stored files and Result files in the MultiCalc manual 2 22 5 6 Communication protocol See MultiCalc User Manual Communication protocols To change the RS232C port change only lines from 02 COMM PORT to 07 COMM MODE 101 2 22 MultiCalc operation 2 22 5 7 Communication problems See chapter 2 31 4 11 Terminal emulation GenTerm Trouble shooting for information about how to solve possible communication problems 2 22 5 8 Printer connection MultiCalc prints results to the printer connected to the terminal PC with a standard Centronics cable In terminal mode MicroBeta controlled assays direct printing also to the same printer 2 22 5 9 Exit to MS DOS In the main menu select first LEVELS F7 then MS DOS F8 See the MultiCalc User Manual for details of other menu operations 102 2 23 Normalization 2 23 Normalization See chapter 2J 23 for JET information 2 23 1 What is normalization MicroBeta has 1 2 3 6 or 12 detectors allowing it to count the respective number of samples simultaneously In order for the results from each detector to be equivalent irrespective of small variations in efficiency and background between detectors it is necessary to determine the
162. he current value is usually displayed before an arrow sign gt After typing the new value you must press Enter Press the Enter key alone to leave the default or current value unchanged To get help about the subject under question press Enter after H h or Press slash or backslash to exit input mode In the protocol editor exits with saving the protocol and without saving it Elsewhere usually both cause exit without saving the data Input data can be edited see below 2 1 4 Editing input data If you make a mistake when entering a new input value you can use the Backspace key sometimes labelled as BS or a thick left arrow or the DEL key to erase the previous character If the input data is quite long this can be tedious For this kind of situation a simple keyboard editing facility is provided Keyboard editing only uses a few keys so it is easy to learn The default value or the current value of input data is used as a template for entering a new value So if you notice that you have made a mistake during an early phase of entering a long input such as output selection in 16 2 1 Beginning operation of MicroBeta TriLux the protocol editor you can press Enter to complete entering a new value and then go back to the same line to correct the mistake The editing keys are lt or Control B Go back one character in the template default or current value gt or Control N Go to the next chara
163. he directory Cc UTERM into the MS DOS PATH variable 4 Copy the files START BAT TASK1 BAT and BPNAME EXE from the directory C UTERM into one of the directories listed in the MS DOS PATH variable Check that COMMAND com exists in the root directory C of the terminal PC hard disk 165 2 31 Terminal emulators 2 31 5 4 Starting UltroTerm a If installation was made on diskette either insert the terminal diskette into drive A and start up the PC or type START at the MS DOS prompt and press Enter b If installation was made on hard disk then you have three possibilities to start UltroTerm depending on the type of installation Start up the terminal PC installation d 1 Type CD C UTERM and press Enter then type START and press Enter installation d 2 Type START and press Enter installation d 3 and 4 2 31 5 5 Changing parameters in UltroTerm The main reason for communication problems is that the counter is configured to use one emulation and the terminal emulator uses some other emulation see 2 31 5 11 Troubleshooting below If there still seems to be communication problems after changing to the same emulation in both the counter and the PC the reason can be that there are different communication parameters in the counter and the emulator Here is a quick guide for how to change the communication parameters in UltroTerm For further information see the UltroTerm 2 User Manual 1 Press the E
164. he operation under question can affect the functioning of the instrument counting results etc Press Y Yes to accept the operation or N No to cancel it 2J 1 3 Input of data The rest of the user interface consists mostly of data input Data input differs somewhat from menu selection and confirming questions The default or the current value is usually displayed before an arrow sign gt After typing the new value you must press Enter Press the Enter key alone to leave the default or current value unchanged Either letter H h or character provide help text about the subject under question Press Enter after H h or Press slash or backslash Error Bookmark not defined to exit input mode In the protocol editor exits with saving the protocol and without saving it Elsewhere usually both cause exit without saving the data Input data can be edited see below 2J 1 4 Editing input data If you make a mistake when entering a new input value you can use the Backspace key sometimes labelled as BS or a thick left arrow or the DEL key to erase the previous character If the input data is quite long this can be tedious For this kind of situation a simple keyboard editing facility is provided Keyboard editing only uses a few keys so it is easy to learn The default value or the current value of input data is used as a template for entering a new value So if you notice that you have made a mistake during an early p
165. ials for blank 1 e crosstalk samples are needed b Add into each standard sample vial numbered from 1 to 6 scintillation liquid and isotope solution or pipette the isotope solution directly into the sample wells the same amount in each Add the same solution into blank vials wells but without the isotope If there is no isotope in the standard vials the same vials can be used for blank samples c Add quencher into each standard and blank vial so that the samples form a quench series 1 e the amount of quencher increases from vials 1 to 6 The amount of quencher added here CCl is used as a quencher forms a series as follows Vial No Amount of CCL 5 ml ul 0 5 15 30 50 75 DU Eu Dn kn Shake the vials Then pipette the standards and blanks onto a sample plate according to the following array use the same total volume in standards and blanks as in the actual samples 44 2 Crosstalk correction TATUJI W gt The isotope standard samples are marked with S and a number which refers to the vial number the amount of quencher The blank samples are marked with C and a number Close the sample plate with a sealing tape and place it on a 1450 103 counting cassette 2 7 3 2 Preparation of DPM standardization samples on a 24 well microtitration plate The procedure is similar to that described above but two plates are used and the sample positions are as follows First plate l 2 3 4 5 6 A cr O c3
166. icroBeta sends an identity inquiry to the terminal and the answer sent by the terminal emulator software is different for each emulation 154 2 30 System GenTerm UltroTerm VT52 VT100 Answer N if you want to cancel the new terminal emulation However if you know that your terminal has the correct emulation or you want to switch to the new emulation later you can answer Y Example 11 system gt M Terminal emulation 1 GenTerm 2 UltroTerm 3 MultiCalc A VT52 5 VT100 6 MBW 25 gt Number of lines on screen 24 gt Block graphics Y N Y gt Send results file to terminal Y N N gt Automatic result tile deletion Y N Y gt Print through terminal Y N N gt Do you accept new terminal setting Y N gt Prepare your terminal for emulation change Press any key when ready Terminal type check failed Do you accept the new terminal setting Y N gt N 2 30 11 P a sswords Press A to print protocol passwords entered on line 45 in the protocol editor see chapter 2 25 Protocols This is possible only if no System password is in use or in Setup mode see 2 30 15 Example 12 system gt A Printing protocol passwords press to exit 2 30 12 Customi z ing Press Z to set environment strings to customize the program The environment string name and value must be given For a list of possible strings see I nfo Customi z ing Giving D as the value sets the default value for the string Example 13 system gt Z Cus
167. ield of the cassette Select PLATEV3 in the counter control and press Enter to start counting NOTE The results files from the normalizations and standardizations will not be evaluated by MultiCalc 2 22 5 5 Counting unknown samples Unknown samples can be counted by using MicroBeta controlled protocols Counting protocols or MultiCalc controlled protocols Assay protocols The type of protocol is selected by putting an ID label with protocol number in the PROT field of the cassette for Counting protocol assays When using Assay protocols the ID label is attached to the ASSAY field The counting of samples using Counting protocols is started as when using MicroBeta without MultiCalc 1 e select Automatic counting in the Ready state It can also be started from MultiCalc by pressing Fl F COUNTER and then selecting PLATEV3 and pressing Enter The live data and results are displayed on screen and results are sent to each output medium as selected in the protocol Printer outputs are sent to the printer connected to the PC if Print through terminal is selected in System Ter m inal Results to file are sent through MultiCalc to the appropriate disk drive on the PC NOTE The results from assays using counting protocols will not be evaluated by MultiCalc The counting of samples using Assay protocols may only be started from MultiCalc Check that the protocol number label is in the ASSAY field of the first cassette in a batch Select PLATEV3 in t
168. ilable in the protocol are Chemiluminescence correction Half life correction Password protection 3 4 14 4 MicroBeta JET injector system One injector module dispensing at a time Reading before dispensing reading after dispensing and delay time after dispensing 3 4 15 Consumables 3 4 15 1 Sample Plates 1450 401 96 well Sample Plate 25 box 96 round bottomed wells 8x12 format made of clear PET plastic printed lines between wells chemically resistant to all HiSafe cocktails max volume 250 uL well Sealed with tape type 1450 461 or 1450 462 or heatsealed with 1450 463 Gin 1495 021 Microsealer Counted in cassette type 1450 101 1450 402 24 well Sample Plate 25 box 24 flat bottomed wells 4x6 format made of clear PET plastic printed lines between wells chemically resistant to all HiSafe cocktails max volume 1 mL well Sealed with tape type 1450 461 or 1450 462 Counted in cassette type 1450 102 1450 405 96 well Sterile Sample Plate 10 box 96 flat bottomed wells 8x12 format made of polystyrene white well walls and clear well bottom sterile tissue culture grade with a lid max volume 350 uL well For tissue culture samples and for samples with Polystyrene Safe cocktail 1450 444 Sealed with tape type 1450 461 or 1450 462 Counted in cassette type 1450 105 272 3 4 Specifications 1450 407 96 well Sample Plate PS 400 uL 25 box 96 flat bottomed wells 8x12 format made of clear polystyrene max volum
169. ill be selectable via softkeys You can now either select a factory made counting protocol by pressing an isotope softkey or select a protocol from the list Scroll the list by the up and down arrow keys or PageUp PageDown for faster scrolling and make the selection by pressing Enter Note If you have a newer version of MultiCalc but do not get the counting protocol list try to install the counter again Press F5 INSTALL in the counter mode and the counter will send the protocol index to MultiCalc See also the Introduction to MultiCalc operation manual the module on Protocol operations 99 2 22 MultiCalc operation 2 22 5 2 Plate coding Plate coding is accessed by pressing Fl CODING See the MultiCalc User Manual Plate operation Plate coding requires the replicate number and the sample type to be set Press F7 to increase or F8 to decrease the replicate number Press Fl to F4 to select the sample type Then press Enter to get a new line for the next replicate type The PLATE key F6 allows a sample layout to be defined directly onto the plate After pressing F6 a plate is displayed showing the different codes in different wells Use the cursor keys to move around the plate Select EDIT F1 to edit existing layouts or CREATE F2 to define a layout of your own See the MultiCalc manual for how to make the plate layout Press F9 to exit the plate coding Again press F9 to exit from protocol editor Select Fl to save ch
170. in This procedure continues for as many times as you have defined for the cycle parameter 2 6 8 Delay between plates This parameter can be used to control the interval between when the counting of one sample plate has finished and the next is started The range is O to 9999 minutes 39 2 6 Counting control 40 2 Crosstalk correction 2 Crosstalk correction 2 7 1 Introduction Crosstalk is a situation in which light pulses from adjacent samples interfere with the pulses of the sample under measurement This may occur in the case of microtitration plates in which you cannot use the cassettes 1450 101 or 1450 102 This is because designs of cassette other than the 1450 101 or 1450 102 have part of the optical shielding between the wells removed to accommodate the microtitration plate Crosstalk can be corrected with the MicroBeta program Before counting the actual samples the amount and type of crosstalk is defined using special standardization or normalization samples with the type of solution as similar to the actual samples as possible The normalization or standardization is done first and the calculated crosstalk factors are then used when counting the actual samples A CPM normalization run 1s needed for crosstalk corrected CPM results and a DPM standardization run is needed for crosstalk corrected DPM results 2 2 Crosstalk CPM normalization 2 2 1 Preparation of normalization samples on a 96 or 384 well microtitrati
171. in chapter 2 22 press M 111 2 25 Protocols To display the protocol list in the Protocols menu press S Show You will see a two column list of protocols containing protocol number name if given and isotopes The list of the counting protocols also contains counting mode CPM or DPM and the number for the normalization or standardization protocol used The lists of the normalization and standardization protocols have a column containing the date when the normalization or standardization has been done An equals character indicates that crosstalk correction is used If there are more protocols than there 1s space for on the display press Enter to get the next page of protocols When you have found the one you want to look at type the number of that protocol and press Enter Type L List to printout the protocol list and P Print followed by protocol number and Enter to print a particular protocol 2 25 3 Creating and editing protocols To create a new protocol with default values press E Edit Choose a protocol number not in the list and press Enter The first free protocol number is shown in the menu Protocols have already some values in them either default or old parameters If you want to set default parameters press F deFault and select the protocol number You will be asked for a line number when starting editing a protocol Press Enter to start editing from the beginning of the protocol or type a line number follo
172. is then defined to be one and the other detector efficiencies are expressed as a fraction of this value These fractions are called efficiency coefficients If isotope activity is given then absolute efficiencies are calculated by dividing the count rates by the activity In this way sample quench and detector absolute efficiencies can be corrected for When measuring a sample with a particular detector the CPM of the sample is corrected by dividing the CPM by the efficiency coefficient In dual label counting corrections are made using dual label correction formulae see 3 3 Calculation methods When normalization is done the results are stored with the normalization protocol The normalization data can be used by one or several protocols and is selected when editing the protocol see 2J 25 Protocols Normalization with injection The principle is the same as normalization without injection but there is a difference in the arrangement of the background and standards In addition parameter line 5 must be 9 Luminescence With other selections the Mask Adapter can be selected but not injection Instead of a background plate a background sample is used for each detector The position of the background sample and standards is shown in the plate maps in section 2J 23 4 2J 23 2 When is normalization necessary Each counting or assay protocol requires normalization data to be stored before it can calculate corrected CPMs for the samples cou
173. ite EEE hee eet cas aa tame uh Na hea E E E 215 2J 27 RODOUC loadin Interlace ii Rs 231 Be A 0 yg E EP II 233 Part 3 Instrument description Sl Sto Mentales CAPO ds a ds A 249 3 2 Roumie Munene E A daa 253 De AIC UAL ON MENOS een a N A 299 OAE CLIC all ONG o E E ET E acido E EE ET 265 3 9 Abbreviations and ACTON YMS US di A AA A 271 Declaration of conformity for CE marking eeeesesssseoeeessssssssecerrsssssssceresssssseeerresssssseeeeres 283 Part 4 Installation information AF MLNS Tel MAIO MTS CEU CUO INS iaa anio 287 4 2 Installation of the Injector System to MicroBeta JET ccccccnnononnnncnnnnnnnnnnnnncnnnnnnnnnnnnos 301 Part 5 Index A O 309 Trademarks Trademarks MicroBeta and MultiCalc are registered trademarks and Betaplate RiaCalc OptiPhase and Wallac are trademarks of PerkinElmer Inc IBM IBM PC AT IBM PC XT and PS 2 CGA EGA MCGA VGA and PC DOS are trademarks or registered trademarks of International Business Machines Corporation MS DOS and Microsoft are registered trademarks of Microsoft Corporation Amersham is a trademark of Amersham International plc Olivetti 1s a trademark of Ing C Olivetti amp C S p A Epson is a registered trademark of Epson Corporation VT is a trademark of Digital Equipment Corporation Intel is a registered trademark of Intel Corporation Macintosh is a registered trademark and System 7 and MultiFinder are trademarks of Apple Computer Inc MultiScreen is a trade
174. ith tape type 1450 461 or 1450 462 Counted in cassette type 1450 105 3 4 15 2 Filtermats 1450 421 Printed Filtermat A 100 box 96 position glassfibre filter mat with printed pattern For harvested cell and receptor samples Scintillant to be used Betaplate Scint 1205 440 or MeltiLex B HS 1450 442 Placed in a sample bag type 1450 432 Counted in cassette type 1450 104 1450 422 Filtermat A 100 box 24 position glass fibre filter mat with printed pattern For harvested cell and receptor samples Scintillant to be used Betaplate Scint 1205 440 or MeltiLex A 1450 441 Place in a sample bag 1450 432 Counted in cassette type 1450 116 1450 423 Printed Nylon Membrane 50 box 96 position nylon membrane with printed pattern for DNA samples Scintillant to be used Betaplate Scint 1205 440 or MeltiLex A 1450 441 Placed in sample bag type 1450 432 Counted in cassette type 1450 104 273 3 4 Specifications 1450 424 Filtermat B 50 box 24 position filtermat with printed pattern made of glassfibre For harvested or pipetted samples Scintillant to be used Betaplate Scint 1205 440 or MeltiLex B HS 1450 442 Place in a sample bag 1450 432 Counted in cassette type 1450 116 1450 521 Printed Filtermat B 50 box 96 position glassfibre filter mat with printed pattern For harvested receptor or cell samples Scintillant to be used Betaplate Scint 1205 440 or MeltiLex A 1450 441 Placed in sample bag type 1450 432 Counted in cassette ty
175. ities 100 250 or 500 uL Select the one you want 2J 30 14 9 Aspirating tube 1 D The internal diameter of the tubing from the liquid reservoir to the syringe The units are inches 2J 30 14 10 Aspirating tube length The length of the tubing from the liquid reservoir to the syringe The units are inches 2J 30 14 11 Dispensing tube I D The internal diameter of the tubing from the syringe passing through the injector assembly to above the well 2J 30 14 12 Dispensing offset This is the offset volume for injector modules and it is set in steps of 1 uL It is a volume that effects the injector dispensing volume If the volume is selected from the protocol as 10 uL and the offset is O then the dispensed volume is 10 uL If the offset is 2 uL then when the protocol selection is 10 uL the real dispensed volume is 12 uL So if in the injector there is for some reason a systematic error in the dispensing volume it can be corrected with the offset 244 2J 30 System The following parameters are common for all channels and should not be changed by the user without contact with a service person 2J 30 14 13 Dispense fast command string This defines how fast the dispensing occurs for fast dispensing 2J 30 14 14 Dispense slow command string This defines how fast the dispensing occurs for slow dispensing 2J 30 14 15 Dispense custom command string This defines how fast the dispensing occurs for the custom dispensing This can be set
176. ization data If automatic counting is selected the normalization cassette can be placed anywhere in the rack The NORM label is needed in this case The counting results for the normalization are output as specified on lines 30 37 in the normalization protocol and after them the background and detector efficiency values are given Note Normalization and unknown samples should have the same quench level in dual label counting 2 23 6 Preset normalizations There are four preset normalizations available in MicroBeta Normalization protocol 1 contains a H normalization using ScintiStrip plate 1450 419 dry 200 uL volume with crosstalk correction Normalization protocol 2 is the same as but using al Normalization protocol 3 contains a generally used H normalization Normalization protocol 4 contains a generally used 14C normalization 107 2 23 Normalization 108 2 24 P 32 Dot blot quantification 2 24 P 32 Dot blot quantification Assays utilizing the P label and 96 format filtermats e g dot blot hybridizations can be detected with MicroBeta using the 2P cassette 1450 118 Dried sample membranes can be counted directly in the cassette while wet membranes need to be counted in a sample bag to avoid any contamination The counting method does not include any scintillator addition to the actual filter and thus allows re probing of the samples When P samples are measured with the 1450 118 cassette the obtaine
177. izing The program can be customized using various environment strings These strings and their effects are listed under this control letter 2 15 10 S y stem This is a list of system information and settings 74 2 16 Interrupt 2 16 Interrupt 2 16 1 Stop If it is necessary to stop counting quickly press O Off The cassette currently in the counting position will be moved back to the rack and the program will enter the Ready state Note If you have started counting from MultiCalc and are in MultiCalc mode you cannot use the O key Instead you must press Fl STOP or if the softkeys are not in use then press any other key e g the space bar to stop counting 2 16 2 Open the door If the door is opened during counting the high voltage is cut off and the counting stopped An error message is displayed on the screen When the door is closed the program starts counting the current positions from the beginning 2 16 3 Next position or assay If you want to stop the counting of the current samples press key N Next position or E Next assay Counting will be interrupted and the MicroBeta will start to count the next position if N is pressed or the next assay if E is pressed If quick view is used and N Next plate is pressed then MicroBeta starts counting the next plate Similar keys in MultiCalc mode are F2 NEXT POS and F3 NEXT ASSAY 2 16 4 Causing a pause in printing It is possible to cause a pause in printing
178. l 12 det If Easy DPM is selected then only normalization standards are needed 66 2 12 DPM standardization If ParaLux counting is selected then it is possible to subtract thermal background from the AQP and counts values Select Background sample on line 42 and put an empty cassette as the first standardization cassette followed by the usual standardization cassette LinS la dd ENDS COto E Tor Telp 1 Protocol name gt 2 Sample type 1 Normal 2 SPA 32 Crosstalk Correct ron Y N 4 Number of labels 1 Single 2 Dual 5 Isotope 1 IYH 3 2TH ko o Gala 4 S 35 SIOL 6 PSs37 7 P 32 Cerenkov 8 Other 9 Luminescence ParaLux Y N ParaLux mode 1 High efficiency 2 Low background 26 30 32 34 36 40 41 42 43 45 46 47 Discriminator channel 1 PMT use 1 Normal 2 Upper 3 Lower Window Ls o gt 3S Counting time for crosstalk s 60 Precision 2 sigma Qe Easy DPM Y N Number of standards Isotope 1 activity DPM 200000 Standard curve fit selection Y N Curve fit method 1 Smoothing spline 2 Interpolation spline 3 Linear interpolation 4 Linear regression Automatic smoothing Y N Faust standaro CUEVe Y N Printer output 1 No 2 Snore 3 Long 4 Programmable Display Output LINO A MOMO 3 Long 4 Programmable External output 1 No 2 SHOLrt 3 Long 4 Programmable File output 1 No ZV Snore 3 Long 4 Programmable Change special features Y N Isotope activity setting Y N Background c
179. l be counted For a more detailed specification 1 e giving the positions instead of rows only press R Row coding In this case specify which positions are to be counted for each row The positions are in order from 1 to 12 or 6 for 24 well plates Type 1 to count a position or O to skip it E g Row A gt 0011110 means that positions A3 A4 AS and A6 should be counted You do not have to enter all the zeros at the end of the line the row end 1s filled with O or 1 according to the last typed character See the following example 20 Count all positions in cassette Y N Y gt N 21 Cassette type 1 96 2 24 i gt LL Cassette 1 96 Rows AB gt R Row AS TILLUETVTILELITL SS 0 00 TL T0 Row B 0071 100 0000 gt 0 000110 Row Cf 000011000000 gt 0 Row D 000000000000 Cassette 2 96 Rows ABCDEFGH gt AB Cassette 3 96 Rows ABCDEFGH gt An alternative method is to type the column numbers e g 5 6 7 equals 000011100000 1 equals 100000000000 and 3 equals 001111111111 After pressing Enter you will be asked for positions for the next row The default values are the same as you have given on the previous row Press when you want to stop specifying counting positions The rest of the rows will then be counted as the last edited row In the example above the following positions will be counted for cassette 1 A3 A4 A5 A6 B5 and B6 The following cassettes can then be coded in the same way as the first one Press once ag
180. l error of ISEP COUNTSy Counts for isotope z or extra window 3 CPMy Count rate counts per minute for isotope z or extra window 3 CPMy Percentage theoretical error of CPMy 1 CCPMy Corrected count rate for isotope z or extra window 3 11 CCPMyS Percentage theoretical error of CCPMy 1 DCCPMy Absolute theoretical error of CCPMy 1 127 2 26 Results DPMz DPMz DDPMz CLM EFFy STM DPMM FLAG Wy CESY CEPSy RESP 1 RESPZ BESPZS LCPS LCPS DECES LM AQP 1 AQP 1 CEM 6 CEM NCE CPM_AQP DPM of isotope z ii Percentage theoretical error of DPMz 1 Absolute theoretical error of DPMz 1 Chemiluminescence in window 1 Isotope z or extra window 3 counting efficiency Stats trics monitor LT DPM out of range monitor 111 Termination flag 111 Window y limits Count rate counts per second for isotope z or extra window 3 Corr count rate counts per sec for isotope z or extra window 3 ii CEDML sor CPS CEBMZ Percentage theoretical error of RESPz Luminescence corrected counts per second divided by 100 Percentage theoretical error of LCPS 1 Absolute theoretical error for LCPS 1 LEPS out OL range monitor LLL Asymmetric quench parameter 0 0 1024 0 Percentage theoretical error of asymmetric quench parameter Coincidence CPM1 Non coincidence High efficiency CPM1 CPM_C CPM_NC CPM used in AQP I calculation c Statistical data STAT Sil REPL MEANZ
181. l the sample cassettes in the rack can be counted in repeating cycles One cycle means counting of all cassettes in a batch once i e those being counted with one protocol A batch ends when the next cassette with a protocol ID is found When the instrument has counted all the cassettes in this batch it moves the rack up until the first cassette in the batch is found Then it recounts the cassettes in the batch Counting of the next batch starts when the cassettes have been counted the number of times specified by the Cycle parameter The repeating cycles can be used to check sample stability The maximum number of cycles are 99 If the number of cycles is greater than 1 then you will be asked to give the Cycle delay This is in minutes and is the time between when one cycle ends and the next one begins 2 6 7 Order of operation of repeated counting If you have set two or all three of the repeat replicate and cycle parameters to values greater than 1 the order of operation 1s First repeat counting of each sample The output consists of the individual results for each repeat count for a sample followed by statistics for the repeat Second statistics for all the replicates of the sample are calculated using the statistics for the repeat 38 2 6 Counting control This procedure is repeated for all samples in an assay batch When the end of the batch is reached counting starts from the beginning by counting the repeats and replicates aga
182. late orientation 1 Normal 2 Rotated 3 Mirrored 4 Both als 30 Printer output 1 No 2 SHOrC 3 Long 4 Programmable 3 32 Display OMEDBUES LINO ZO MO 3 Long 4 Programmable 3 34 External output 1 No 2 Short 32 Ong 4 Programmable 36 Fileoutput 1 No 2 NS NOTE 3 Long 4 Programmable I 40 Change special features Y N Y 41 Counting control YN Y Number of repeats a Number of replicates il Number of cycles 1 42 Background correction Y N Y Use normalization background Y N N Background sample Y N X Bgnd counting time s 1 0 43 Half life correction Y N N 45 Use password Y N N 48 Delay between plates m 0 217 2J 25 Protocols 2J 25 5 Protocol copy and delete The contents of one protocol can be copied to another by pressing C Copy Select the No of the protocol to be copied and enter the new protocol number If you want to delete a protocol press D Delete and select the protocol number Press Q Quit to return to the Ready state 2J 25 6 Protecting protocols Protocols can be protected against unauthorized editing by using passwords The password is selected on line 45 in the protocol editor This prevents other users from changing or deleting your protocol but they can still use it e g listing copying and counting with it The MicroBeta supervisor has the authority to list the passwords 2J 25 7 Parameters in a counting protocol 1 Protocol name Type a name for the protocol maximum 170 characters
183. lation c If installation was made on hard disk then select the way how GenTerm should be started as follows these instructions suppose that GenTerm was installed into directory C GENTERM 1 If GenTerm should start automatically after terminal PC start up copy lines of C GENTERM GTERM BAT to the file Cc AUTOEXEC BAT e g using the COPY command COPY C AUTOEXEC BAT C GENTERM GTERM BAT C AUTOEXEC BAT 2 Execute GenTerm always from the directory C GENTERM 3 Insert the directory C GENTERM to the MS DOS PATH variable 4 Copy file c GENTERM GTERM BAT into one of the directories listed in the MS DOS PATH variable d In the GenTerm setup menu set the Buffer length to a minimum of 100 characters 2 31 4 4 Starting GenTerm a If installation was made on diskette either insert terminal diskette into drive A and start up the PC 161 2 31 Terminal emulators or type GENTERM at the MS DOS prompt and press Enter b If installation was made on hard disk then you have three possibilities to start GenTerm depending on the type of installation Start up the terminal PC installation c 1 Type CD C GENTERM and press Enter then type GTERM and press Enter installation c 2 Type GTERM and press Enter installation c 3 and 4 2 31 4 5 Changing parameters in GenTerm The main reason for communication problems is that the counter is configured to use one emulation and the terminal emulator uses som
184. left to right and top to bottom 3 and 6 det models without background samples Rows C D are empty in the 3 det model 204 2J 18 Luminescence counting l and 2 det models without background samples Row C is empty in the 1 det model B C 28 SSS Bo A d IESO l and 2 det models with background samples Row C is empty in the 1 det model 2J 18 6 Normalization protocol Edit the normalization protocol so that it is suitable for luminescence measurements To do this in the Ready state press P P rotocol then N N ormalization protocols Select edit E and the protocol number Edit the protocol parameters See chapters 2 23 and 2J 23 for examples of normalization parameters Select luminescence 9 on line 5 isotope 1 and the counting time on line 10 Typically this is 1 s If required you should also select the mask adapter and if you want injection then the number of injector modules must be given 1 e not zero These are both part of the settings for parameter 5 For glow type luminescence a background plate is counted only if selected on line 42 For flash type luminescence using injectors a background plate is not used but instead if required background samples as described in sections 2J 18 4 and 5 above Make sure the injectors are ready for operation as described in chapter 2J 0 Injector operation 2J 18 7 Normalization counting First place the empty background plate in a cassette if you are using s
185. lizations begin with N e g N1 001 and standardizations with an S e g S1 001 The disk drive A B for floppy disks C and D for hard disks F etc for LAN drives and the path e g MB DATA for the result files are chosen after the File output This over rides any path set in System mode The drive and path defined in the System state 1s the default value see chapter 2 30 9 System Data drive This is the terminal PC path If results should be stored on the MicroBeta disk then the path must begin with an O e g B DATA If a VT52 or VT100 terminal is used then results are stored on the protocol disk as default directory B RESULTS see 2 30 10 Terminal The file name extension the part after the period can be changed on the next line This must be a number in the range 1 to 999 Note this line is not available if UltroTerm is used 120 2 25 Protocols 40 Change special features Press Y to change special features These are counting control background half life and chemiluminescence correction password and special plate type 41 Counting control This and the following questions will be displayed only if Yes has been selected on the previous line Press Y to change the counting control parameters These are number of repeats replicates and cycles If N is selected then these parameters have no effect The number of repeats is the number of times each sample will be counted The minimum value is
186. lize command string 245 Injector 242 Counting protocol parameters 306 Initializing 306 Installation 301 Priming 306 Test volumes and alignment 306 Injector channels 183 Injector module Parameter setting 208 Injector 1 244 Injector assembly 251 266 Injector module 183 201 243 251 Injector setup 183 Clear conveyor 186 Dispense 185 Dispenser IN OUT 185 Goto pos 186 Init 184 Next pos 186 Off 186 Prime 184 Status 186 Suckback 185 Tip 184 Wash 185 Input file 51 Inserts 275 Installation 287 Instrument description 249 Interrupt 75 Inverted plate 117 222 ISEP Isotope spectrum end point 258 Isotope 115 219 Activity setting norm 124 228 Activity setting stand 125 229 Spectrum end point 258 Isotope 2 activity 125 229 J Jammed transportation system Loading cassettes 78 312 L L 69 Labels number of 115 218 LAN Local area network 151 237 LCPS out of range monitor 93 Lid 75 Loading cassettes 77 Local area network 151 237 Long output mode 135 Low background 115 219 Luminescence Coloured sealing tape 201 251 Counting 79 201 Flash 201 Flash normalization 202 Glow 201 Glow normalization 201 Normalization 79 PMT selection 115 219 Units 201 Luminescence assays Dual luminescence 251 Flash 251 Glow 250 Kinetic measurements 251 M M 59 Mains voltage 289 Mask adapter 185 243 251 253 266 Robotic loading interface 231
187. llation 296 MultiCalc 96 Terminal emulators 159 Exit 160 Startup 160 Temporary exit 160 Terminal 159 What they are 159 Terminal operation 33 Terminal PC 49 Port 1 49 Terminal settings exiting 154 240 Termination flag 94 130 Test function 294 Test level 294 Thermal background 106 210 Time 27 Counting time 28 Elapsed time 28 End time 28 Tip 184 Tip init command string 245 Total count rate 177 Transportation system 249 266 U UltroTerm 152 165 238 Changing parameters 166 315 5 Index Exit to MS DOS 167 Installation 165 Printer connection 167 Restrictions 167 Result files 167 Special features 167 Start up with 290 Start up 166 Trouble shooting 167 UltroTerm start up Start up 291 Usage 74 Use injector 219 Use mask adapter 219 UTMAC 171 Disk full 172 Exit 172 Installation 171 Parameter changing 171 Printer connection 172 Result files 172 Special features 172 Start up 171 Troubleshooting 172 y Version 148 234 VT terminal start up Start up 291 VT100 153 239 VT52 169 VT52 96 153 169 239 Installation 169 Printer connection 170 Restrictions 169 Result files 169 Special features 169 Troubleshooting 170 W Wash 185 Wash command string 245 Window 116 179 219 Settings 179 Wrong key pressed 69 5 Index Z Zero date Half life 71 Z Zero time DPM standardization 65 Half life 72 316
188. lour quench kits for use with MicroBeta These are 89 2 20 Micro volume LSC H SPA PVT colour quench and calibration kit code TRKQ 7080 123 SPA PVT colour quench kit code RPAQ 4030 Note for more details of DPM standardization procedure with MicroBeta see chapter 2 12 DPM Standardization Load the standardization plate into a cassette with a STD label in the FUNC position and a suitable standardization protocol number and load this standardization cassette into MicroBeta Edit the standardization protocol and set the standardization parameters shown overleaf In particular select the SPA mode mode 2 This automatically selects ParaLux counting Run the standardization The results will be saved under the standardization number marked on the cassette When you come to run the actual SPA set the counting protocol for DPM and give the standardization protocol number selected above Other parameters should be set according to the instructions in your SPA kit insert 2 Sample type 1 Normal 2 SPA a2 5 Isotope 1 ie ps oe ZALLA COEL AVIS SS Ser Sl 10 Pas 7 P 32 Cerenkov 8 Other SS e SST Or 2 ISZ ParaLux Y N SS Y ParaLux mode 1 High efficiency 2 Low background gt 2 Discriminator channel gt 150 PMT use 1 Normal 2 Upper 3 Lower gt Window 1 default gt 10 Counting time for crosstalk s gt 600 sec 20 Number of standards gt 10 21 Isotope 1 activity DEM gt x where x is the activity of a
189. low or a custom speed set in the system settings 219 2J 25 Protocols Dispensing modules here you give the number of modules to be used If you have a single detector instrument you can have up to four injector modules If you have 2 3 or 6 detectors you can have one or two modules In order specify the modules used use the following codes One module or only the first module 1 The second of two modules 10 Both of two modules 11 The third of three modules 100 The first and third of three 101 etc up to All four of four modules 1111 6 Isotope 2 Displayed only in dual label counting The higher energy isotope 1s selected The alternatives are the same as for parameter 5 with 14C as default isotope 7 Sample volume Displayed only if Use injector has been selected to be Y This parameter defines the original amount of sample in the well before dispensing This is a mandatory entry and it is used for checking that the dispensed volumes do not exceed the maximum allowed volume The value is an integer number in micro litres uL The minimum value is 0 The default value for this entry field is empty 8 Dispensing volume Displayed only if Use injector has been selected to be Y This parameter defines the amount of liquid to be dispensed The value is an integer number in microlitres uL The minimum value is 10 uL The default value is 10 uL There is a separate parameter for each module used In a single detector counter
190. lts DPM counting is therefore needed DPM counting involves the use of quench curves These are made after counting a number of standards with different quench levels This procedure is called DPM standardization The spectra from counting unknown samples are compared with the quench curve and accurate results are calculated In MicroBeta the quench curves are constructed by first counting 2 to 12 DPM standards in detector and then counting two so called DPM normalization standards in every detector The curve from detector is recalculated for the other detectors depending on the detector efficiencies See chapter 2 12 DPM standardization for more information about the standardization procedure 2 11 2 Single label DPM The DPM standards are counted in detector 1 MicroBeta has no external standard so the spectrum quench parameter for the isotope SQP I or the Asymmetric quench parameter for the isotope AQP I is therefore used for determining the quench curve AQP I is used when ParaLux counting is selected The normalization standards are then counted in every detector in order to get the efficiencies and SQP I or AQP I values Later when measuring an unknown sample the CPM and SQP I or AQP I values are counted in a particular detector SQP I or AQP I is modified to fit as SQP I or AQP for detector 1 The efficiency for detector 1 is counted from this SQP I or AQP I by using the quench curve and is then used for
191. lues will be sent to output The maximum number of replicates is 99 2J 6 6 Number of cycles All the sample cassettes in the rack can be counted in repeating cycles One cycle means counting of all cassettes in a batch once i e those being counted with one protocol A batch ends when the next cassette with a protocol ID is found When the instrument has counted all the cassettes in this batch it moves the rack up until the first cassette in the batch is found Then it recounts the cassettes in the batch Counting of the next batch starts when the cassettes have been counted the number of times specified by the Cycle parameter The repeating cycles can be used to check sample stability The maximum number of cycles are 99 If the number of cycles is greater than 1 then you will be asked to give the Cycle delay This is in minutes and is the time between when one cycle ends and the next one begins 2J 6 7 Order of operation of repeated counting Without using injection If you have set two or all three of the repeat replicate and cycle parameters to values greater than 1 the order of operation 1s First repeat counting of each sample The output consists of the individual results for each repeat count for a sample followed by statistics for the repeat Second statistics for all the replicates of the sample are calculated using the statistics for the repeat 198 2J 6 Counting control This procedure is repeated for all samples in an
192. m counting time is 999 999 9 s more than 11 days 2J 6 3 Precision 2 sigma The statistical uncertainty in samples is expressed as the standard deviation or sigma value and is calculated by dividing number one by the square root of the counting value E g 10000 counts gives a one sigma value of 1 The 2 sigma value is two times the one sigma value or 2 for 10000 counts In MicroBeta precision is given as 2 sigma in a range from 0 to 99 9 Zero means that no precision test is used Make sure that the counting time is set long enough if precision is wanted E g two samples of 1000 and 10000 CPMs will take 10 min and 1 min respectively to achieve a 2 sigma value of 2 Precision is checked once a second The termination flag in results output will be set to 197 2J 6 Counting control PREC if the precision has been reached The counting stops if the precision has been reached in all detectors 2J 6 4 Number of repeats To check sample stability or to check instrument performance the same sample can be counted repeatedly up to 999 times If the repeat value is less than or equal to 5 then results are sorted and some statistical values are calculated and printed after the sample results 2J 6 5 Number of replicates To evaluate sample preparation errors a number of replicates of a sample can be used The results for each sample are output separately After each group of replicate samples some statistical va
193. mark of Millipore Corporation Part 1 Introduction 1 Introduction 1 Introduction 1 1 Guide to the Instrument manual There are two versions of Wallac 1450 MicroBeta from PerkinElmer Life and Analytical Sciences described in this manual MicroBeta TriLux and MicroBeta JET Most of the information 1s common to both models where there is a difference it is noted In Part 2 of this manual alternative chapters with the number 2J xx are included to describe JET features The xx is the same number as the equivalent chapter for MicroBeta TriLux Normal start up is described in section 1 2 On the following pages you will find compressed operating guides for MicroBeta TriLux and MicroBeta JET Operations include references to the appropriate chapter in part 2 or part 2J of this manual where you will find a more detailed explanation If you are using MicroBeta workstation software refer to the separate User manual instead of section 2 in this manual You only need to refer to section 2 if you work in terminal mode Part 3 of the manual describes the instrument including specifications There is also chapter describing the routine maintenance to be done by the user Part 4 is only needed during installation Part 5 is the alphabetical index 1 Introduction 1 2 Instrument start up MieroB eta TiL start up Microbets JET ari i 1 Derin i Praec Pria j 1 Finer tor PE J tok de Stak ioe Px 3 Protocol Program lor wrk ada on lor
194. me Select Setup mode Set serial port parameters Set printer parameters Select drive for data saving results Set terminal parameters Print protocol passwords Set environment strings to customize the program Set thermostat optional Set instrument serial number Edit temperature calibration values Set detector usage during counting set program support for the ParaLux board and set program support for the plate ID reader if installed and specify 1 16 32 shelf model 196 2J 6 Counting control 2J 6 Counting control 2J 6 1 Counting parameters The counting process is controlled by the following parameters in the counting protocol 10 Counting time 12 Precision 2 sigma 4l Counting control Number of repeats Number of replicates Number of cycles Maximum channel counts a hidden parameter 48 Delay between plates Note Assay protocol parameters are described in chapter 2 26 MultiCalc A CPM normalization protocol or DPM standardization protocol does not include parameter 41 Counting control Instead the crosstalk normalization and crosstalk standardization protocols contain counting time for both crosstalk samples and standards e g 10 Counting time for crosstalk 11 Counting time for standards 2J 6 2 Counting time Counting will be stopped after the specified counting time in seconds has elapsed from the start of the counting unless some other conditions have already stopped it The maximu
195. me is the current time See also chapter 2 14 Half life 44 Chemiluminescence correction Press Y to use chemiluminescence correction otherwise N default This correction is done with a correction formula see chapter 3 3 Calculation methods 226 2J 25 Protocols 45 Password Press Y to select a password if you want to prevent other users from changing your protocol Enter the password maximum 12 characters This will NOT be echoed to the screen You will also be asked to retype it to prevent typing errors The protocol cannot later be edited or deleted without knowing the password You must therefore try to remember it Only the MicroBeta supervisor has the authority to list the passwords see chapter 2J 30 System Passwords He or she can help you if you have forgotten your password 46 Special plate Select Special plate type Wallac is the standard plate Betaplate filtermat plates and other 24 well plates have different dimensions and are counted using just one detector However it is possible to count other 24 well plates with all detectors This selection comes after the plate dimension questions For Betaplate filtermats you can select the 96 well plate format output This is a composite of four 24 well plate format outputs If you select the Other option for plate type you must give the plate dimensions These are the distances of the centres of the Al A6 and D1 wells from the plate sides Special cassettes must be used for
196. mn 130 32 gt Data file F USR GUEST IHME 33 003 Lo gt Counting Protocol not3o nu 2 May 2000 15 36 16 gt Name TEST L6 gt CPM amormalizat 06n protocol mo 40 8 gt Assay Prot 33 Cass Func Cassette mos 17 Shelrss 4 6 PlatelD 4 gt POS CLLME CCPM1 CCPMIs AOL DOE ovo L2 BOG 25 24 90972 1 2 1 gt Bot Bore 66020 129 BOG Dey TEL des Doe DT DOE 2 2 2 gt 8 gt Assay Prot Cass Func Cassette nos 27 Sheilt 4 FAG 4 gt POS ETTME CCPM1 CGPM Is AOL DO 8065 2 A06 29 7 90963 1 2 1 gt BOl 29 7 86122 1 2 Boe DOT 76035 Ws Boe Bo IB 3 16 gt Total count rate 4130593 2 CCPM ISSO 300 MELO OS KBE 32 gt End of assay 2 26 Results You will notice that certain lines are tagged with a number and an arrow In normal operation these line tags are hidden but they are shown here to help explain this function 131 2 26 Results The EXCL keyword can be used to mask out of the output whichever of the tagged lines you want left out To do this you must use the code EXCL followed by two numbers separated by colons The format is EXCL lt begin gt lt rest gt where lt begin gt sum of the numbers used to tag lines you want to exclude before and during the first cassette output add 64 if not greater than lt rest gt see below lt rest gt sum of the numbers used to tag lines you want to exclude during output from other cassettes and after the last cassette
197. moothing spline is selected If the DPM out of range monitor heading DPMM is selected then OUT 1s printed to warn the user The interpolated efficiency is corrected to correspond to detector 1 using the equation E EG N2 EG N1 EG N2 EC N1 E N2 EC EC N2 21 Activity o sigma percentage error headings DPM and DPM1 is given by a SA A a St r SE E 7 22 where or r is count rate error 6 and efficiency error comes from the equation SE V k 5q w 23 Here k is the slope of the standard curve and w is the standardization weight at q 3 3 Chemiluminescence correction Chemiluminescence correction is done by subtracting the chemiluminescence counts from window 1 c corr c c chem 24 Chemiluminescence counts are calculated using the equation c chem 7 4E 9 c l c r 25 where c l and c r are the lower and upper detector counts respectively The percentage of chemiluminescence in window 1 heading CLM is given by 1 100 c c corr c 26 260 3 3 Calculation methods If CPM1 lt 10 then chemiluminescence percentage is always zero 3 3 8 Statistics monitor If statistics monitoring is selected heading STM then the counting time is divided into ten parts and the samples are recounted if bad statistics is detected The counting time must be at least ten seconds for the statistics monitor to function E g 53 seconds 1s divided into 10 times 5 seconds and the last
198. n Press Y to use standard curve fitting otherwise N If No is selected smoothing spline with automatic smoothing will be used or linear interpolation if SPA is selected for the sample type 25 Curve fit method This line is displayed only if you have selected Yes on the previous line The methods for standard curve fitting are 1 Smoothing spline default 2 Interpolation spline 3 Linear interpolation default for SPA 4 Linear regression Select a method by typing the number and Enter If you have selected the first alternative smoothing spline you will be asked if the method is to include automatic smoothing Press Y 125 2 25 Protocols default if so Otherwise press N and you will be asked for a smoothing parameter Enter a value between 0 0 no smoothing and 1000 0 maximum smoothing with 1 0 as default 26 Edit standard curve Press Y to edit the quench parameter efficiency and weight values for a standard curve or to delete curve points 41 Adjusted activities Press Y if you want to adjust the activities for standards otherwise the activity given on line 21 will be used for every standard After pressing Y you will be able to change activities for the same number of DPM standards as given on line 20 You can also change the activities for the two Easy DPM standards The range is from 1000 0 to 999 999 9 DPM The value given on line 21 1s default See chapter 2 12 DPM Standardization for further information
199. n To get to terminal mode select Fl COUNTER from the MultiCalc main menu followed by F3 TERMINAL In this mode MicroBeta can be used as with any other terminal emulator Press F9 EXIT to exit from terminal mode and go back to the normal MultiCalc counter mode Press F10 HELP to get assistance in almost every situation 2 22 4 Assay protocols The main difference between operating MicroBeta with a terminal emulator or operating it with MultiCalc in counter mode is in terms of the data handling abilities In the former case everything is determined by the MicroBeta counting protocol which is what is described in most of the other chapters in part 2 of this Instrument Manual Counting protocols are normal liquid scintillation counting protocols containing parameters which control counting see chapter 2 25 Protocols Parameters in a counting protocol These are used e g edited without MultiCalc or in the terminal mode of MultiCalc Assays with only counting protocols can also be started from MultiCalc but the results are not evaluated by MultiCalc The PROT ID on the cassettes identifies assays using Counting protocols If however you are using MultiCalc counter mode then you work with MultiCalc assay protocols Assay protocols are connected to counting protocols An Assay protocol contains protocol number name counting time and precision plate coding and outputs Other parameters such as isotopes CPM DPM mode corrections etc
200. n device by connecting it to port 2 The external output and the result format are chosen on protocol line 34 External output 34 External output 1 No Z Shore 3 Long 4 Programmable 1 gt 4 35 External output POS GCCPM1 CCPM1 2P06 CTIME CPM CPM CCPM CCPM1 There are four alternatives which you can select on protocol line 34 External Output 1 Nothing is sent 2 Only position number CCPMs and CCPM DPM and DPM or LCPS LCPS and FLAG are sent CCPM is corrected CPM and means the percentage error in the CCPM DPM or LCPS value CCPM DPM or LCPS are also sent in PLATE format 3 The long results printout including SQP I and CPMs is sent see the Results chapter 4 User selected outputs are sent 2 8 6 Data processing Counting results are stored in ASCII format and they can be processed using commercial spreadsheet programs 2 8 Datafiles in MultiCalc Results from MultiCalc controlled Assay protocols are handled in a different way The types of results files are selected in the Assay protocol editor Input is an internal file for saving data to be used later in MultiCalc e g in evaluations Results is a text file to be used mainly for transferring data to an external computer See the User Guide to MultiCalc functions manual and the module called File handling and evaluation 51 2 8 Datafiles Results from MicroBeta controlled protocols are handled as with any other terminal emulator 1 e they are sent via
201. n dual label counting In DPM counting DPM1 and DPM1 are added In luminescence counting the output is PROT STIME POS CTIME CPS1 LCPS LCPS 222 2J 25 Protocols 4 Programmable The output items and arithmetic expressions are defined separately on the next line Sample quality monitors can also be selected in the programmable output Press or H to get a list of output items monitors and rules for using own formulae See also chapters 2 1 Beginning operation 2J 23 Results 2 29 Statistics and 2 21 Monitors and flags If SPECTRA is selected three more lines are shown The first asks for the energy scale to be selected It can be logarithmic with the units in channels or linear with the units in keV or both of these Next you are asked to specify the range of the plot This will be channels or keV depending on your previous selection Finally you are asked for the spectrum size small medium or large A smaller size will give faster output The two figures following illustrate the type of plots you may get POS C TIME CCPMi TEPMI AGS 58 7 2143516 1 O 4 Counts Seale 1 o 1060 200 200 aot ENS 400 TOG Channel Spectrum with logarithmic scale Spectrum with linear scale 223 2J 25 Protocols 32 Display output This output is selected when you want to display results on the screen It can be chosen independently of the printer output Display output has the same output alternatives as the printe
202. n the outputs For the terminal emulator the data coming from the RS 232C port looks as follows Display printer output lt open gt lt path gt xx yyy lt CR gt Open file xx yyy in directory lt path gt lt begin gt POS CTIME CCPM1 save this data into file xx yyy lt end gt lt begin gt A01 60 102356 save this data into file xx yyy lt end gt lt begin gt 402 60 TL52 94 save this data into file xx yyy lt end gt lt close gt close file xx yyy where xx protocol number yyy running result file number lt CR gt carriage return character and lt path gt drive and directory set in System D ata drive 2 01 6 5 Printer connection VT52 and VT100 terminal emulations have control codes to direct output to the printer connected to the terminal These codes are VT52 lt esc gt W Printer controller mode on lt esc gt X Printer controller mode off VT100 lt esc gt 51 Printer controller mode on lt esc gt 41 Printer controller mode off All characters between printer controller mode on and off codes will be transferred to the printer All other characters will be displayed However usually some control character NULL backspace etc not directed to the printer can disable graphics printing 2 31 6 6 Troubleshooting For communication problems see GenTerm Troubleshooting 1 170 2 31 Terminal emulators 2 31 7 UTMAC 2 31 7 1 Product number and manual 1221 170 UTMAC 1 1 Termin
203. nd hence different SQP I values The SQP value is then used to obtain the efficiency and hence the DPM for the sample In order to automate dispensing to a 96 well plate you can use a device such as the Labsystems Multidrop liquid dispenser 2 20 8 Scintillation Proximity Assay SPA 2 20 8 1 Introduction The scintillation proximity assay developed by Amersham is an application for which MicroBeta with the ParaLux counting feature see section 2 20 9 below is very well suited In SPA fluomicrospheres coated with receptors or acceptor molecules are used for binding the ligand to be assayed These fluomicrospheres also constitute the scintillation medium Low energy electrons e g tritium beta particles or I 125 Auger electrons from the radioactively labelled bound ligand cause light emission from the fluomicrospheres Labelled unbound ligand does not cause light emission because of the dilute concentrations used in bioassays where the distances between the unbound ligand and the fluomicrospheres is greater than the maximum range of the low energy electrons they are absorbed in the aqueous medium Consequently there is no need to separate the unbound ligand The whole assay is performed in the microtitration well and the result counted in a beta scintillation counter such as MicroBeta 2 20 8 2 Colour quenching and SPA If the SPA involves coloured solutions then there may be a need to make a colour quench correction Amersham provides co
204. ng control YN Y Number of repeats 3 Number of replicates 1 Number of cycles 1 42 Background correction Y N Y Use normalization background Y N N Background sample Y N Y Bgnd sample position G12 Bond Countang time s 60 43 Half life correction Y N Y Hali 1ife 1 If 10750000 Zero time 1 set Y N Y Aero date 1 5 Mar 2000 Zero time 1 10 18 46 Half life 2 h 0 00 44 Chemiluminescence correction Y N N 45 Use password Y N N 48 Delay between plates m 0 2 25 5 Protocol copy and delete The contents of one protocol can be copied to another by pressing C Copy Select the No of the protocol to be copied and enter the new protocol number If you want to delete a protocol press D Delete and select the protocol number Press Q Quit to return to the Ready state 2 25 6 Protecting protocols Protocols can be protected against unauthorized editing by using passwords The password is selected on line 45 in the protocol editor This prevents other users from changing or deleting your protocol but they can still use it e g listing copying and counting with it The MicroBeta supervisor has the authority to list the passwords 2 25 Parameters in a counting protocol 1 Protocol name Type a name for the protocol maximum 170 characters only the first 16 characters will be shown in the protocol list and press Enter The name may not start with characters or 114 2 25 Protocols 2 Counting mode Type
205. ng 1450 407 plates Cassette 1450 104 This is a two piece cassette with 96 sample holes This cassette is used when counting filtermats The filtermat in a sample bag is placed between the base plate and the cover plate Cassette 1450 105 This has 96 sample holes This cassette is used when counting 1450 410 1450 405 1450 419 or equivalent sample plates Cassette 1450 106 This has 96 sample holes This cassette 1s used when counting e g Millipore MultiScreen Filtration plates or equivalent sample plates 25 2 2 Cassettes Cassette 1450 107 This is a two piece cassette with 24 sample holes The cassette 1s used when counting Skatron filtermats cut into four pieces The filtermat in a sample bag is placed between the two plates of the cassette Cassette 1450 110 This is an open based cassette The cassette is used when counting 24 well culture plates e g Costar plates or equivalent Cassette 1450 116 This is a two piece cassette with 24 sample holes This cassette is used when counting filtermats The filtermat in a sample bag is placed between the base plate and the cover plate Cassette 1450 117 This is a two piece cassette with 24 sample holes This cassette is used when counting 4 mL sample vials The vials are placed between the base plate and the cover plate Cassette 1450 118 This is a two piece cassette with 96 sample holes This cassette is used when counting e g P labelled dot blot samples from filterma
206. nnels to be used You must also specify the volume of liquid to be used for dispensing The units are microlitres and the maximum value is 30 000 uL 185 2J 0 Injector setup 2J 0 12 Co m mand Pressing M allows injector specific macros to be sent to the injector unit This is mainly for service purposes 2J 0 13 S tatus If you want to see the status of the injectors then press S If there 1s an error condition you will be informed of it Otherwise you will be told that the system is ready for operation 2J 0 14 O f f Press F to switch off the injectors 2J 0 15 N ext pos When you have the mask adapter and a plate in position you can press N to move the plate to the next position ready for dispensing to the next well 2J 0 16 G oto pos Press G to move the plate to a selected well position 2J 0 17 C lear conveyor Press C to move the plate from under the mask adapter back to the cassette rack 186 2J 1 Beginning operation of MicroBeta JET 2J 1 Beginning operation of MicroBeta JET The following conventions are followed all through the MicroBeta user interface 2J 1 1 Menu selections Menu selections can be entered when the system is 1n one of the following states Ready gt Ready for next operation ProtocoLs gt Protocol Operations Coume gt Counting operations INLO Additional info System gt System level operations Counting protocol gt CPM normaliz Pprotocol gt DPM standard protocol gt Assa
207. nted A fresh normalization must be done when a new 207 2J 23 Normalization isotope is counted or when counting features such as isotope or window are changed When a new type of microtitration plate is used crosstalk normalization may be necessary 2J 23 3 Editing normalization parameters Normalization parameters are stored in normalization protocols These parameters are shown in the figure for the case in which injection is selected 1 e luminescence and mask adapter are both selected Note If you do not want injection then set the dispensing modules parameter to zero Otherwise set the number of dispensing injector modules using the code described on the next page Lane ikes 6 type to exit E Tor ne lp 1 gt 1 Protocol name gt it lash Juma norm Normalization not done 3 Crosstalk Correction Y N N gt 4 Number of labels 1 Single 2 Dual 1 gt 5 Lsotope 1 IS 77 EAS O 4 5 235 SICr 51 3 6 P 32 7 P 32 Cerenkov 8 Other 9 Luminescence 9 gt PMT use 1 Normal 2 Upper 3 Lower 2 gt Window 1 5 1024 Use mask adapter Y N N gt Y Dispensing 1 High 2 Low 3 Custom I e Dispensing modules LE QU 7 Sample volume uL 100 gt 8 Dispensing volume uL Gh gt 9 Delay time after dispensing s 1 0 gt 10 Counting time s 1 0 gt 12 Precision 2 sigma 0 2 gt 30 Printer output 1 No 2 Short 3 Long 4 Programmable 3 gt 32 Display SUEPpUES LINO Z LS NOE 3 Long 4 Programmable 3 gt
208. nter key Then follow the instructions displayed on the screen Now store the copied diskette as a backup copy and use the other diskette as a protocol diskette in MicroBeta or if a new empty protocol diskette was made use it for saving new protocols To get back to the MicroBeta program put the program diskette back into drive A type MB and press Enter 58 2 10 Diskettes 2 10 6 Preparing a 3 5 protocol diskette A standard 3 5 MS DOS 1 44 MB formatted diskette can be used as a MicroBeta protocol diskette When you want to initialize an empty formatted diskette to be used as a protocol diskette insert the empty diskette into drive B on the counter and start up MicroBeta When the program has been loaded the following message should appear Cannot find the protocol index Press M The program will try to find the protocols on the diskette and will create the protocol index Then the program returns to the Ready state and the protocol diskette is ready for use 59 2 10 Diskettes 60 2 11 DPM counting 2 11 DPM counting 2 11 1 Why DPM counting CPM counting is used when sample preparation is expected to yield samples with close to constant counting efficiency 1 e no variation in quench level This means that the results of the samples in an assay can be compared with each other and used in further data analysis If the counting efficiency varies with quench in the sample CPM counting can no longer give accurate resu
209. ny orders typed on the keyboard it is best to restart the program Turn the power off from the counter and the terminal Switch the power on again first the terminal and then the counter and try again If the counter does not restart then remove the battery fuse before switching the power on 2 13 2 Pressing the wrong keys If a wrong key is pressed the beep is heard and alternatives are shown again If in protocol editing a wrong value is given it can be easily erased by pressing the Backspace key and retyping the correct value If the Enter key has already been pressed then type L and the line number to return to the line to enter a new value or either the up arrow or P and Enter to go to the previous line 69 2 13 Errors 70 2 14 Half life 2 14 Half life 2 14 1 Radioactive half life The half life of a radioactive isotope is the time in which half of the radioactive nuclei have decayed 1 e the activity is halved The decrease in activity is exponential and can be calculated using the formula A 1 A 0 x2 T Where A 1 the activity at time t 1 A O the activity at time t 0 and t t 1 t O Thaif the half life of the isotope Each radioactive isotope has its specific half life If the half life of an isotope to be counted is short and if the counting time of a batch of samples is relatively long the last samples will have decayed more than the first ones so the results between earlier and later samples cannot
210. occurrence of PLATE will be sent to each medium although output selections would list different fields A cassette will be measured totally before the plate tables will be output 83 2 19 Microtitration plate format output 2 19 4 Examples Example 1 demonstrates the use of table format In the output selection PLATE SQP I CCPM1 CCPM1 the keyword PLATE Selects microtitration plate format A separate table is created for SQP I CCPM1 and ccpM1 values If the plate is of the 24 well type and all rows A B C D have been specified the resulting tables are full of 4x6 results Counting protocol nos 23 Fri 28 June 2000 15 11 Name PlateTESTI CPM normalization protocol no 40 Assay 7 Prot 33 7 Cass 7 Funce gt Cassette Nori 7 Shelf 3 4 6 SOP 1 l 2 2 4 5 6 A TDS 348 3 3929 35 2 94 37925 34851 B 2916 DANO l 29242 Dae Mg DOr PAG 54 e Sl DO 349 4 SO 348 6 Se eres 346 2 D 2904 1 DADA E O 2205 3 28923 2189 CCPM1 1 2 3 4 D 6 A 89950 89374 91991 Ol TLS ESTAS 91324 B 86066 75547 86783 76144 86622 74871 92202 90716 O22 14 91019 92012 OOTZS D 86131 TOLOI 86924 75498 86034 TSG CCPM1 1 2 5 4 3 6 A Liz IPE Tez 1 2 i ae dle B T2 S apa Teo lia T3 E 2 e eZ L2 R ae D Taz Tys beZ IETS tgz ken Total Count rates 20009321 CCPM Sf USLI 34 45 kBg End of assay 84 2 19 Microtitration plate format output Example 2 demonstrates how to include replicate statistics with the microtitration plate format output He
211. ocol The parameters are much the same as in the counting and normalization protocols with some exceptions Information whether normalization standardization has been done is given after the protocol name The following counting protocol parameters are not included in the standardization protocol 2 Counting mode 3 CPM normalization DPM standardization protocol 20 Count all positions in cassette 21 Cassette type Ale COURTING Control 42 Background correction The following parameters are new or changed when compared with the counting protocol 124 2 25 Protocols 2 Sample type Select normal or scintillation proximity assay for sample type ParaLux counting is set automatically if SPA is selected 3 Crosstalk correction Press Y to use crosstalk correction otherwise select N See chapter 2 7 Crosstalk correction for further information 14 Easy DPM Press Y to use Easy DPM quench curves Only the two DPM normalization samples are counted in standardization 20 Number of standards Give the number of DPM standards used for standardization The value must be in a range from 2 to 12 6 in dual label with 6 as default 21 Isotope 1 activity Give the isotope activity in DPM The range is from 1000 0 to 999 999 9 with 200 000 as the default value 23 Isotope 2 activity Give the Isotope 2 activity in DPM The range is from 1000 0 to 999 999 9 with 200 000 as the default value 24 Standard curve fit selectio
212. ocol 3 contains a generally used H normalization Normalization protocol 4 contains a generally used C normalization 214 2J 25 Protocols 2J 25 Protocols 2J 25 1 Four types of protocols A protocol is a set of parameters that controls the counting of samples There are four types CPM normalization protocol DPM standardization protocol MicroBeta Counting protocol MultiCalc Assay protocol A CPM normalization protocol contains parameters used when doing normalization including when using the JET features see chapter 2J 23 Normalization and a DPM standardization protocol contains parameters for standardization see chapter 2 12 DPM Standardization Standardization is not used with the JET feature A MicroBeta counting protocol is used when counting unknown samples An assay protocol is the corresponding protocol in MultiCalc see MultiCalc A counting protocol uses either a normalization protocol in CPM counting or a standardization protocol in DPM counting to get certain parameters e g isotopes and window settings Assay protocols are connected to counting protocols from which they get both basic parameters such as isotopes and counting mode and special features such as background and half life corrections The maximum number of protocols for each type is 100 0 to 99 Number 0 is the default protocol which is used if no protocol number is specified Counting protocol number 99 is a so called GLP Good Laborato
213. oefficient of variation The theoretical coefficient of variation CV of the mean M in percentages headings TCV 1 TCV2 is calculated using the equation TCV a 100 V gt 8A n M 31 3 3 9 5 Coefficient of variation The observed coefficient of variation CV of the mean M in percentage headings CV1 CV2 is calculated using the equation CV a 100V M A n 1 M 32 a 100V XA A n n 1 M 3 3 9 6 Theoretical standard deviation The theoretical standard deviation TSD headings TSD1 and TSD2 is calculated using the equation TSD a vV SA n 33 3 3 9 7 Observed standard deviation The observed standard deviation SD headings SD1 and SD2 is calculated using the equation SD VA LAn a 1 34 3 3 9 6 Chi square test The reduced chi square value Y headings CHI SQUARED CHISQ1 CHISQ2 is defined as Y O T 35 262 3 3 Calculation methods where the observed error O is from equation 30 and the theoretical error T is from equation 29 How much Y deviates from unity 1s a direct measure of the extent to which O deviates from T With a limited number of measurements we would not expect these two errors to be precisely the same Thus a more quantitative test 1s required to determine whether the observed difference between these two deviations 1s really significant For this purpose we can apply the chi square test From the y value we can calculate the probability headings P
214. om the counting of the two calibration standards mentioned earlier 63 2 12 DPM standardization It is preferable that each standard is counted with the same amount of pulses to get an accurate curve Set the counting precision on line 12 of the counting protocol Make sure the counting time is long enough that the precision requirement will terminate counting and not the counting time See chapter 2 25 Protocols section 2 25 7 number 12 for more details 2 12 1 4 Easy DPM If Easy DPM is selected then DPM standards are not needed The instrument uses stored quench curves for isotopes H sak I C a S and P Only two normalization samples are measured in standardization Normalization sample positions are H11 and H12 96 well plate 1 6 detectors G11 and G12 96 well plate 12 detectors D5 and D6 24 well plate P23 and P24 384 well plate 1 6 detectors M22 and M23 384 well plate 12 detectors 2 12 1 5 Evaluating unknown samples The actual sequence of events for counting unknown DPM samples is as follows 1 The SQP or AQP value and CPM are obtained with any detector 2 The SQP or AQP value is now corrected from the energy calibration curve to be what it would have been if the sample had been counted in detector 1 SGP for Energy calibration curve made with two points AIP 1 detector 1 Corrected SGP Cor AGP Measured SGP for APC SGPC Cor detector AIP 3 The corrected SQP I or AQP I is used to re
215. on plate Take sample solution with some activity scintillant which corresponds to the samples to be analyzed 1 e has the same isotope and solvent and pipette the same volume as in the samples into well G11 96 well plate or well N22 384 well 1 6 detectors or well M21 384 well 12 detectors For a 96 well plate with a 1 6 detector counter pipette background solution corresponding to the sample solution but without activity into wells Al F11 and G10 96 well where the latter two are the crosstalk samples G10 is only needed for strip plates and Al if background correction has been selected With a 12 detector counter use positions Al and F11 Strip plate is not available For a 384 well plate with a 1 6 detector counter use positions Al M22 and N21 for strip plate With a 12 detector counter use positions Al and M22 Strip plate is not available The sample array for a 96 well plate with a 1 6 detector counter is as follows where S1 is the sample solution and B1 C1 and C2 are the background solutions 41 2 Crosstalk correction TATUJI W gt Close the sample plate with a sealing tape and place the plate on a 1450 103 or 1450 105 cassette for counting 2 7 2 2 Preparation of normalization samples on a 24 well microtitration plate The procedure is the same as with 96 well plates but the normalization sample positions are D6 for the sample solution and Al C6 and D5 for the background solution D5 is only needed for
216. on is detected The LCPS out of range monitor is included in the short and long outputs in luminescence counting It cannot be used with the luminescence scaler board 93 2 21 Monitors and flags 2 21 5 Termination flag Counting is normally terminated when the specified counting time is reached or when the user stops it but in some cases it can be interrupted earlier Precision selected on line 12 in the protocol editor is such a case see chapters 2 25 Protocols and 2 6 Counting control The termination flag is set to PREC if the precision value has been reached The counting stops if the precision has been reached in all detectors Another case that terminates counting is maximum counts or hot sample If the count value in any channel in any detector exceeds a certain value usually 50000 counts the counting stops and the flag for the particular sample is set to MAX This prevents MCA channels from overflowing The flag is set to INT if the user interrupts the counting by selecting Off Next position or Next protocol Statistics monitor STM No statistical variation detected No variation after one recount Po REC No variation after two recounts ERR Still variation after two recounts DPM out of range monitor DPMM_____ OK no out of range detected OUT OutofDPMrange o O o LCPS out of range monitor LM_ OK no out of range detected 5 A a Luminescence scaler is used saturation cannot be detec
217. only the first 16 characters will be shown in the protocol list and press Enter The name may not start with characters or 2 Counting mode Type for CPM counting or 2 for DPM counting 3 CPM normalization protocol DPM standardization protocol Type the number for the normalization in CPM counting or standardization protocol in DPM counting used Type H or to get a list of normalization standardization protocols Isotopes must be entered if no protocol is selected otherwise the counting protocol uses the same isotope s as in the norm std protocol Type if no protocol is needed This means that no normalization correction is done in CPM counting and no DPMs are calculated in DPM counting Lines 4 and 5 are displayed only if no normalization or standardization protocol number is selected Information about whether norm std has been done is given after line 3 4 Number of labels Type 1 for single label counting one isotope default or 2 for dual label counting two isotopes 218 2J 25 Protocols 5 Isotope 1 Select an isotope in dual labelled samples the isotope with the lower energy The alternatives are 1 H default 2 T 3 4C 4 35g 5 Cr 6 P T 2P Cerenkov 8 Other 9 Luminescence You will be asked for window limits if you select Other otherwise window settings for the selected isotope are shown ParaLux counting can be selected in single label non crosstalk and non luminescence
218. onnected to the instrument RS232C port 3 See the manuals belonging to the printer about how to set up the printer for use with a serial interface Go to the Ter m inal selection in System state and set the line Print through terminal to N No do not change other lines Note See 4 8 6 for the default RS232C protocol and how to change it 4 1 8 2 2 Printer connected to the terminal PC parallel printer port As this is the default connection there is no need to change system parameters Proceed as follows a Use a standard Centronics interface cable to connect the printer and the terminal PC b See the manuals supplied with the printer about how to set up the printer for use with a parallel interface 4 1 8 3 Printer type Instrument Manual See chapter 2J 30 System P rinter Default Epson FX with printing resolution of 12 characters per inch cpi 298 4 1 Installation instructions The printer type selection affects only graphical output e g spectrum plots and the automatic printing resolution setting cpi The printing resolution affects the possible line lengths of outputs 10 cpi 80 12 cpi 96 and 17 cpi 136 character per line on a standard 8 inch paper The printer types supported are 4 1 8 3 1 IBM graphics printer Select this only if the printer does not support Epson FX emulation but supports IBM graphics printer emulation instead This mode does not support 12 cpi mode 4 1 8 3 2 Epson FX As this is
219. operation of MicroBeta TriLux P a sswords Print protocol passwords Customi z ing Set environment strings to customize the program Detector temp Set thermostat optional Test level contains also Serial n o Set instrument serial number Temp cali b Edit temperature calibration values Detector shelf u sage Set detector usage during counting set program support for the ParaLux board and set program support for the plate ID reader if installed and specify 1 16 32 shelf model 24 2 2 Cassettes 2 2 Cassettes 2 2 1 Construction A MicroBeta cassette 1s designed to hold a microplate and to allow it to be moved from the rack to the conveyor and back again There are three types of cassette that can be used in MicroBeta a 96 well a 24 well and a 384 well cassette If the instrument has six or less detectors it can take either a 96 or 24 well cassette or a 96 or 384 well cassette depending on the mask used If it has twelve detectors then only 96 well and 384 well cassettes can be used Canas fae e aC AH i 450 140 a a Cassette 1450 101 This has 96 sample holes with a diameter of 7 8mm The cassette is used when counting 1450 401 or equivalent 96 well sample plates Cassette 1450 102 This has 24 sample holes with a diameter of 13 2 mm see the figure This cassette is used when counting 1450 402 or equivalent 24 well sample plates Cassette 1450 103 This has 96 sample holes The cassette is used when counti
220. or other filters Melting temperature 60 120 C 1450 442 MeltiLex B HS 50 box Meltable solid scintillator for filters 1450 521 1450 424 and other thick filters Melting temperature 60 120 C 1450 444 Polystyrene Safe 5L box Scintillation cocktail to be used with polystyrene sample plates e g 1450 405 1450 407 and 1450 410 274 3 4 Specifications 3 4 15 5 ID Products 1450 451 ID Support Plate Support plate for ID labels Placed onto a counting cassette 1450 452 ID Labels 10 sheets box Labels for identifying Assay number necessary for MultiCalc assays Protocol and Cassette number and Function code Fixed on a Support Plate 1450 451 3 4 15 6 Tapes 1450 461 Sealing Tape 100 box Sealing tape for sample plates permanent 1450 462 Removable Sealing Tape 100 box Sealing tape for sample plates removable 1450 463 Heatsealing Foil 100 box Sealing foil for sample plates type 1450 401 and 1450 408 Heatsealed with a 1495 021 Microsealer 1450 465 Printed Sealing Tape 100 box Sealing tape for sample plates printed 1450 466 Coloured Sealing Tape 100 box Sealing tape for luminescent sample plates 3 4 15 7 Other Consumables 1450 481 Support Frame 25 box Support frame for sample plates type 1450 401 1450 402 and 1450 408 Removed before counting 1450 482 Support Frame for 1450 407 25 box Support frame for sample plates type 1450 407 and 1450 419 Removed before counting 1450 407 1450 4
221. or precision value Note that this is expressed as 1 sigma percent while the precision in the counter is expressed as 2 sigma percent Select the isotope s on line 03 MEASURING PARAMETERS using one of the softkeys Fl to F8 The isotope names can also be typed in The Counting protocol to be used corresponds to the selected isotope as shown in the table following 98 2 22 MultiCalc operation Counting protocol The Counting protocols 91 to 98 are factory made They may not be deleted and the isotopes may not be changed They are connected to factory made CPM normalization protocols with the same numbering 91 98 Note No CPM normalization or DPM standardization is made at the factory so you must prepare suitable normalizations or standardizations for the factory made counting protocols The Counting protocol number can also be typed instead of giving the isotope Please check that this protocol exists and that it has the right isotope s and counting mode If the protocol does not exist the default protocol number 0 will be used Examples of selections on line 03 H Counting protocol 91 13 useda HS CLA Dual label Counting protocol 96 H C is used 18 Counting protocol 18 is used can be single or dual label If you are using a newer version of MultiCalc you will first be asked for the counter instead of the isotope Press softkey PLATEV3 A list of counting protocols will be displayed and the isotopes available w
222. ored into the directory which was active when GenTerm was started 4 full disk drive and directory path Use this if options 1 3 do not match your requirements E g if you want to store results a on a hard disk drive other than C e g D b on a local area network LAN connected to the terminal PC c in a specific directory or d on the MicroBeta drive If 4 1s selected then the program asks for the full drive and directory path Here understanding of MS DOS drive and directory concepts is needed The directory path given here will be used as such without any checking and modifications in front of the file name Note that the directories must exist before any data can be sent to them Use the command MKDIR in DOS to make new directories Examples referring to examples a d above a Path name D gt Text D will be inserted in front of result file name E g D 2 003 the file will be stored in the active directory on drive D b Path name F gt Here it is supposed that drive F belongs to the LAN server Text F will be inserted in front of the result file name E g F 17 002 the file will be stored in the active directory of the LAN server drive F c Path name C MB RESULTS gt Text C MB RESULTS will be inserted in front of result file name Note that if you specify an exact directory path for the result files then the path name must end with backslash For example the path c MB RESULTS
223. ormation whether normalization standardization has been done is given after the protocol name The following counting protocol parameters are not included in the standardization protocol 2 Counting mode 3 CPM normalization DPM standardization protocol 20 Count all positions in cassette 21 Cassette type 41 Counting control 42 Background correction The following parameters are new or changed when compared with the counting protocol 2 Sample type Select normal or scintillation proximity assay for sample type ParaLux counting is set automatically if SPA is selected 3 Crosstalk correction Press Y to use crosstalk correction otherwise select N See chapter 2 7 Crosstalk correction for further information 14 Easy DPM Press Y to use Easy DPM quench curves Only the two DPM normalization samples are counted in standardization 20 Number of standards Give the number of DPM standards used for standardization The value must be in a range from 2 to 12 6 in dual label with 6 as default 21 Isotope 1 activity Give the isotope activity in DPM The range is from 1000 0 to 999 999 9 with 200 000 as the default value 23 Isotope 2 activity Give the Isotope 2 activity in DPM The range 1s from 1000 0 to 999 999 9 with 200 000 as the default value 229 2J 25 Protocols 24 Standard curve fit selection Press Y to use standard curve fitting otherwise N If No is selected smoothing spline with automatic smoo
224. orrection Y N Use normalization background Background sample Y N Y N Bgnd sample position G Bgnd counting time s Half life correction Y N Chemiluminescence correction Y N Use password Y N Special plate 1 Wallac 2 BP filter 3 Other Strip plate Y N 67 i bmp aa a ON ee Zak ZP gt gt gt gt gt gt gt gt gt gt gt gt Activity of standards is given on line 21 gt Y Curve fitting method given here gt gt gt f Y Y gt gt Y gt gt gt gt Background sample selected gt 2 12 DPM standardization 2 12 4 Preparing dual label standardization samples Two plates are used and the higher energy isotope samples are counted first A maximum of 6 DPM standards can be used on each plate For 24 well plates the sample positions are the same as in single label standardization For a 96 well plate the standards are placed in the following positions on each plate B1 B3 B11 the first normalization standard in position G9 96 well 1 6 det G10 96 well 12 det and the last normalization standard in position G12 96 well 1 12 det 2 12 5 DPM standardization procedure The plate s with the standardization samples are placed in cassette s which are identified with the corresponding standardization protocol number in the protocol number area and with a STD label in the function code a
225. ould be used if you want to edit normalizations standardizations and counting protocols from within MultiCalc With a terminal 2 37 you can get results to your screen print Results are sent The printer Output Local Results go to PC them out or store them on a disk to MicroBeta connected to to area connected to port in MicroBeta drive B during MicroBeta port external net 1 in terminal A terminal emulator 2 3 is a terminal 3 is used for compu work emulation amp program which makes a PC act operation For terminal and tere ga conn MultiCalc See as a terminal You can have diskette terminal main ection 2 10 for diskette results stored on your PC You information see emulation frame LAN info MultiCalc can also use your PC for other 2 10 printing Con using printout goes to tasks during counting nection the PC the printer _ to port connected to the A E For System setting info e g 2 or PC PC Entering commands into printer or terminal type 2 30 MicroBeta is described in 2 1 For Help type H h or 2 15 MultiCalc has its own manual 10 Operating MicroBeta JET See chapters in italics for more information on any subject barcodes are fixed Cassette 1450 101 to an ID support plate which is clipped onto a cassette to show MicroBeta the counting protocol to use 2 4 A counting protocol lis a set of unction code parameters e g time isotope single dual label
226. ount C opy P rotocols S ystem PM norm protocol gt Help Q uit E dit nn prot no D elete Protocols gt S how Help P rint Q uit L ist Injector C ounting protocol De f ault H elp CPM n ormalization prot C opy Q uit DPM s tandardization prot N ormalization data Dnit M ultiCalc assay protocol eee D ispense DPM stand protocol gt P rime H elp T ip System gt Q uit W ash H elp E dit Suck b ack Q uit nn prot no Co m mand MS D O S D elete S tatus E rror beep S how of S tatus display P rint N ext pos V ersion L ist G oto pos C lock De f ault C lear conveyor Se t up mode C opy Disp e nser IN OUT R S 232C R eplot P rinter D ata drive ultiCalc assay protocol gt Ter m inal H elp P a sswords Q uit Customi z ing S how Detector temp P rint L ist MicroBeta A gt MB Back to Ready gt 192 2J 1 Beginning operation of MicroBeta JET MicroBeta JET commands normal level with a brief description of each Operations in the Ready State H elp Display Help text Dnfo Get information on using the instrument Cjount Start counting or operate conveyor P rotocols Edit counting parameters S ystem Edit system parameters Count state operations are available in the Ready state too Operations in the Info State Help Display Help text Q uit Back to the Ready state U sage View a Short user s manual C onsumables View a list of consumables a
227. pair works in coincidence The tubes of the pair are situated on opposite sides of the counting block one above and one below the block The detectors form a 4 x 3 array see the figure below 7 A2 nO A A7 A8 Ag A11 A12 Bl E2 B63 Bo BS B 67 68 BY BIA E11 BI C1 C2 C3 GS CS C CF CGH CY 1168 C11 C12 b7 n2 D3 0a DS b 1 pe D9 Em D11 D12 model cassette in first position El E ES Eb ES Eb E EB ED EIA EW El Fl F2 F3 FH FS F Fe FR FO F10 F11 F12 Gi 62 63 6x 65 66 67 68 69 6 611 612 H1 H2 H3 H4 HS H H7 WE H9 H16 H11 H12 Twelve detector Twelve samples are counted simultaneously which reduces counting time to about one twelfth of that for a single detector instrument There are nine readings per plate When a cassette is being counted the transportation system moves it between the detector blocks The first detector then counts position A1 the second A4 etc At this phase only the results from position A1 are output because the results must be in the order A1 A2 A3 etc The cassette moves one position forward and positions A2 A5 etc are counted The results from A2 are output After counting the whole row the cassette moves to the beginning of the next row e g position B1 in detector 1 The rest of the results from row A are output A limited range of positions 10 to 12 in row G can be counted by all detectors This affects the positioning of CPM normalization and DPM standardization samples
228. pe 1450 104 1450 522 DEAE Filtermat 100 box 96 position filter mat with printed pattern made of glassfibre containing positively charged groups For harvested or pipetted negatively charged samples Scintillant to be used Betaplate Scint 1205 440 or MeltiLex A 1450 441 Placed in sample bag type 1450 432 Counted in cassette type 1450 104 1450 3523 P30 Filtermat 100 box 96 position filter mat with printed pattern made of glassfibre containing negatively charged groups For harvested or pipetted positively charged samples Scintillant to be used Betaplate Scint 1205 440 or MeltiLex A 1450 441 Placed ina sample bag 1450 432 Counted in a cassette type 1450 104 3 4 15 3 Plastic Bags 1450 431 Waste Bag 100 box Waste bag for used sample plates Chemically resistant to safe cocktails Closed with a heatsealer 1295 012 1450 432 Sample Bag 100 box Sample bag for filters 1450 421 and 1450 423 Chemically resistant to safe cocktails Closed with a heatsealer 1295 012 3 4 15 4 Scintillation Products 1450 419 ScintiStrip 120 box 12 breakable strips of 8 wells on support frame type 1450 482 made of scintillation plastics max volume 300 uL well For immobilised samples or aqueous samples no scintillation cocktail needed Sealed with tape type 1450 461 or 1450 462 Counted on the support frame in cassette type 1450 105 1450 441 MeltiLex A 100 box Meltable solid scintillator for filters 1450 421 and 1450 423
229. pearing Continue until the cursor blinks just after CTIMR _ shows the place of the cursor POS CEMR 17 2 1 Beginning operation of MicroBeta TriLux If you go too far then stop pressing Control B and press Control N to make the lost characters visible again When the cursor is placed just after CTIMR as in the example press Backspace or the DEL key once and R will disappear Type the letter E gt POS CTIM_ Backspace or DEL pressed gt POS CTIME_ Letter E typed Keep pressing Control N until there is a beep and you get gt POS CTIME COUNTS1 CCPM1 CCPM1 You can now continue typing new headings after CCPM1 E g gt POS CTIME COUNTS1 CCPM1 CCPM1 MEAN1 CV1 Now press Enter to complete editing If you later want to remove the heading COUNTSI go to line 31 to get the programmable output selection Keep pressing Control N until you are just past COUNTSI gt POS CTIME COUNTS1_ Press Backspace or DEL 8 times so that COUNTSI and the preceding space disappears gt POS CTIME_ Keep pressing Control N until there is a beep and press Enter gt POS CTIME CCPM1 CCPM1 MEAN1 CV1 _ You can also add a new heading afterwards Go to programmable output selection again Keep pressing Control N until the cursor is just after CTIME o POS WIM eE Now type space and the text SQP I gt POS SI LME SOPII Keep pressing Control N until the program beeps and then press Enter gt POS CTIME SOP I CCPML
230. placed with ppm in DPM counting In luminescence counting itis POS LCPS and LCPSS 3 Long The long output contains PROT POS CTIME SQP I Or AQP 1 CCPM1 CCPM1 plus CCPM2 CCPM2 in dual label counting In DPM counting DPM1 and DPM1 are added In luminescence counting the output is PROT STIME POS CTIME CPS1 LCPS LCPS 4 Programmable The output items and arithmetic expressions are defined separately on the next line Sample quality monitors can also be selected in the programmable output Press or H to get a list of output items monitors and rules for using own formulae See also chapters 2 1 Beginning operation 2 26 Results 2 29 Statistics and 2 21 Monitors and flags If SPECTRA 1s selected three more lines are shown The first asks for the energy scale to be selected It can be logarithmic with the units in channels or linear with the units in keV or both of these Next you are asked to specify the range of the plot This will be channels or keV depending on your previous selection Finally you are asked for the spectrum size small medium or large A smaller size will give faster output The two figures following illustrate the type of plots you may get 118 2 25 Protocols POS CTIME COPM1 CRRMIX AG 8 7 214514 1 O 4 Counts Scale 1 4 Ayla a Al VA 100 200 BOG 400 TOG 500 FOO Channel Spectrum with logarithmic scale O E 40 E T 100 120 1913 Lao 180 Spectrum with linear s
231. plates Prepare a series of quenched OptiPhase SuperMix samples E g dispense into 3 ml samples of OptiPhase SuperMix the following amounts of carbon tetrachloride 0 10 30 50 75 and 100 uL Mix these samples Next dispense the activity to wells Al to A6 G11 and G12 then dispense 200 uL of the quenched cocktail to each well Seal the plate and count it as described in chapter 2 11 The activity used should be e g 20 uL of tritiated water or some other suitable source giving about 200 000 DPM per well so that the counting time required to get an acceptable precision e g 0 5 can be about 1 min If you do not mind using a longer counting time you can use lower activities 88 2 20 Micro volume LSC Note The total time for a standardization with 200 000 DPM activity would be less than 20 mins whereas for 20 000 DPM it would be 3 hrs to get the same precision 2 20 6 Chromium release Specific cytotoxicity can be measured with microvolume samples by counting the supernatant from the chromium release Add 25 uL of supernatant to 175 uL of OptiPhase SuperMix cocktail The crosstalk between wells is of the order of 0 06 when using the printed microtitration plates Use a counting window setting of 5 170 2 20 7 Microvolume LSC and chromatography A chromatography eluent gradient with varying ionic strength can be collected in 96 well plates for evaluation in MicroBeta The different aliquots will have different levels of quenching a
232. r The characters you type are not displayed on the screen If you enter a wrong password the program displays the message Incorrect password Otherwise the program shows the full System menu To set a system password exit from the MicroBeta program to MS DOS as described earlier After the text MicroBeta A gt appears type PASSWORD xxxx and press Enter xxxx in the above means the new password If you forget the password then set a new one 157 2 30 System 158 2 31 Terminal emulators 2 01 Terminal emulators 2 31 1 Introduction MicroBeta can be used with a variety of different terminals and terminal emulator programs Most of the chapters in part 2 of this manual describe terminal operations The only exception is when the MultiCalc counter program is used This chapter describes operation with two general purpose terminal emulator programs from Wallac GenTerm and UltroTerm and VT52 and VT100 terminal emulators see below for an explanation of the terminology For information about MultiCalc see chapter 2 22 MultiCalc You can select whichever of these terminal emulations is most suitable for your needs 2 31 2 Terminology 2 1 2 1 Terminal This consists of a keyboard display and electronics and is not usable as a general purpose computer 2 31 2 2 Terminal PC This 1s any general purpose personal computer that can run a terminal emulator program 2 31 2 3 Terminal emulator program This 1s a program which allow
233. r ParaLux counting and efficiency SQP I or AQP is a measure of the isotope spectrum and varies as the quench changes Once this relationship between SQP I or AQP and efficiency is known it is possible to calculate the counting efficiency for any measured SQP I or AQP I value in the range covered by the measurement of the standard samples Knowing the efficiency the DPM can be calculated 2 12 1 2 Detector energy and efficiency calibration However closely matched detectors are there will be some difference in their energy calibration affecting SQPC or AQP I measurement and counting efficiency A correction must be made for both these differences 2 12 1 3 Standardization Standardization Detector energy calibration Sample quench correction detector efficiency calibration In MicroBeta the standardization is done by first measuring a maximum of 12 DPM standards with defined activities in detector 1 and then counting two DPM standards in every detector The two calibration standards should be representative of the range covered by the DPM standards 1 e they could be replicates of the first and last DPM standards The quench standard measurement produces a curve of efficiency versus SQP I or AQP with as many points as standardization samples Energy calibration and efficiency calibration are both linear functions so two points are sufficient to define these two calibration curves The two points are obtained fr
234. r cassettes do not have ID labels simply type the number of the protocol to be used and press Enter The number you enter must be within the allowed range 0 99 Pressing brings a list of protocols to select from If counting was started with an A command and the first cassette has no counting protocol code then the first assay will be counted using the default counting protocol protocol number 0 Any other cassette that does not have a counting protocol number will be counted with the protocol defined by the most recent cassette with a counting protocol number This means that within one assay you only need to label the first cassette A new label is only needed to start a new assay 2 5 1 2 Automatic normalization or standardization terminal operation If your cassettes have normalization or standardization protocol and function ID labels press A The counting protocol to be used will be determined by the ID labels If your cassettes do not have ID labels type the protocol number preceded by N in normalization e g N15 and D in DPM standardization e g D15 Typing N or D brings a list of normalization or standardization protocols to select from Normalization or standardization can be done before sample counting See the chapters 2 12 and 2 21 33 2 5 Counting 2 5 1 3 Automatic counting MultiCalc Your cassettes must have Assay ID labels on them Starting from the main MultiCalc menu press Fl COUNTER and then
235. r output The default programmable output PRINTER means the same data as specified for printer output 34 External output This output is selected when sending data to an external device such as a mainframe computer connected to the MicroBeta via port 2 The data is in ASCII format text files The output alternatives are the same as in printer and display output except that alternative 1 No output is the default value The default programmable output PRINTER means that the same data is specified as for printer output 36 File output The counting results can be stored on a floppy disk or hard disk on the PC or MicroBeta or on the server of a local area network LAN section 2 8 Datafiles The default setting is 1 no file output The alternatives are the same as in printer output The default programmable output PRINTER means that the same data is specified as for printer output The result files are named so that the first part of the name the part before the period 1s the protocol number and the second part after the period is a running number indicating the assay s execution order E g the first file for protocol 5 1s named 5 001 the second 5 002 etc Result file names for normalizations begin with N e g N1 001 and standardizations with an S e g S1 001 The disk drive A B for floppy disks C and D for hard disks F etc for LAN drives and the path e g MB DATA for the result files are chosen af
236. r yes and then the DPM value A background sample is counted only 1f selected on line 42 If you use strip plates then select Y on line 47 The crosstalk sample in G10 D5 or N21 is counted only if strip plate is selected 2 7 2 4 Normalization counting Start the normalization run in the Ready or Count state using automatic counting and ID codes or by selecting Nx where x is the number of the protocol The normalization sample is then counted in every detector and the crosstalk sample in detector l 2 7 2 5 Counting the actual samples using crosstalk correction Edit a counting protocol Select in the Ready state P P rotocol then C C ounting protocols Then select E E dit and a protocol number 43 2 Crosstalk correction On parameter line 2 select 1 CPM and on line 3 select the number of the crosstalk normalization protocol defined above Edit the other parameters normally Start counting using automatic counting and a protocol ID label or start with the protocol number and press Enter When counting the samples the stored crosstalk factors are used for correcting the CPM values The crosstalk corrected CPM values are marked as CCPM1 in the printout 2 7 3 Crosstalk DPM standardization 2 3 1 Preparation of DPM standardization samples on a 96 microtitration plate 1 6 detector counter Make first the standard samples in vials and then pipette them onto a plate a 6 vials for isotope standard samples and 6 v
237. re the keyword stat can be used see also the example in section 2 29 3 3 Statistics File output The printout fields defined after strat will be output only when a replicate group is ready With the plate format output this means that the printout defined after STAT will not be represented in table format but as normal output In addition results will be output at once so the statistics will appear in output before the plate format raw data In Example 2 the output selection PLATE CCPM1 CCPM2 STAT REPL MEAN1 CV1 MEAN2 CV2 s1 can be interpreted as PLATE Use microtitration plate format Collect ccpm1 and ccpm2 values into tables and output those tables when the cassette has been measured STAT Do not collect the following printouts into tables but output them when the replicate mean value is available REPL Output replicate range e g AO1 A03 MEAN1 CV2 Output these statistical values S1 Output deviation values cv1 CV2 at the 1 sigma level Counting protocol noo Fri Ze June 2000 15 22 Name PlateTEST2 CPM normalization protocol no 40 Assays Prot 23 7 Casi Funes Cassette Norl y Shelty 3 1A 6 REPL MEAN CV1 MEAN2 CV2 A01 A03 ZO SOO a J LD 975249 Ose A04 A06 3160973 Ss TODO 0 8 BO I B0 3 61304 6 206 ES PES BO4 B06 Ge693 lt 1 Loi S509 26 rie COM SCOS LISO 14 4 97421 2 Olas C04 C06 32664 4 14 0 96547 0 O DOT D03 6270850 LS SL Li al 6 8 DO4 D06 69314 6 16 4 SITIO OO Seo CCP ML 1 2
238. rea In dual label standardization the first higher energy isotope cassette should have the STD label or STD2 for the first plate and STD1 for the second Insert the cassette s into the rack and close the door Press either A Automatic counting or Dxx where xx 1s the standardization protocol number in the Ready or Count state to start counting If Dxx 1s selected the program asks Do you want to continue counting after standardization Y N gt If Y Yes is selected the counting continues after counting the standardization plate if N No is selected the counting stops after the standardization If standardization has already been done for the protocol then the program asks first This protocol contains standardization data Continue Y N gt This is to prevent accidental overwriting of standardization data If automatic counting is selected the standardization cassette can be placed anywhere in the rack The STD label is needed in this case The counting results for the standardization are output as specified on lines 30 37 in the standardization protocol 2 12 5 Standardization results When a standardization is done the results are stored with the standardization protocol The standardization data can be used by one or several counting protocols and is selected when editing the protocol see chapter 2 25 Protocols 68 2 13 Errors 2 13 Errors 2 13 1 Program failure If the program is jammed and will not take a
239. rmation about protocols 2 4 3 Stop cassette If you fix a STOP code to the last cassette it means that this cassette will be counted also However it can be tedious to remove this STOP code and stick it to another cassette when more cassettes are loaded To avoid this a stop cassette can be used Stick a STOP code on both the protocol and function code field of an empty cassette and put that cassette after the last cassette to be counted 2 4 4 Cassette operation If MicroBeta is loaded with many sample cassettes that are to be counted with different protocols the cassettes are coded with the respective protocol number If there is no protocol number on the cassette the cassette is counted with the same protocol as the previous one 1 e it 30 2 4 Coding cassettes and filtermats is considered to be part of the same assay To stop the counting automatically the STOP code should be used 2 4 5 Filtermat coding 123456789 0 n 126 DAY 345 672 89 W112 3 4 5 18328128 11666686666 000 On the lower edge of the filtermat there are two groups of twelve small circles see the figure They are for marking the date This is done by cutting or clipping off appropriate circles The first twelve circles are to specify the day and the second twelve the month In the figure above the leftmost 9 circles stand for the numbers from to 9 and the next three circles for 10 20 and 30 respectively E g the 25th day is marked by cutting off
240. roBeta works its defaults hardware connections etc To enter the System state press S in the Ready state You are then given a number of options as example shows These are described in more detail here Example 1 H elp I nfo C ount P rotocols S ystem Ready gt s H elp O TE MS D 0 S E rrorbeep S tatus display V ersion C lock Ry GHZ 326 P rinter D ata drive Ter m inal P a sswords Customi z ing 1 jector module System gt 2J 30 2 H elp Press H to get help about the various selections 2J 30 3 E rror beep User or instrument error normally causes a beep This can be switched off by selecting N Example 2 System gt E Error be p Y N Y gt N 2J 30 4 S tatus display Normally when counting is occurring its status is shown on the Terminal PC screen see chapter 2 5 Counting for more details The counting status display can be switched off or only currently counted positions can be shown Example 3 system gt s Status display 1 No 2 Current positions 3 Whole plate gt 233 2J 30 System 2J 30 5 V ersion Show the program and MS DOS version numbers Example 4 system gt V MicroBeta program V5 0 Copyright C Wallac Oy 1998 All rights reserved MS DOS Vas COpyragne E MECEOSOTE Corporation 1987 All rights reserved 2J 30 6 C lock This allows the date and time to be set MicroBeta has a calendar clock with battery back up so you do not have to set it each time the pow
241. round sample is measured then the count rate a sigma error becomes 256 3 3 Calculation methods o Sr r av 81 SD r 6 3 3 3 2 Detector efficiency correction In single label counting detector efficiency correction for count rate is calculated using the equation R r e 7 where r is from equation 3 and e e 11 from equation 4 for the same detector as in normalization Efficiency correction affects also the corrected count rate a sigma error o 8R R av Sr r Se e 8 where 0e e de 11 e 11 from equation 5 and or r is from equation 6 In dual label counting the efficiency correction has the form R m e nn r m e mn r n d 9 where d e mm e nn e nm e mn 10 and e mn stands for isotope n efficiency in window m Corrected count rate sigma percentage error in the dual label case is calculated using the equation o dRGn RGn o 1007 rm 6 Se nm y e nmy j Sr m r ny l Se mm e mm 8r n p m Q d 11 where r m is from equation 3 or m is from equation 6 d is from equation 10 p m e nn r m e nm r n 12 25 3 3 Calculation methods and Q e mn Se nm e nm r Se mn e nny dl Se mm y e mm Senn 13 3 3 3 3 Half life correction Half life corrected count rate I is calculated using the equation L t t In 2 exp In 2 T t t exp In 2 T t R 14 where t is counting time T
242. ry Practice protocol Count the normalization sample plate supplied with the instrument using this protocol Do this from time to time to make sure that the instrument produces consistent results The background H and C samples are counted with all detectors so background level CPM values and quench parameter values can easily be compared See chapter 3 2 Routine maintenance for more details In this chapter only MicroBeta protocols are described For further information about assay protocols see chapter 2 22 MultiCalc and the MultiCalc User manual Protocols 2J 21 2 Listing protocols To list or edit protocols press P and then C for counting protocols N for normalization protocols or S for standardization protocols in the Ready state To select the MultiCalc assay functions as described in chapter 2 22 press M 215 2J 25 Protocols To display the protocol list in the Protocols menu press S Show You will see a two column list of protocols containing protocol number name if given and isotopes The list of the counting protocols also contains counting mode CPM or DPM and the number for the normalization or standardization protocol used The lists of the normalization and standardization protocols have a column containing the date when the normalization or standardization has been done An equals character indicates that crosstalk correction is used If there are more protocols than there is space for on the display press En
243. s 2J 18 2 Preparation of glow type normalization samples 96 and 384 well format Make a sample solution which corresponds to the samples to be analyzed 1 e has the same enzyme and substrate and pipette the same volume as in the samples into well membrane G11 96 well 1 6 det N22 384 well 1 6 det An empty background plate is used for background measurement 201 2J 18 Luminescence counting The 96 well sample array is as follows where S1 is the sample solution l 2 3 4 5 6 7 8 9 10 11 12 TAHIA W D gt Plate Close the sample plate with 1450 461 sealing tape or with 1450 466 coloured sealing tape and place on a 1450 105 cassette for counting Filter Enclose the filter in a 1450 432 plastic sample bag and place it on a 1450 104 cassette use 1450 466 coloured sealing tape between the sample bag and detector 2J 18 3 Preparation of glow type normalization samples 24 well format The procedure is the same as with a 96 position sample support but the normalization sample position is D5 for S1 J awe 2J 18 4 Preparation of flash type normalization samples 96 well format Standards are arranged in blocks on the normalization plate If background sample measurement is used then the background sample must be in the first position in each block on the plate see the plate maps following 202 2J 18 Luminescence counting The plate maps for different detector configurations with 96 well plates and th
244. s E9 E10 E11 E12 first position F1 F F3 FA F5 F F7 F8 F9 F10 F11 F12 G1 G G3 G4 65 G G7 G8 G9 G10 G11 G12 H1 H H3 HA HS H H7 H8 H H10 H11 H12 Only the first detector can count every position in a plate The following table shows which positions can be counted by each detector 53 2 9 Detectors Det No 96 well 24 well 384 well 1 every every every 2 A5 A12 A3 A6 A9 A24 Ho H12 DS D6 P9 PZ24 3 A9 A12 A5 A6 A17 A24 HS H 12 Do D6 Piv P24 4 ETSET2 GL C6 11 124 OEA Di D6 o esa HLSHT2 Pp1 P24 5 ES EL2Z Eno 19 124 ss D3 D6 aid HOSHII P9 P24 6 E9 E12 EN 117 124 lia regs Do D6 alaba AS9 H12 Pp17 P24 The smallest range of positions is that counted by detector 6 This is therefore the only range that can be counted by all detectors This affects the positioning of CPM normalization and DPM standardization samples because these routines involve every detector counting the same sample See chapters 2 12 and 2 23 for details 54 2 9 2 Three two and one detector models 2 9 Detectors The MicroBeta series also includes models with 3 2 and detectors The detectors are then arranged as shown in the following figures 49 a2 B1 01 D1 E1 F1 G1 H1 B2 C D2 E F2 G2 H2 At a2 B1 B2 c1 C2 D1 D2 E2 F1 F2 G1 G2 H1 H2 D a2 B1 01 D1 E1 F1 G1 H1 B2 C2 Db E F2 G2 H2 A3 B3 C3 D3 E3 F3 G3 H3 AY Ay Ba C4 D4 En F4 G4 Ha BS AS BS C5 D5 ES
245. s a PC to be used as a terminal and often implements extra features such as data logging multiple terminal emulations etc 2 31 2 4 Terminal emulation This 1s a control language that can be used to command the terminal to perform simple operations such as clear display move cursor use underline etc The control language consists usually of plain ASCII text control codes such as backspace and escape sequences character sequences started by the Esc character Terminal manufacturers have introduced several different terminal emulation languages e g VT52 VT100 and Wallac WT 2 31 2 5 MS DOS MS DOS means Microsoft Disk Operating System a product of Microsoft Corporation MS DOS enables the user to operate on disks files directories execute programs etc The MicroBeta program itself and usually the terminal emulator program run under MS DOS When you exit from the MicroBeta program to MS DOS which is running in the instrument microcomputer you see the prompt text MicroBeta A gt In contrast when you exit from the 159 2 31 Terminal emulators terminal emulator program to the MS DOS of the terminal PC the prompt usually shows either the current disk drive e g A gt or the current directory e g C GENTERM gt For details about using MS DOS see the MS DOS User s Manual of your terminal PC 2 31 2 6 Temporary exit from a terminal emulator program Temporary exit from a program means that the current program remains in the
246. s and the counts observed in the bottom PM tube In other words it establishes a relation between quench and efficiency 2 20 9 2 Counting modes ParaLux counting is set in the protocol at the same time as the isotope is selected see chapter 2 25 Protocols ParaLux counting has two modes Low background mode default and High efficiency mode The difference between them 1s as follows In the Low background mode the count rate is calculated from just the coincidence events In the High efficiency mode the count rate is calculated from the coincidence events plus the events not in coincidence that are detected in the lower tube However these latter events are only registered for pulses that come in channels exceeding the discrimination channel set for the MCA You can set this discriminator channel level the default is 150 Both modes use AQPQ I as the quench parameter Both modes give improved counting performance because of the use of the Asymmetric quench parameter The High efficiency mode gives a higher efficiency but also a higher background than the other mode 91 2 20 Micro volume LSC 92 2 21 Monitors and flags 2 21 Monitors and flags 2 21 1 Functions of monitors and flags Monitors are used to check the quality of samples and flags to notify of e g poor sample quality or that counting has been terminated They are selected in the programmable output in protocol editor see chapter 2 25 Protocols The flag is include
247. s the luminescence counting background because the cooling decreases the photomultiplier tube thermal background E g changing the detector temperature from 27 C to 20 C decreases the thermal background by about 60 The thermostat option can be used to cool the upper detectors up to 9 C below room temperature The detector temperature can be selected in the System state by using the menu selection Detector temp System settings with the Thermostat option H elp I nfo C ount P rotocols S ystem Ready gt s H elp OVULE MS D O S E rror beep Status display V ersion Cy Lock RS Z 32 P rinter D ata drive Ter m inal P a sswords a ustomi z ing Detector temp system gt Example of how to use the Thermostat option SyYstem gt Current detector temperature 25 2 C Detector temperature C 24 0 gt 20 Control detector temperature Y N N gt Y Detector cooling can cause water to condense inside the instrument if the 0 relative humidity is above 73 When the detectors are cooled water can condense inside the instrument depending on room temperature detector temperature and relative humidity Please check that the relative humidity is below the number given in the warning text Detector temperature can be selected to be part of programmable output by using the heading TD 175 2 32 Thermostat option 176 2 33 Total count rate 2 33 Total count rate A total count rate r
248. say protocol is the corresponding protocol in MultiCalc see MultiCalc A counting protocol uses either a normalization protocol in CPM counting or a standardization protocol in DPM counting to get certain parameters e g isotopes and window settings Assay protocols are connected to counting protocols from which they get both basic parameters such as isotopes and counting mode and special features such as background and half life corrections The maximum number of protocols for each type is 100 0 to 99 Number 0 is the default protocol which is used if no protocol number is specified Counting protocol number 99 is a so called GLP Good Laboratory Practice protocol Count the normalization sample plate supplied with the instrument using this protocol Do this from time to time to make sure that the instrument produces consistent results The background H and C samples are counted with all detectors so background level CPM values and quench parameter values can easily be compared See chapter 3 2 Routine maintenance for more details In this chapter only MicroBeta protocols are described For further information about assay protocols see chapter 2 22 MultiCalc and the MultiCalc User manual Protocols 2 21 2 Listing protocols To list or edit protocols press P and then C for counting protocols N for normalization protocols or S for standardization protocols in the Ready state To select the MultiCalc assay functions as described
249. sc key to get to the UltroTerm main menu 2 Use the down arrow key to go to Set RS232 comm s protocol and press Enter 3 Enter communication port number and press Enter 4 Select an item from the Set Up menu by pressing the up or down arrow keys 5 Change the value of the selected item by typing the new value or by selecting from the list using the Home and End keys 6 Confirm the change by pressing Enter 7 Store the setup on disk as default settings by answering Y es to the last question 166 2 31 Terminal emulators 8 Press Esc twice to return to the terminal emulator mode Section 2 30 System R S 232C in this manual describes how to change communication parameters in the counter This section also contains the default parameter settings 2 31 5 6 Special features UltroTerm uses an extended VT52 terminal emulation UltroTerm allows file transfer to a computer built in file editing and printing 2 31 5 7 Restrictions UltroTerm can serve only one instrument when in 1205 mode In 1205 mode MicroBeta controls printing so the F7 key does not work 2 31 5 8 Result files Select Auto filing start string and Auto filing stop string in Filing parameters A column selection marker e g can be used with programmable output see 2 26 3 See the UltroTerm User Manual for details 2 01 5 9 Printer connection The MicroBeta program can automatically direct printer output through UltroTerm to the printer connected
250. ser must check and correct the terminal and or RS settings in System parameters The program will usually work fine with the automatic settings but it is recommended that the user checks and corrects the system parameters 1 Switch on the printer 2 Switch on the PC 3 Start your terminal emulator 4 If necessary change the communication parameters in the terminal emulator 5 Insert the MicroBeta program disk into drive A the lower disk drive of the counter 6 Insert the MicroBeta protocol disk into drive B the upper disk drive of the counter 7 Switch on the counter with the power switch on the back of the counter 8 After about 2 minutes a message like the following will appear Incorrect terminal type GenTerm or MultiCalc VT52 or VT100 used Please select terminal type in S ystem Ter m inal 9 Press the S key and the System menu is displayed 10 Press the M key to change terminal parameters 11 Change to your terminal emulation by entering the number followed by Enter 12 If necessary change the terminal parameters as described in section 2J 30 10 System Terminal in this manual 13 Press any key e g the space bar when you are told to change emulator 291 4 1 Installation instructions 14 The program will return to the System state 15 If necessary change the RS communication parameters as described in section 2J 30 7 System RS 232C in this manual 16 Press the Q key 17 Ready is displayed
251. ses a special GenTerm WT emulation to direct output to the display the printer and the result file Result file saving is automatic and it is enough for you to only select on the Protocols File output line which data should be saved 2 31 4 9 Printer connection The MicroBeta program can automatically direct printer output through GenTerm to the printer connected to the terminal PC with a standard Centronics cable The printer is normally connected to port 3 on MicroBeta 2 31 4 10 Exit to MS DOS To get to the menu press Ctrl Break or Ctrl Scroll Lock 1f the Break key is missing Select either Exit to DOS temporarily temporary exit or Exit from GenTerm to DOS total exit press Enter and follow the instructions During counting it is recommended that you type D see chapter 2 16 Interrupt before temporary exit from GenTerm to MS DOS It is then also possible to exit totally from GenTerm and to use the terminal PC for running other programs After returning to GenTerm press D again to allow MicroBeta to send data to GenTerm See the GenTerm User Manual for details of other menu operations 2 31 4 11 Troubleshooting 1 Communication problems Communication problems e g characters are displayed correctly but the keyboard does not function will occur if MicroBeta is configured to use GenTerm WT emulation and the terminal emulator uses some other emulation UltroTerm VT52 or vice versa If you suspect that kind of situation then r
252. shake 1 None 2 DTR 3 XON XOFF 2 gt Set terminal port parameters Y N gt Y Are you sure Y N gt Y De you see this Gext Y N TY 2 30 8 P rinter Set the printer type and the number of characters per inch The options are IBM or Epson FX and the number of characters per inch 10 12 or 17 The defaults are Epson FX and 12 cpi Example 7 System gt P Printer type 1 IBM 2 EpsonEX 2 gt Printer charac per inch TOO eZ DNA A 2 30 9 D ata drive Select diskette or hard disk drive of the terminal PC or MicroBeta on which the results are to be stored when using UltroTerm define the result file path in UltroTerm Result filing options The result file path can also be given in each protocol Example 8 system gt D Data drive LRA 2 B 57 ROS 4 Path 4 gt Path name Cs MB RESUL TS gt The options are 1 diskette drive A Select this if you have a a single drive terminal PC no other possibilities or 150 2 30 System b a hard disk PC but you want to store results on the diskette e g to be transferred somewhere else 2 diskette drive B Select this if you have a dual diskette drive terminal PC Then terminal emulator software can be kept in drive A all the time e g to enable automatic restart after power failure 3 hard disk drive C Select this 1f your terminal PC is equipped with hard disk and you want large storage capacity If the GenTerm terminal emulator is used then results will be st
253. specific directory or d on the MicroBeta drive If 4 1s selected then the program asks for the full drive and directory path Here understanding of MS DOS drive and directory concepts is needed The directory path given here will be used as such without any checking and modifications in front of the file name Note that the directories must exist before any data can be sent to them Use the command MKDIR in DOS to make new directories Examples referring to examples a d above a Path name D gt Text D will be inserted in front of result file name E g D 2 003 the file will be stored in the active directory on drive D b Path name F gt Here it is supposed that drive F belongs to the LAN server Text F will be inserted in front of the result file name E g F 17 002 the file will be stored in the active directory of the LAN server drive F 237 2J 30 System c Path name C MB RESULTS gt Text C MB RESULTS will be inserted in front of result file name Note that if you specify an exact directory path for the result files then the path name must end with backslash For example the path c MB RESULTS 3 023 stores result file 3 023 into the directory MB RESULTS of hard disk drive C d Path name B DATA gt Results are stored on the MicroBeta protocol disk B in directory pata The character at the head of the path shows that the MicroBeta disk 1s to be used for storing the results
254. sponds to time t 0 described earlier in the formula This is the moment back to which all results are corrected If you select N to the zero time setting question then activities are corrected to the time when counting of the batch of samples was started Half life Half life hours 12 3 years 107500 5760 years eo 14 3 days 87 2 days 2093 27 8 days 60 0 days 1440 If you select a suitable half life correction then for example if a batch is recounted after e g one week the corrected CPM results are directly comparable with each other since they are half life corrected back to the same date and time N B Always check that the date and time are set to correct values in MicroBeta The current time can be seen in the upper right corner of the screen during counting If these values are wrong set them in the System state see Clock 72 2 15 Help and Info 2 15 Help and Info 2 15 1 MicroBeta TriLux Help The MicroBeta program has a Help function This means that information can be displayed to explain what options are open to you at the phase of operation you are in This information can be obtained by pressing one of the keys H h or In protocol editing press Enter also These help keys can be pressed at any stage of the program with one exception The H keys must not be used to get help when H has another meaning e g when defining active rows in the protocol editor Use to get help at that point 2 15 2 MultiCalc Help
255. ssette Press Y Yes if you want to count the whole cassette default or N No if you want to specify the positions to be counted If No has been selected the next parameter will appear 21 Cassette type Type if you use cassettes with 96 wells default or 2 if the cassettes have 24 or 384 wells 116 2 25 Protocols 22 Active rows If you answered No on line 20 you will have to specify the rows to be counted for the first cassette by typing the letters for these rows E g AC means that samples Al to A12 and Cl to C12 will be counted For a more detailed specification 1 e giving the positions instead of rows only press R Row coding In this case specify which positions are to be counted for each row The positions are in order from 1 to 12 or 6 for 24 well plates Type 1 to count a position or O to skip it E g Row A gt 0011110 means that positions A3 A4 AS and A6 should be counted You do not have to enter all the zeros at the end of the line the row end 1s filled with O or 1 according to the last typed character See the following example 20 Count all positions in cassette Y N Y gt N 21 Cassette type 1 96 2 24 gt gt Ze Cassette 1 96 Rows AB gt R Row As LLITLLILLITL gt 00L TULIO Row B DULTT1T1000000 gt 0O0DO0LLO Row C3 00001 LO00C00 gt 0 Row D 000000000000 Cassette 2 96 Rows ABCDEFGH gt AB Cassette 3 96 Rows ABCDEFGH gt An alternative method is to type the column numbers e
256. st level To get this go to System by pressing S select Level by pressing L this does not appear on the computer display but the figure on the next page shows it Then select the Test level by pressing T Go back to the Ready state by pressing Q and there select V see the example below Then press T and enter the number of runs The instrument will run the cassettes once and print the results After this it only prints ID read errors Keep the door open and check that the instrument is running alright mechanically First cassette AE QS SS l elie NOTE Keep your fingers away from the cassette rack when the door is open 293 4 1 Installation instructions Ready gt V H elp O ult GOs ETE C lear conveyor Rack u p Rack d own Remove ca s s N ext cass P rev cass Cu r rent cass Ne x t pos G oto pos Si z e checkpoint C l ose detector Op e n detector Next le v el Reset r a ck I nit mask Change m ask Calib 0 pos Calib det bloc k J ust read IDs T est IDs F ocus ID reader Print ever y ID Print ra w IDs Print b ad IDs Park det Shelf 1 16 Calib sensor Di s pin Di s pOut Conveyor shelf 1 gt T Number or runs 100 gt 5 Assay 1 ELO 2 Cass 3 Func NORM she Lies aL Assay 4 Prous Cass 6 Func STD Shelf 4 Assay 7 Prous Cass 9 Funes STOP She lt s 11 Assay 10 Prot 11 Cass 12 Pune NOID Shelf 16 Conveyor stopped ID test results RUNS S Errors W Read once 20
257. t Edit or create and edit a protocol nn prot no Edit a protocol by giving protocol number D elete Delete a protocol S how Show protocol parameters Print Print a protocol L ist Print list of protocols De fault Give default values to a protocol C opy Copy one protocol to another Operations in the CPM Normalization Protocol State The same operations as in the Counting protocol state plus N ormalization data Print normalization data Operations in the DPM Standardization Protocol State The same operations as in the Counting protocol state plus R eplot Plot DPM standardization curve Operations in the MultiCalc Assay Protocol State Contains only operations S how P rint and L ist Operations in the Conveyor State Help Display Help text Q uit Back to the Ready state O ff Stop conveyor C lear conveyor Move cassette to rack and move rack to the middle Rack u p Move the rack up one level Rack d own Move the rack down one level Test level contains also Remove ca s s Move cassette to rack N ext cass Move next cassette to conveyor and read ID P rev cass Move previous cassette to conveyor and read ID Cu r rent cass Move current cassette to conveyor and read ID Ne x t pos Move cassette to the next position G oto pos Move cassette to a specified position C ose detector Close detector block Op e n detector Open detector block 194 Next le v el Res
258. t nn count prot no Dnit D ispense Nnn norm prot no a N Ip Dnn std prot no W ash D Suck b ack s A utomatic counting Co m mand yee Operate con v eyor S tatus Hel P y Quit Dotal count rate na M annual counting ext pos MS ROS Injector X L G oto pos E rror beep C lear conveyor S tatus display Disp e nser IN OUT V ersion C lock Serial n o Protocols gt Counting protocol gt Temp cali b Help Help HV On Off Qhuit Quit Detector u sage C ounting protocol E dit Se t up mode CPM n ormalization prot nn prot no R S 232C DPM s tandardization prof D elete P rinter M ultiCalc assay protocol S how D ata drive P rint Ter m inal L ist P a sswords De f ault Customi z ing C opy Detector temp DPM stand protocol gt Help MicroBeta A gt Q uit MB Back to Ready gt E dit nn prot no D elete MultiCalc assay protocol gt S how H elp P rint Q uit L ist S how De f ault P rint C opy L ist R eplot 299 CPM norm protocol gt H elp Q uit E dit nn prot no D elete S how P rint L ist De f ault C opy N ormalization data Conveyor gt H elp Q uit O ff C lear conveyor Rack u p Rack d own Remove ca s s N ext cass P rev cass Cu r ent cass Ne x t pos G oto pos C Dose detector Op e n detector Next le v el Reset r a ck Dnit mask Change m ask T est IDs F ocus ID reader J ust read IDs Print
259. t 2 Operating information This part has information common to both MicroBeta TriLux and MicroBeta JET and also information only valid for MicroBeta TriLux In the latter case there are parallel chapters at the end of this part which are valid only for MicroBeta JET These chapters are given the same number as the MicroBeta TriLux chapter but with a J after the 2 e g 2J 1 1s the chapter giving MicroBeta JET information corresponding to the MicroBeta TriLux information in chapter 2 1 There is information at the head of each MicroBeta TriLux chapter referring you to the relevant MicroBeta JET chapter 13 14 2 1 Beginning operation of MicroBeta TriLux 2 1 Beginning operation of MicroBeta TriLux See chapter 2J 1 for JET information The following conventions are followed all through the MicroBeta user interface 2 1 1 Menu selections Menu selections can be entered when the system is in one of the following states Ready gt Ready for next operation Provtocols gt Protocol operations Count gt Counting operations INTO Additional info System gt System level operations Counting protocol CPM normaliz protocol gt DPM standard protocol gt Assay protocol gt Conveyor gt TOt aL count rate gt Counting protocol editor CPM norm protocol editor DPM SCs PrOococol editor MultiCalc assay protocol operation Conveyor operation Total count rate manipulation Command options available e g those shown below are displayed
260. t all positions in cassette 21 Cassette type 41 Counting control The following parameters are new or changed when compared with the ones in the counting protocol 3 Crosstalk correction Press Y to use crosstalk correction otherwise select N see chapter 2 7 Crosstalk correction for further information 123 2 25 Protocols 10 Counting time for background The counting time for the background and crosstalk counting samples in crosstalk counting is given in seconds The minimum value is 0 1 and the maximum is 999 999 9 seconds The default value is 60 0 11 Counting time for standards The counting time for the standard samples is also given in seconds from 1 to 999 999 9 with 60 0s as the default value 41 Isotope activity setting Press Y to set activities for normalization samples This is isotope dependent and given in DPM The range is from 1000 0 to 999 999 9 The default value is 200 000 for isotope and 100 000 for isotope 2 If activities are given absolute normalization is done with efficiencies as count rates divided by the given activity If N is selected relative normalization is done with efficiencies as a fraction of the highest detector count rate 47 Strip plate Press Y if you use plates where crosstalk is different in the X and Y directions The crosstalk sample in G10 is counted in this case See chapter 2 23 Normalization for further information 2 25 9 Parameters in a standardization prot
261. t diskette is missing or is unformatted etc then GenTerm displays a Drive not ready window Put a properly formatted diskette into the correct drive and press R to continue result saving If you cannot solve the problem then press A to stop saving data 164 2 31 Terminal emulators 2 01 5 UltroTerm 2 31 5 1 Product number and manual 1221 244 UltroTerm 2 Terminal Emulator 1221 922 UltroTerm 2 User Manual 2 31 5 2 Version V2 0 or later 2 31 5 3 Installation a For details see the UltroTerm 2 User Manual chapter 3 Installation b Floppy disk installation follow steps 1 8 as described in the manual When you reach step 9 instead type INSTALLF 1205 and press Enter Note MicroBeta uses the same mode as Wallac 1205 Betaplate The number 1205 is therefore also used with MicroBeta The number can be changed in communication parameter setting c Hard disk installation follow steps 1 2 and 4 7 as described in the manual in step 3 instead type INSTALLH 1205 and press Enter d If installation was made on hard disk then select the way in which UltroTerm should be started supposing UltroTerm was installed into directory Cc UTERM 1 If UltroTerm should start automatically after the terminal PC start up then copy lines of C UTERM START BAT to the file C AUTOEXEC BAT e g using the COPY command COPY C AUTOEXEC BAT C UTERM START BAT C AUIOEXEC BAT 2 Execute UltroTerm always from directory C UTERM 3 Insert t
262. t is available MicroBeta software can produce statistical output values in one of the following situations a Value of Repeats parameter protocol line 41a 1s greater than 1 and less than or equal to 3 If Repeats is greater than 5 or if 384 well plates are used then statistical output is not produced b Value of Replicates parameter protocol line 41b is greater than 1 c Both a and b hold If the user does not specify statistical output values and format then default statistical output format will be used In this format a table will be output after the last repeat or after the replicate group The columns of the table are shown below the symbol used in programmable output is shown in parenthesis In this table stands for 1 or 2 N MEAN THEOR ERROR 5 OBS ERROR CHI SQUARED PROBABILITY 35 Mean value of CCPMz DPMz or LCPS MEANZ Theoretical standard error of mean as percentages TSEMz Standard error of mean as a percentage SEMz Reduced Chi squared for mean CHISQz Probability as a percentage Pz See the chapter 3 3 Calculation methods for exact definitions The first row of the statistics table shows values for CCPM1 DPM1 or LCPS and the second row only in the case of a double label assay shows values for CCPM2 or DPM2 See the printer output of the example in 2 29 3 The default for the above values will be output at the 2 sigma level If the user uses Programmable output selection and includ
263. t plate Cassette field in ID support plate Corrected count rate counts per minute Theoretical percentage error for CCPM Corrected count rate counts per minute for isotope 1 i Theoretical percentage error for CCPM1 Corrected count rate counts per minute for isotope 2 i Theoretical percentage error for CCPM2 i Corrected count rate counts per minute for window 3 Theoretical percentage error for CCPM3 i Corrected count rate counts per second for isotope 1 i Corrected count rate counts per second for isotope 2 gt Corrected count rate counts per second for window 3 Colour Graphics Adapter Channel Reduced chi square value for CCPM1 DPM1 or LCPS Reduced chi square value for CCPM2 or DPM2 Chemiluminescence in percentages in window E Cassette order number COUNTS1 Counts for isotope 1 COUNTS2 Counts for isotope 2 COUNTS3 Counts for window 3 2171 3 5 Abbreviations and acronyms used cpi CPM CPM1 CPM1 CPM2 CPM2 CPM3 CPM3 CPM_C CPM_NC Characters per inch Count rate counts per minute Count rate counts per minute for isotope 1 Theoretical percentage error for CPM Count rate counts per minute for isotope 2 i Theoretical percentage error for CPM2 Count rate counts per minute for window 3 i Theoretical percentage error for CPM3 E Coincidence count rate counts per minute Non coincidence count rate counts per minute CPM_AQP AQP count rate counts per minute CPSI CP
264. t rate correction Background corrected count rate r is calculated using the equation r 1 b 3 where 1 is from equation 1 and b is the normalization background count rate for the same detector and counting window ParaLux high efficiency backgrounds are subtracted first Efficiencies e mn are calculated using the equation 259 3 3 Calculation methods e mn r mn j 4 where r mn is isotope n background and half life corrected count rate in window m and jJ is the given normalization activity or the maximum background corrected count rate of isotope n in window 1 single label or in window 5 1024 dual label or the average count rate luminescence Efficiency percentage error is calculated using the equation de mn e mn V Si mn r mn Sb mny r mn 8 17 5 If j is the given activity then oj 0 3 3 3 Corrected count rate Corrected count rate headings CCPM CCPM1 CCPM is calculated in three steps in the following order Background correction Detector efficiency correction or crosstalk correction Half life correction The first two corrections also affect the theoretical error headings CCPM CCPM1 CCPM2 of the corrected count rate 3 3 3 1 Background correction If background sample is specified and measured or background is given in the protocol then count rate 1 is corrected for background using equation 3 r 1 b ParaLux high efficiency backgrounds are subtracted first If backg
265. t the data to be stored in When you have done this then return to the terminal emulator program by typing EXIT and pressing Enter 49 2 8 Datafiles The disk drive and the directory for the results files can also be selected in the protocol together with the output file selection This will override the selection made in the System state and will make it possible for users to have their own results files directories When you have selected the directory you can instruct the program to save data on it as follows 2 8 3 Saving data in datafiles Select protocol operations by typing P N or S when in the Protocols state P is for counting protocol N for normalization protocol and S for DPM standardization protocol Next choose the protocol to be edited by giving the group number and pressing Enter You can edit any parameters in the protocol but in particular go to line 36 File output and select which data you want to save see chapter 2 22 Protocols for details The example shows that the Long output has been selected Then on the line File path enter the disk drive and directory or leave it as it is if the drive defined in the System Data drive shell is used In this example a subdirectory TEST belonging to directory MB on PC disk A has been selected 36 File output LATO ZV SHOE 3 Long 4 Programmable it SS File path C MB gt A MB TEST File name extension 1 gt 40 Change special features Y N N gt Before starting coun
266. te MEAN1 MEAN2 outputs a mean value under the CCPM1 CCPM2 of the last replicate etc 145 2 29 Statistics POS A01 A02 A03 CTLME CCPM1 MEAN1 FES LIRA cee LID 229 SZ 502 MEAN 30699 4 CCPMIS TSEM1 2 2 2 al a9 8 ad sG CSENWA SEM1 MEAN2 112834 IO fod OE o Sho ASE Ll 146 CCPM23 TSEM2 ee ore U UT SEM2 2 30 System 2 30 System See chapter 2J 30 for JET information 2 30 1 Selecting System System settings control the general way MicroBeta works its defaults hardware connections etc To enter the System state press S in the Ready state You are then given a number of options as example shows These are described in more detail here Example 1 H elp I nfo C ount P rotocols S ystem Ready gt s TULE MS D O S tatus display V ersion VORrZ52zC P rinter r m inal P a sswords velp rrorbeep FLOCK ata drive T ustomi zZ ng E system gt 2 30 2 H elp Press H to get help about the various selections 2 30 3 E rror beep User or instrument error normally causes a beep This can be switched off by selecting N Example 2 System gt E Error be p Y N Y gt N 2 30 4 S tatus display Normally when counting is occurring its status is shown on the Terminal PC screen see chapter 2 5 Counting for more details The counting status display can be switched off or only currently counted positions can be shown Example
267. te AQP I and High efficiency counts can also be optimized 115 2 25 Protocols For PMT use select normal coincidence counting or use only upper or lower photomultiplier tubes Upper is the default if luminescence counting is selected 6 Isotope 2 Displayed only in dual label counting The higher energy isotope is selected The alternatives are the same as on line 5 with C as default isotope 7 Window 1 Displayed only if Other has been selected as isotope Enter the limits for window l ina range from 1 to 1024 8 Window 2 Displayed only if Other has been selected as isotope 2 Enter the limits for window 2 in a range from to 1024 9 Window 3 Displayed only if Other has been selected as isotope or isotope 2 in dual label counting Enter the limits for window 3 in a range from 1 to 1024 or Q 0 if no third window is used 10 Counting time The counting time is given in seconds The minimum value is 0 1 and the maximum is 999 999 9 seconds The default value is 60 0 If quick view is selected then the maximum plate time is requested This is the total counting time of the plate without the sample change time The default value is 960 0 12 Precision 2 sigma Counting stops when the standard deviation is below the two sigma value in all detectors Precision is given in percent in a range from 0 to 99 9 O means that no precision limit is used The default value is 0 2 20 Count all positions in ca
268. ted FLAG eget Normal termination time reached Normal termination time reached Po PREC E Precision reached MAX Hot sample more than 50 000 counts channel o INT Counting interrupted by user 94 2 22 MultiCalc operation 2 22 MultiCalc operation 2 22 1 Introduction MultiCalc is developed from RiaCalc the famous immunodiagnostic software package with nearly 10 000 users in hospitals and laboratories all over the world In contrast to RiaCalc the MultiCalc software can be customized to different labelling techniques it can handle assays in both sequential and plate format and it supports PC networks MultiCalc allows result evaluation screening microtitration plate format calculations with free replicate placing a wide range of quality control operations mainframe communication etc MultiCalc runs on any IBM PC or compatible Graphics supported are CGA EGA MCGA and VGA both colour and b w The contact between MultiCalc and MicroBeta is by means of a special communication protocol MultiCalc is operated by softkeys It assigns different functions to the ten function keys F1 to F10 according to the operating level it is on The starting set of softkey functions is called the Main menu and is what you get when MultiCalc has been first started up To get to the main menu from any other level you have to press the F9 key one or more times plus answer any questions about exiting and saving that the program may ask you on
269. ted CPS values 100 3 1 8 MicroBeta JET features In addition to the MicroBeta TriLux features described above MicroBeta JET makes it possible to measure flash luminescence dual luminescence and do kinetic studies because it allows reagent addition and mixing by dispensing MicroBeta JET can be purchased without the injector assembly or injector modules and these can be added later in the field by the user when required Also additional injector modules can be added An injector module is a pump with 1 2 3 or 6 syringes each feeding tubing that goes to the mask adapter which positions the tips of the tubes above the well s Each syringe and hence piece of tubing constitutes one channel A single detector instrument can have up to four modules each with one channel A multiple detector instrument can have up to two modules each with as many channels as there are detectors in the instrument 1 e 2 3 or 6 Each module and channel can be switched on or off independently The use of at least two injectors for the same well means that dual luminescence assays can be performed If reagent injection is selected in any particular counting protocol the mask adapter injector assembly is driven into position above the microplate The injector tips which are embedded in the mask adapter are automatically positioned over the wells to allow injection under optimum conditions The light resulting from the reaction passes through the fibre optic co
270. ter the File output This over rides any path set in System mode The drive and path defined in the System state is the default value see chapter 2J 30 9 System Data drive 224 2J 25 Protocols This is the terminal PC path If results should be stored on the MicroBeta disk then the path must begin with an O e g B DATA Ifa VT52 or VT100 terminal is used then results are stored on the protocol disk as default directory B RESULTS see 2 30 10 Terminal The file name extension the part after the period can be changed on the next line This must be a number in the range 1 to 999 Note this line is not available if UltroTerm is used 40 Change special features Press Y to change special features These are counting control background half life and chemiluminescence correction password and special plate type 41 Counting control This and the following questions will be displayed only 1f Y has been selected on the previous line Press Y to change the counting control parameters These are number of repeats replicates and cycles If N is selected then these parameters have no effect The number of repeats is the number of times each sample will be counted The minimum value is default and the maximum is 999 If repeat is greater than 5 results are not sorted and statistical results are not printed Repeats cannot be selected in crosstalk The number of replicates specifies how many replicates there are for a
271. ter to get the next page of protocols When you have found the one you want to look at type the number of that protocol and press Enter Type L List to printout the protocol list and P Print followed by protocol number and Enter to print a particular protocol 2J 25 3 Creating and editing protocols To create a new protocol with default values press E Edit Choose a protocol number not in the list and press Enter The first free protocol number is shown in the menu Protocols have already some values in them either default or old parameters If you want to set default parameters press F deFault and select the protocol number You will be asked for a line number when starting editing a protocol Press Enter to start editing from the beginning of the protocol or type a line number followed by Enter to edit a certain protocol line You can always jump between protocol lines by pressing L followed by a line number and Enter Type P and press Enter to jump to the previous line The up and down arrow keys can also be used The value in front of the arrow on each line is the default value Just press Enter if you do not want to change this value or type a new value and press Enter if it must be changed The Ctrl B Ctrl key held down and the B key pressed and Ctrl N keys can be used to move within the input text the left and right arrow keys can also be used See 2J 1 4 Editing input data NOTE Unnecessary lines in a protocol can be hid
272. tes Any combination of the two types of plates accepted by your MicroBeta 96 and 24 or 96 and 384 can be used The type of cassette 384 96 or 24 position cassette is recognized automatically by the counter E Last cassette 2 17 2 Loading Section The cassette rack 1s in its middle position when being loaded Cassettes should be loaded into the rack with the sample plate upwards and the ID labels towards you The cassette to be counted first should be inserted on the bottom shelf numbered with 1 and the next cassettes on the following shelves Empty shelves between cassettes are allowed and the first cassette must not necessary be on shelf no 1 as long as there 1s no other cassette below it After loading close the door properly 2 17 3 Operation First cassette The movement of the cassette rack and cassettes position on the transportation system normally occurs automatically without any need for assistance from the user However manual operation 1s also possible Press O Operate conveyor in the Ready or Count state to get the cassette movement functions Press O Off to stop the cassette movement C Clear to clear the conveyor 1 e T1 E AN all dl Me Hl 2 17 Loading the cassette rack move a cassette from the transportation system to the rack and to move the rack to the middle position U Up and D Down to move the rack up and down and Q Quit to return to the Ready state
273. that at least once a month you run counting protocol number 99 the GLP Good Laboratory Practice protocol Use this protocol to count the normalization sample plate supplied with the instrument The background H and C samples are counted with all detectors so background level CPM values and SQP values can easily be compared Recommended limits A Eff Bkg Eff Bkg If the values obtained by running the GLP protocol are outside the recommended limits please contact your service support person 239 3 2 Routine maintenance 254 3 3 Calculation methods 3 3 Calculation methods 3 3 1 Count rate Count rate 1 in counts per minute headings CPM CPM1 CPM2 CPM3 is calculated using the equation i 60 c t 1 where c is counts in the counting window and t is dead time corrected counting time heading CTIME in seconds In ParaLux high efficiency mode CPM1 CPM_C CPM_NC where CPM_C is normal coincidence count rate and CPM_NC is non coincidence count rate between discriminator channel and 1024 Count rate a sigma percentage error headings CPM CPM1 CPM2 CPM3 is calculated using the equation o Si i a 100 Vc 2 Above and in what follows amu can be either 1 0 or 2 0 3 3 2 Normalization 3 3 2 1 Detector efficiency normalization Detector efficiency normalization corrects for differences in detector responses Efficiencies and efficiency errors of each detector are calculated and stored for coun
274. that your data has been transferred to another diskette or to some PC then answer N No In that case after starting counting the program confirms deletion before operation by asking Delete result files saved on protocol disk and you can answer N NO to reject deletion or Y Yes to accept deletion 2 30 10 5 Print through terminal Select here where to connect the printer Answer N No if you do not need to connect the printer to the terminal PC Connect instead the printer to serial port 3 of MicroBeta with serial cable 1221 502 Y Yes to connect the printer to the terminal PC In this case connect the printer to the parallel port of the terminal PC with a Centronics cable 1221 122 2 30 10 6 Exit terminal setting To exit terminal setting press slash and Enter on any line Otherwise after you have answered all the questions described above the program confirms that you accept the change If not press N or else press Y which causes the following message to appear Prepare your terminal ready for emulation change Press any key when ready Now you can make the preparations possibly required for the emulation change e g to switch from GenTerm to UltroTerm When ready press any key Then if the change succeeded the program returns to the System state However if the message Terminal type check failed appears see example 11 it means that MicroBeta has found that the terminal program does not use the same emulation as was selected M
275. the 20 circle the 11th circle from the left and the 5 circle The month is marked using the rightmost 12 circles the first circle corresponding to January the second February etc The row of 7 small circles on the right hand edge next to sample column A12 to H12 is used for sample identification The circles stand for the sequence 1 2 4 8 16 32 64 By cutting off appropriate combinations of circles the filtermat can be marked with a number in the range to 127 31 2 4 Coding cassettes and filtermats 2 4 6 Plate ID reader If the optional plate ID reader is installed then it is possible to read barcode IDs from plates The following barcodes are supported Codabar Code 39 Code 128 Interleaved 2 of 5 UPL and EAN The ID should be fixed to the upper part of the right side 32 2 5 Counting 2 5 Counting 2 5 1 Starting counting First load samples as described in chapter 2 17 Loading the cassette rack You can then either start counting as a terminal operation with a terminal terminal emulator or terminal emulation in MultiCalc or as a MultiCalc operation In terminal operation the counter program must be in the Ready or Count state before you give the instruction to start counting The options are as follows 2 5 1 1 Automatic counting terminal operation If your cassettes have protocol ID labels press A in the Ready or Count state The counting protocol to be used will be determined by the ID labels If you
276. the default mode you need not change printer type if the printer supports this emulation 4 1 8 4 Clock setting Instrument Manual See 2J 30 System C lock Check the date and time and set them to the current values if needed 4 1 8 5 Data drive Reference Manual See chapter 2J 30 System D ata drive Default Drive A Select the diskette hard disk drive and the directory for the MicroBeta result files as described in the Instrument manual Depending on the terminal emulation set in section 4 1 8 1 you must do the following 4 1 8 5 1 GenTerm No need for extra settings Note See chapter 2 31 Terminal emulators GenTerm Result files 4 1 8 5 2 UltroTerm UltroTerm itself takes care of result file saving Note See chapter 2 31 Terminal emulators UltroTerm Result files 4 1 8 5 3 MultiCalc MultiCalc itself takes care of result file saving 299 4 1 Installation instructions Note See chapter 2 22 MultiCalc and the MultiCalc User Manual chapter on files 4 1 8 5 4 Other terminal emulator programs or terminals As commercial terminal emulators differ a lot it is not possible to give any advice which covers all of them Note See the chapter 2 31 Terminal emulators VT52 or VT100 compatible terminals Result files 4 1 8 6 RS 232C interface protocol Instrument Manual See chapter 2J 30 System R S 232C Default DTR means electronic DTR handshake protocol Port 1 Terminal 9600 baud 8 data bits 2 stop bits
277. the plate should be in position You must give the number s of the injector module s to be dispensed and also the numbers of the channels to be used Then give the volume to be dispensed The units are microlitres The minimum volume is 10 uL which is also the default The maximum volume is set by the Dispense volume max parameter in which the sample volume in the well before dispensing 1s also taken into account For test purposes you can have the mask adapter out and put a waste tray or an empty plate under the injectors to catch the liquid 2J 0 10 Suck b ack This command is used to empty the tubing after you have completed dispensing The liquid will be sucked back into the liquid reservoir not only emptying the tubing but ensuring that no liquid is wasted Press B to begin this operation This operation should be performed before Wash is done 2J 0 11 W ash This command is used for cleaning the tubing after operation has finished and the tubing has already been emptied of liquid by the Suckback command Make sure the mask adaptor has been moved out to the injector chamber and a waste tray has been put under the injectors before beginning this operation The input tubing must be in a reservoir of water or if necessary the cleaning liquid recommended by the manufacturer of the liquid you have been dispensing Press W to begin this operation You must give the number s of the injector module s to be washed and also the numbers of the cha
278. there is no success then change the number of data bits usually 7 or 8 bits come into question and try different baud rates again Usually parity and handshake differences should not prevent normal manual terminal use Another method is to switch the counter off and on again 235 2J 30 System The correct parameters terminal type baud rate and data bits are set automatically after instrument restart If they differ from the current parameters then a text like the following 1s shown Incorrect terminal baud rate 9600 bps used Please set terminal RS 232C parameters in S ystem R S 232C When it seems that communication is working go to the MicroBeta System mode and check the serial port settings and make sure that they are identical with settings of the terminal Example 6 System gt R RS 232C port number 1 gt Baud rate 1 300 2 600 3 1200 4 2400 5 4800 6 9600 6 gt Parity 1 None 2 Odd 3 Even 1 gt Data bits 8 gt Stop DIES 2 Handshake 1 None 2 DTR 3 XON XOFF 2 gt Set terminal port parameters Y N gt Y Are you sure Y N gt Y Do you see this text Y N 7Y Note for serial port 3 the connection to the injector modules the parameters must be Baud rate 6 Parity 1 Data bits 8 Stop bits 2 and handshake 1 2J 30 8 P rinter Set the printer type and the number of characters per inch The options are IBM or Epson FX and the number of characters per inch 10 12 or 17 The defaults are Epson
279. thing will be used or linear interpolation if SPA is selected for the sample type 25 Curve fit method This line is displayed only if you have selected Yes on the previous line The methods for standard curve fitting are 1 Smoothing spline default 2 Interpolation spline 3 Linear interpolation default for SPA 4 Linear regression Select a method by typing the number and Enter If you have selected the first alternative smoothing spline you will be asked if the method is to include automatic smoothing Press Y default if so Otherwise press N and you will be asked for a smoothing parameter Enter a value between 0 0 no smoothing and 1000 0 maximum smoothing with 1 0 as default 26 Edit standard curve Press Y to edit the quench parameter efficiency and weight values for a standard curve or to delete curve points 41 Adjusted activities Press Y if you want to adjust the activities for standards otherwise the activity given on line 21 will be used for every standard After pressing Y you will be able to change activities for the same number of DPM standards as given on line 20 You can also change the activities for the two Easy DPM standards The range is from 1000 0 to 999 999 9 DPM The value given on line 21 1s default See chapter 2 12 DPM Standardization for further information 230 2J 25 Protocols 2J 2 Robotic loading interface The robotic loading interface is a special door adapter system that brings a
280. tical uncertainty in samples is expressed as the standard deviation or sigma value and is calculated by dividing number one by the square root of the counting value E g 10000 counts gives a one sigma value of 1 The 2 sigma value is two times the one sigma value or 2 for 10000 counts In MicroBeta precision is given as 2 sigma in a range from 0 to 99 9 Zero means that no precision test 1s used Make sure that the counting time is set long enough if precision is wanted E g two samples of 1000 and 10000 CPMs will take 10 min and 1 min respectively to achieve a 2 sigma value of 2 Precision is checked once a second The termination flag in results output will be set to 37 2 6 Counting control PREC if the precision has been reached The counting stops if the precision has been reached in all detectors 2 6 4 Number of repeats To check sample stability or to check instrument performance the same sample can be counted repeatedly up to 99 times If the repeat value is less than or equal to 5 then results are sorted and some statistical values are calculated and printed after the sample results 2 6 5 Number of replicates To evaluate sample preparation errors a number of replicates of a sample can be used The results for each sample are output separately After each group of replicate samples some statistical values will be sent to output The maximum number of replicates are 99 2 6 6 Number of cycles Al
281. ting be sure that you have inserted an empty formatted data diskette into the drive selected or if you have selected saving to hard disk drive C be sure that the current directory has been selected as described in the previous section If the file path is given then this directory does not need to be the current directory but it must exist Use the MKDIR command in MS DOS to make new directories Format new diskettes by the MS DOS FORMAT command When you have finished protocol editing store the protocol by pressing and begin counting by pressing A in the Ready or Count state Note Line 36 is not available if UltroTerm is used The filing system of UltroTerm should be used in this case 2 8 4 Accessing datafiles Result files that are stored on the data disk will be named so that the first part of each name the part before the point is the protocol number and the second part after the point is a 50 2 8 Datafiles running number indicating the assay execution order E g for protocol 5 the first saved file is named 5 001 the second 5 002 etc Other files have the format e g S5 001 for standardization files and N5 001 for normalization files The file name extension can be changed in the protocol To access a file on the data disk in the terminal PC exit from the terminal emulator program to MS DOS see the chapter Terminal emulators Exit to MS DOS 2 8 5 External datafiles Results can be sent to an external data collectio
282. tions as a random access stack which allows access to any cassette in the rack The cassette rack 1s driven by a stepper motor and its movement is controlled by electro optical sensors A transportation system moving cassettes in the x y direction selects one cassette from the cassette rack and takes it into the counting position and then returns the cassette to the cassette rack This transportation system is driven by two stepper motors and is controlled by electro optical sensors 3 4 8 Mask adapter Injector assembly A transportation system for moving the mask adpater in the x direction from the maintenance tray into the counting chamber under the upper detector block This transportation system is driven by one stepper motor and is controlled by electro optical sensors The mask adapter moves in the z direction with the upper detector block 3 4 9 Detector assembly 1 2 3 6 or 12 detectors each consisting of a pair of 19 mm PM tubes working in coincidence are mounted in two detector assemblies the upper detector assembly moving in the z direction PM tubes are Hamamatsu R1166 or equivalent Coincidence resolution time is 100 ns One to six detector assemblies are provided with circular apertures for 96 and 24 well sample plates The 12 detector assembly is provided with circular apertures for 96 well sample plates and square apertures for 384 well sample plates The apertures are automatically changed to correspond to the plate type
283. to use crosstalk correction otherwise select N see chapter 2 7 Crosstalk correction for further information 10 Counting time for background The counting time for the background and crosstalk counting samples in crosstalk counting is given in seconds The minimum value is 0 1 and the maximum is 999 999 9 seconds The default value is 60 0 11 Counting time for standards The counting time for the standard samples is also given in seconds from 1 to 999 999 9 with 60 0s as the default value 41 Isotope activity setting Press Y to set activities for normalization samples This is isotope dependent and given in DPM The range is from 1000 0 to 999 999 9 The default value is 200 000 for isotope 1 and 100 000 for isotope 2 If activities are given absolute normalization is done with efficiencies as count rates divided by the given activity If N is selected relative normalization is done with efficiencies as a fraction of the highest detector count rate 47 Strip plate Press Y if you use plates where crosstalk is different in the X and Y directions The crosstalk sample in G10 is counted in this case See chapter 2J 23 Normalization for further information 228 2J 25 Protocols 2J 25 9 Parameters in a standardization protocol The parameters are much the same as in the counting and normalization protocols with some exceptions JET features are not available with standardization but the mask adapter can still be used Inf
284. tomized MENU N Environment string name gt MENU MENU gt Environment string name gt 155 2 30 System 2 30 13 MS D O S Press O to get to the MicroBeta MS DOS You must confirm this twice as example 14 shows after which the text MicroBeta A gt appears Example 14 system gt O Exit to MS DOS Y N gt Y Are you sure Y N gt Y Type MB to start the program again MicroBeta A gt MB H elp I nfo C ount P rotocols S ystem Ready gt Type MB and press Enter to get back to the MicroBeta program Note Avoid going to MS DOS in routine work The main reasons to go there are to make a backup copy of the protocol or program diskette 2 30 14 Q uit Press Q to quit from the System state 2 30 15 Se t up mode If a system password is used then the system mode menu is quite short see example 15 Example 15 H elp O ult MS D 0 S E rror beep S tatus display V ersion C lock Se t up mode System I Password gt H elp O ult MS D 0 S E rror beep S tatus display V ersion Cy Lock R S5 2320 P rinter D ata drive Ter m inal P a Sswords Customi 1 zing system gt 156 2 30 System All the functions that can affect routine use are hidden The person responsible for the MicroBeta system can make the hidden functions visible if he or she knows the password Press T to get into the setup mode The program then asks for the password Type the password and press Ente
285. ts such as 1450 423 Nylon membrane No scintillator needs to be added to the filtermat allowing reprobing assays The filtermat in a sample bag is placed between the base plate and the cover plate having solid scintillator in sample positions Cassette 1450 130 This has 384 sample holes This is used when counting 384 well sample plates All the cassettes are made of white plastic with an optical surface in the sample holes 2 2 2 Use The sample plates are placed on top of the cassettes the filters between the plates the cassette can be labelled with an ID code This is described in chapter 2 4 Coding the cassette Cassettes are loaded into MicroBeta as described in chapter 2 17 Loading the cassette rack A cassette type is recognised automatically 1 e whether it is a 384 96 or 24 position cassette that is being counted See chapter 2 9 for information about which positions detectors are in when counting samples in different types of plates 2 2 3 Adapters and inserts Adapter 1450 108 These adapters are used when counting Eppendorf tubes or equivalent The adapters are place on a 1450 102 cassette a bottom tape is used if needed Inserts 1450 109 These inserts are used when counting 24 well culture plates The inserts are placed in the sample wells in order to reduce crosstalk between the wells 26 2 3 Clock settings 2 3 Clock setting 2 3 1 Current date and time The current date and time are shown in the upper right
286. twice Op thrice 0 The text above should be printed 1f the test is OK Check that there are not many occasions when the ID label is read twice or thrice because this shows that the ID reader is performing poorly 4 1 7 3 Test settings For checking the performance of the instrument run the detector efficiency normalization with the normalization standard plate supplied with the instrument Attach the printout of the measurement to the installation report 4 1 7 4 System operation showing the TEST level The figure on the next page is a flow diagram of the whole MicroBeta operating system It shows how the main control letters in parentheses lead to the various functions and how the test functions are related to the normal functions Compare with the flow diagram in 2J 1 5 showing the functions on the normal level 4 1 7 5 Leaving the TEST level To return to the normal level press Q to return to the Ready state then S to go to the System state Press L for level selection and N for normal level not shown Press Q to return to the Ready state 294 Flow chart of MicroBeta commands service level 4 1 Installation instructions nfo gt Hel F Total count rate gt Ready gt Qui Help Help U sage C bl Q uit I nfo C onsumables e S how C ount P ositions a Ch a nges P rint P rotocols BES R Cu s tomizing R eset S ystem a Define efficiencies Count gt Injector gt JET only Help Help Q uit Q ui
287. uch a plate Place the plate with the normalization samples in a cassette that is identified with the corresponding normalization protocol number in the protocol number area and with a NORM label in the function code area 205 2J 18 Luminescence counting Insert the cassette s into the rack and close the door Press either A Automatic counting or Nxx where xx 1s the normalization protocol number in the Ready or Count state to start counting If Nxx is selected the program asks Do you want to continue counting after normalization Y N gt If Y Yes is selected the counting continues after counting the normalization plate if N No is selected the counting stops after the normalization If normalization has already been done for the protocol then the program asks first This PpEo tocol Contains normalization data Continue Y N gt This is to prevent accidental overwriting of normalization data If automatic counting is selected the normalization cassette can be placed anywhere in the rack The NORM label is needed in this case The counting results for the normalization are output as specified on lines 30 37 in the normalization protocol and after them the background and detector efficiency values are given 2J 18 8 Counting luminescence samples Edit a counting protocol In the Ready state select P P rotocol then C C ounting protocols Then select edit E and protocol number On parameter line 2 select
288. umber of labels 1 Single 2 Dual Isotope 1 IHH AVERLO 3 G li4 4 S 35 De anor 6 P 32 OT aS Go Nor 7 P 32 Cerenkov 8 Other ParaLux Y N PMT use 1 Normal 2 Upper 3 Lower Window 1 6 Isotope 2 Le ZN SJ Cale Ay S 35 Sep 6 P 32 7 P 32 Cerenkov 8 Other Protocol name Dual CPM Window 2 150 650 10 Counting time s 12 Precision 2 sigma 20 Count all positions in cassette Y N 21 Cassette type 1 96 2 24 ZZ Cassette 1 96 Active rows Row Row Row Row Row Row Row Row Cassette 2 96 Active rows 23 Plate orientation 1 Normal 2 Rotated 3 Mirrored 4 Both 30 Printer Outp t 1 No 2 SROLE 3 LONG 4 Programmable 1203450000 y AB AAA AA 111100000000 000000000000 000000000000 000000000000 000000000000 000000000000 gt 000000000000 ACEG A 4 sl Printer OUT DUE TPOS CIIME SOP GL CEPML CCEPMIS gt CCPM2 CCPM2 PROT SPECTRA Plot energy scale 1 Logarithmic ch 2 Linear keV 3 Bothspectra af Plot area channels La Loo 4 Plot size 1 Small 2 Medium 3 Large 32 Display outputs 1 No 2 NOE 3 Long 4 Programmable 34 External output 1 2 Short 3 Long 4 Programmable 36 Fileoutput 1 2rSHOrE 113 2 3 1 2 25 Protocols 3 Long 4 Programmable 4 a File QUEUE POS CTIME SOP LL SOP 1 2 COUNTSL gt CCPM1 CCPM1 COUNTS2 CCPM2 gt CCPM2 FLAG STM PROT Pi Le patas CEXMBLASONTESTN File name extension iL 40 Change special features Y N Y Al Conntrai
289. upling in the injector assembly to the upper PMT 251 3 1 Instrument description The mask adapter can also be used without injection simply for masking It reduces the signal to the upper detector by a factor of ten which can be helpful in cases of luminescence with very high signal 3 1 9 Robotic loading interface The robotic loading interface is a special door adapter system that brings a cassette outside so that a robotic arm can change the plate In this case only one shelf is used Note the mask adapter has to be moved manually into the counting chamber before starting counting 3 1 10 Microcomputer control Operation of the instrument and calculation of the results is performed by a combination of a terminal PC running a Wallac terminal interface program or the MultiCalc immunoassay program and the instrument itself The built in computer is composed of a 16 bit Intel 80186 microprocessor with 1 MB RAM and 64 kB ROM There are two microfloppy 3 5 disk drives each disk has a capacity of 1 44 MB The disks loaded onto these drives contain the program and the protocols The Terminal PC is a personal computer that is used to run the terminal emulation program or MultiCalc and can be used to collect counting data for further evaluation Output possibilities can be selected by means of the programmable RS 232C interface with three output ports The uppermost port sends the output to the terminal the second to an external computer
290. urve 126 230 Editing input data 16 188 Efficiency 269 Absolute 103 207 Relative 103 207 Efficiency correction Detector efficiency correction 257 Efficiency normalization 255 Elapsed time 28 Electronics 268 End time 28 Energy calibration DPM standardization 63 Enter key 16 188 Error beep 147 233 Errors 69 EXCL 130 External computer Port 2 49 External computer Sending data to 49 External output 120 145 224 F Falcon plate 123 227 Faults 252 Fibre optics 251 Cleaning 253 File output 120 145 224 Filling wells 88 Filtermat 311 5 Index Coding 31 Filtermats 273 Flags 93 Foil heatsealing 275 Format Microtitration plate 83 Format command 50 FUNC Function codes 29 Function codes 29 G GenTerm 152 161 238 Changing parameters 162 Exit to MS DOS 163 Installation 161 Printer connection 163 Restrictions 163 Result files 163 Special features 162 Start up 161 Trouble shooting 163 GLP protocol 111 215 253 Good Laboratory Practice 111 215 253 Goto pos 186 H H Help 15 73 187 Half life 71 Half life correction 122 129 226 258 Handshaking 149 235 Help 15 73 187 System 147 233 Hiding text 112 216 High efficiency 115 219 255 High voltage 250 HiSafe 88 How to control MicroBeta 15 187 I Id label Fixing 29 ID label 253 ID products 275 ID support plate 29 5 Index ID system 268 Info 73 Init 184 Initia
291. used In dual label DPM counting three values are given The value is in the range from 0 0 default to 99 9 CPM or 999 999 9 LCPS Note in the normalization protocol press Y if you want to count the background plate 43 Half life correction Press Y 1f you want to use half life correction The half life for the isotope is displayed e g 107 500 hours for H No half life value is given for C because it is so long Note that you must always specify the half life if you have selected an isotope of type Other It must be in a range from 0 no half life to 999 999 99 hours 114 years If normalization or standardization protocol is selected on line 3 then half life is as specified in this protocol and it cannot be changed in the counting protocol Press Y on the next line if you want to set zero time otherwise the activity is corrected to the time when counting of the first sample of the assay was started The zero time consists of date and time First enter the date in the format DD MMM YYYY where DD is the day number 1 31 mum the month in letters JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV and DEC and yvvy the year e g 15 JUN 2000 The date can also be in the format yyYY MM DD e g 2000 06 15 Then enter clock time in the format HH MM SS where HH 1s hours in the 24 hour mode 0 23 mm minutes 0 59 and ss seconds 0 59 e g 14 05 30 It is not necessary to give seconds The default value for zero ti
292. vailable Positions View normalization and standardization positions Ch a nges View a list of changes after the previous version Cu s tomizing View information on how to customize the program S y stem View information about the system Operations in the Count State Help Display Help text Q uit Back to the Ready state nn count prot no Start automatic counting by giving protocol number Start automatic counting by selecting protocol from list Nnn norm prot no Start CPM normalization by giving protocol number N Start CPM normalization by selecting protocol from list Dnn std prot no Start DPM standardization by giving protocol number D Start DPM standardization by selecting protocol from list A utomatic counting Start automatic counting using IDs Operate con v eyor Conveyor operations T otal count rate Show and reset total counts M anual counting Start manual counting only at Test level Operations in the Protocols State H elp Display Help text Q uit Back to the Ready state C ounting protocol Edit counting protocol CPM n ormalization prot Edit CPM normalization protocol DPM s tandardization prot Edit DPM standardization protocol M ultiCalc assay protocol Show and print MultiCalc assay protocol 193 2J 1 Beginning operation of MicroBeta JET Operations in the Counting Protocol State H elp Display Help text Q uit Back to the Ready state E di
293. vel for all deviation output fields i Standard deviation for CCPM1 DPM1 or LCPS Standard deviation for CCPM2 or DPM2 i Standard error of mean for CCPM1 DPM1 or LCPS Percentage standard error of mean for CCPM1 DPM1 or LCPS i Standard error of mean for CCPM2 or DPM2 Percentage standard error of mean for CCPM2 or DPM2 Running sequential sample number Shelf number Scintillation proximity assay SPECTRA Plot spectra i SPSAVE SQP I SQPE SOPDm SQRT STAT STD STIME STM STOP TAB TCA TCV1 TCV2 TIME TSD1 TSD2 TSEM 1 TSEM1 TSEM2 TSEM2 TXT UN UTMAC VGA VT100 VT52 Wi W2 W3 Save spectra to file in compressed numerical format l Spectrum quench parameter of isotope spectrum i Theoretical percentage error for SQPA SQP for isotope m Square root Data specified will be output when a statistics mean is available Standardization ID label Counting start time Statistics monitor Stop ID label Produces tab character between all output columns Trichloroacetic acid Percentage theoretical coefficient of variation for CCPM1 DPM1 or LCPS Percentage theoretical coefficient of variation for CCPM2 or DPM2 Current time i Theoretical standard deviation for CCPM1 DPM1 or LCPS i Theoretical standard deviation for CCPM2 or DPM2 l Theoretical standard error of mean for CCPM1 DPM1 or LCPS i Percentage theoretical standard error of mean for CCPM1 DPM1 or LCPS E Theoretical standard error of mean for CC
294. volume of 750 microlitres high flash point cocktail unpurged in the 1450 402 24 well Sample Plate typically 24 CPM in wide H window typically 37 CPM in wide C window 3 4 13 2 Efficiency 1450 471 Normalization Standard H gt 45 1C gt 90 Unquenched sample with a volume of 150 microlitres high flash point cocktail unpurged in the 1450 401 96 well Sample Plate H typically 57 C typically 94 Unquenched sample with a volume of 750 microlitres high flash point cocktail unpurged in the 1450 402 24 well Sample Plate H typically 61 C typically 95 3 4 13 3 Crosstalk Optical crosstalk due to light piping in the plastic of the sample plates The values are given for sample plates with black printing inhibiting crosstalk between the sample wells Unquenched sample with a volume of 150 microlitres high flash point cocktail unpurged in the 1450 401 96 well Sample Plate In wide H window lt 0 05 In wide C window typically 1 8 Unquenched sample with a volume of 750 microlitres in the 1450 402 24 well Sample Plate 269 3 4 Specifications In wide H window lt 0 02 In wide C window typically 0 2 3 4 13 4 Stability Count variation less than 0 5 24 hours not including random statistics 3 4 14 Program Specifications 3 4 14 1 Counting protocols 100 fully programmable counting protocols Start of counting manually through keyboard command or automatically by the us
295. w kBa Hes 32597 139 oy eye 0 LISO 1087 LS 0 0 0 0 EC 14 124189482 90 0 2978592 62 151 2300 300 86669 0 O 0 Cea oi 0 0 0 0 P 32 0 0 0 0 P 32C 0 0 0 0 Other 2014554 0 0 0 H elp Q uit S how P rint R eset D efine efficiencies Total countrate gt 178 2 34 Window settings 2 34 Window settings 2 34 1 Radiation Energy Radioactive isotopes disintegrate spontaneously The disintegration results in emissions of alpha beta or gamma radiation The radiation type and the energy emitted is specific for each isotope In the case of beta radiation the emitted beta particles have a wide range of energies the energy spectrum forming a continuous distribution Beta radiation energy ranging from 0 to 2000 keV can be measured in MicroBeta the most common beta emitters have their energies between these energy values The energy scale is converted into a logarithmic scale and divided into 1024 channels The energy spectrum of a beta emitting isotope falls into a certain segment of the energy scale called an energy window A window is a part of the energy scale consisting of many adjacent channels e g a window beginning from channel 5 and extending to channel 360 will cover the energy spectrum of tritium in MicroBeta 2 34 2 Window setting The MicroBeta LSC has built in windows for the isotopes H la I a S PP a Cr 2p 32p Cerenkov and for Luminescence counting The window for Other can be set by the user The default value is 5
296. wed by Enter to edit a certain protocol line You can always jump between protocol lines by pressing L followed by a line number and Enter Type P and press Enter to jump to the previous line The up and down arrow keys can also be used The value in front of the arrow on each line is the default value Just press Enter if you do not want to change this value or type a new value and Enter if it must be changed The Ctrl B Ctrl key held down and the B key pressed and Ctrl N keys can be used to move within the input text the left and right arrow keys can also be used See 2 1 4 Editing input data NOTE Unnecessary lines in a protocol can be hidden to make the editor shorter Press Ctrl K Ctrl and K keys pressed at the same time to hide the current line To reveal a hidden line go to the next line and press Ctrl U To unhide all hidden lines go to line selection line in the beginning of the editor and press Ctrl A Press and Enter when you are finished with your editing and want to save the changes made If you want to exit without saving the new values press and Enter 112 2 25 4 Example protocol listing 2 25 Protocols The example alongside shows a typical dual label protocol listing The parameters are described in section 2 25 7 Counting protocol no 26 Mon 24 May 2000 10 49 Name Dual CPM CPM normalization protocol no Wallac 1450 MicroBeta V4 4 S N 4500759 Counting mode 1 CPM 2 DPM CPM normalization protocol N
297. y fuse in its holder into the socket at the rear of the instrument 4 1 5 Connecting up the counter and peripherals Connect the counter to the PC and the printer using the cables shown in the figure MicroBeta TnLux connections Printer connected to PC for woprkstation and Multi alec operation Cable 224 122 Printer connected ta MicroBeta for output of terminal operation Cable Printer connected to PC for workstation and Multic alec operation Cable 1224 122 Cable lt i 1221 503 289 4 1 Installation instructions There are two configurations either the printer and PC are each connected to MicroBeta to ports 3 and 1 respectively not possible with MicroBeta JET or the printer is connected to the PC which is in turn connected to port 1 This latter configuration is for use with the Windows workstation and MultiCalc See the additional section at the end of the chapter for information about Installation of the Injector system to MicroBeta JET Plug in the power cables for each device 4 1 6 Start up Although the UltroTerm terminal emulator program is the default emulator for MicroBeta other emulators e g GenTerm and MultiCalc or VT52 and VT100 terminals can also be used The following two sections describe how to start MicroBeta with the default UltroTerm and with another emulator 4 1 6 1 Start up using UltroTerm This is the default setting and therefore no parameters must usually be c
298. y protocol gt Counting protocol editor CPM norm Protocol edito DPM std Protocol editor MultiCalc assay protocol operation Conveyor gt Conveyor operation Total count rate gt Total count rate manipulation Injector gt Injector operations Command options available e g those shown below are displayed before the prompt text e g Ready H elp I nfo C ount P rotocols S ystem Ready gt The command is executed by pressing the letter either lower or upper case enclosed inside parentheses However commands which include a protocol number e g giving a pure protocol number in Ready state to start counting of that protocol must be completed by pressing the Enter key Help text can be obtained by pressing H h or Exit to a higher level by pressing Q Pressing Control E the Ctrl and the E keys at the same time exits always to the Ready state 2J 1 2 Confirming questions and messages Quite often MicroBeta displays a message such as Press any key when ready or to exit gt This kind of message is used to give to the user time to think 1f this is really the correct operation and to check that everything is prepared for the operation Press any key such as space bar when ready to continue Otherwise the user can cancel the operation by pressing slash 187 2J 1 Beginning operation of MicroBeta JET Messages such as Do you accept the new terminal setting Y N gt are used to notify the user that t
299. zation counting Start the normalization run in the Ready or Count state using automatic counting and ID codes or by selecting Nx where x is the number of the protocol 2 18 6 Counting the actual samples Edit a counting protocol In the Ready state select P P rotocol then C C ounting protocols Then select edit E and protocol number On parameter line 2 select 1 CPM and on line 3 select the number of the luminescence normalization protocol defined above On line 42 select Y Background correction and on the next line Y Use normalization background Edit the other parameters normally Start counting in the Ready or Count state using automatic counting and a protocol ID label or start with the protocol number and press Enter 81 2 18 Luminescence counting When counting the samples the CPS values are shown on the status display The stored normalization factors are used for correcting the LCPS values in the printout The normalized LCPS values are shown after counting on the status display 82 2 19 Microtitration plate format output 2 19 Microtitration plate format output 2 19 1 Advantages of plate format It is often more natural to print out MicroBeta counting results in microtitration plate format because then it is easier to locate the results for a given sample Plate format also has the advantage that it often reduces the paper consumption in printer output 2 19 2 Plate format selection Microtitration plat

Instrument Manual

Contents

Download Pdf Manuals

Related Search

Related Contents