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Gamma Counter 1470 Wizard

1. seeseee 64 Lave diSpldy ise eet aiii Load protocol essere Extended load Normal load Low high boundary M Main frame i iiie etre ete ite aro Mains voltage selection Manual background normalization 91 Manual GLP normalization oocccnonoccnononccnoonnnononaninonanoss 92 Manual normalization Manual operation cessere Many DeakS iens eteneciee oet certes eee eene ER eREe Marker Max counts limit Maximum assay deviation 96 sss Maximum coincidence deviation 96 Maximum detector efficiency deviation 96 Maximum normalization deviation ssss 96 MCA etit Ea deidad 127 MCA high limit esee 95 Menu SAM ine nasse ee 19 Microspheres u en anne 120 Minimum coincidence height 96 Mode Manual or automatic caeca eoe o HERE EH 89 Mode selection Monito iino iota pp HO ECHTE MultiCalc Mode lada 89 Real timeclock nd 70 Technology selecto cri ck 185 MultiCalc installation Multicalc OperatiON ooonoonnocincnnconocconnonnnonnconcnononccnnnco nono MultiCalc protocol operations eeeeeee 68 Multichannel analyzer esses 121 N No dynamic normalization sees 98 Nominal gain senden ana 102 Norm printout full
2. sss 104 Print paper length cin ede 100 Print protocol zu e ett ette acti 32 41 undc P oK 45 Printer ty Pe cei teneret ene ren tata 100 104 Printout field Deleg cucine lini eC 50 Diog cc 50 Pastine acut Helms iie iei eie e iubes 51 Printout fields Printout options sess 49 99 Printout selections sess 100 Printout switches PROTOCOL ate 2 nein 24 Protocol definition sse 65 Protocol editing Protocol labels nennen ten Prtotocol type erae ee tret tine she iberia adas Purge protocol eeeseeeeeeeeeneen nene 13 Index Q Quality control iaa iie teint tien head lies 67 R Rack ID reader selection sess 102 Rack number labels RACK SPECIAL Racks DMI 125 Loading tight way round venis lidia 23 Read spectra from diskette sss 104 Real time cl ck eee nnt casero no terrenus 70 Recover protocol 31 41 Rename protocol 31 41 Repeat counting eene 80 Repeat remainder of samples sssssss 47 Repeat times 94 Ii e C aE 45 Results Ale 45 Running an assay m Running an assay with MultiCalc 68 Sample ID Sample rayo mee e OIM RIPE 89 Save protocol Normal saves eie aci reti te e edere 42 Save Protocol arrsa eannan asns nennen 32 DAVE Protocol E 42 Exte
3. eee 99 Normalization essere 120 Background manual 91 Dual label sali GLP manual 1 92 GEM re 79 e A rel 78 Label Manual ING GynDamie ani a 98 Preset isotopes 2 PFIDECIBIG iia iu icto d I p oW EE boda porta 7 Printout un u ir 78 Rack for GLP 19 197 13 Index Operating the CONVeyYOT ooccnnccocononconnnononononnconocnnonnnnnncnnos 18 Operation Modena iia euere 97 O OIRLO T AETA A 47 Output IT mnan ienn a S 103 Output device status essere 105 Output editing i ret reri ile 48 Output from RiaCale WIZ sse 59 Output media doo ee need 48 Output negative CPM as 0 101 Output port selection uneeenssennesennennesnnnn nennen nenn 104 Outputs from MultiCalc sse 73 P Pagination of output 45 Paper length eet et rt i rre Re e eire 100 Parameters with CPM operation 33 Parameters with RiaCalc WIZ PC keyboard Peak Positioune niisiis Peak positions sese Port selection iss ini asian Ports Communication parameters eese 105 Error messages Power failures E ann Print Isotope DIIS variacion iria 42 Print dead time factor see 103 Print diagnostic info essere 103 Print isotope and normalization data 97 Print measurement start time esses 103 Print model and version
4. 1470 Main Menu OPERATE UNIX FILES SYSTEM Submenu Create Copy Rename Delete Recover Print Load RiaCale WIZ Press START to measure Create Select CREATE and press E Give the protocol name There is a default name of the form Protnn where nn is the protocol number If you want to change this name you must use the external keyboard unless you only use numbers for the name A maximum of eight characters can be given Select an unused ID number between to 99 Give the assay type RIA or IRMA If you just want to get CPM results or you do screening select RATIO Press E The protocol parameters can be set in a similar way to those in editing see below Edit Select Edit and press E Enter Select the protocol by name from the list of protocols available and press E Edit protocol 05 BF BLAN 06 FOLATE 07 FORMAT 08 LEARN 09 PROTOI 10 PROTO2 11 PROTO3 12 READ ME 13 PROTOO 14 TEST_1 15 TEST_3 16 TSH Id order change with lt gt 40 4 Operation with internal RiaCalc WIZ You may now change the parameters You can move up and down in the parameter list by means of the UP and DOWN arrow keys See section 4 5 for details of parameters available in a protocol Edit protocol 11 PROT 03 Dual evaluation NO Counting time 60 Max counts limit 9999999 Labels 1 125 X axis conc LOG Y axis resp B BO Fitting algorithm AuSpline Std outlier reject NO Controls NO
5. 10 1 Background normalization 10 1 5 Calculation of background CPM in the isotope counting window Counts are summed over the isotope counting window the result is multiplied by the dead time correction factor and converted to CPM In the printout these CPM values are further divided by the relative detector efficiency 1470 only However the background values stored internally do not have this correction Therefore in assay counting background activity is subtracted before correction for relative detector efficiency is made As far as assay counting is concerned background and isotope normalization can be made in any order Background normalization uses the latest effective gain value to calculate background CPM in all isotope counting windows and updates isotope normalization results with the measured background activity During isotope normalization the latest background spectra are used to update background CPM to correspond to the latest counting windows However when crosstalk and spillover correction coefficients are calculated the latest background normalization done prior to the isotope normalization is used This is a reason to do background normalization before isotope normalization s When the program determines peak positions in terms of MCA channels it uses interpolation to estimate the peak position as a fraction of channel Likewise counting window boundaries are determined to a fraction of channel When counts are summed over
6. 7 2 7 Manual normalization All operations which are available in automatic operation are also available in manual mode these include detector normalization background normalization GLP performance check up and actual measurement and evaluation As the counting operations are identical in both modes results obtained in one mode can be used in the other The instrument can for example be normalized in the manual mode and samples evaluated in automatic mode or vice versa Before starting to do normalization read chapter 6 1 Normalization to understand the principle of normalization Select manual mode if it is not already selected Press the START key The display will show Measure manual mode Select the type of measurement or press STOP to cancel measurement Measure assay Normalize background Normalize isotope Do GLP test Press START to measure Select Normalize detectors The list of isotopes is shown see the list in chapter 9 Specifications Select the isotope you want to normalize A display is shown telling you to put the normalization sample in detector number 1 and press the START key If you want to abort this normalization press the STOP key When the display changes to Put the normalization sample in detector number 2 and press the START key Put the source to the indicated detector and press the START key Continue this way until the source has been in each detector Do the normalization for
7. totals references zero sample standard samples unknowns Unknowns are either patient samples or control samples in arbitrary order the controls must be further specified on the line called CONTROLS 2BLANK 2TOTAL 2STD 1 2STD1 2 5 2STD1 5 2STD1 20 2UNKN The number on the left is the replicate number and the number on the right is the nominal concentration The replicate number preceding UNKN means that all unknowns have this number of replicates In the example above all samples are duplicates and no REFER samples are used 67 5 Operation with external MultiCalc Other output definition available with softkeys F2 to F7 and F8 F5 and F8 F7 are COMMENT FACTORS BEGIN INPUT OUTPUT OPTIONS PATIENT and COUNTER For more details about MultiCalc assay protocol parameters see the MultiCalc User Manual Protocol parameter setting A list of available output items for the MultiCalc assay protocol is shown in section 5 8 at the end of this chapter Entry of parameter values in a protocol is supported by various softkeys described in the MultiCalc User manual in the part referred to above 5 4 11 Exiting assay protocol editing When you have finished protocol editing you exit by pressing key F9 and then normally selecting F1 FQUIT SAVE The protocol then joins the list of available MultiCalc protocols To return to the main menu you must press F9 5 4 12 MultiCalc protocol operations If you want to manipulate MultiCalc p
8. Isotopes 1 129 Counting window is Dynamic 166 10 3 RIA IRMA RATIO assay counting The following values are printed in diagnostic info Level is the minimum value that the parameter SYSTEM Diagnostic output Level of diag info must have in order that the value is printed Assay indicates whether a value is printed also during a RIA IRMA RATIO assay or only during isotope normalization Isotope indicates whether a value is printed for both isotopes in a dual labelled assay ECO EEC MINI mensas fr fe Due eee 7 fe roe Prien 6 ve De messes o ve vee wee fo fe D Dead time factor is the factor by which measured counts must be corrected to account for pulses that are lost because they occurred when a previous pulse was being processed You can include the dead time factor in a RIA IRMA RATIO assay printout by setting the parameter SYSTEM Diagnostic output Print dead time factor to Yes Open window counts is the total number of counts in all channels of the multi channel analyser It also includes those pulses that are beyond the MCA high energy limit Spectrum assessment can have one of the following values Spectrum OK Bad spectrum No coincidence peak Too small coincidence peak Too low activity this can appear only during isotope normalization Too high activity Unexpected peak gives the peak channel number of a peak that was not expected In this case Spectrum asse
9. Run id 6 DIAGNOSTIC INFO Dead time factor 1 00011 Open window counts 340 Spectrum assessment Spectrum OK ISOTOPE I 129 Expected peak chn 32 2 Low channel limit 24 High channel limit 41 Used peak channel 31 5 Peak begin channel 1 Peak end channel 42 This peak has noise Window shifted Peaks in smoothed spectrum that exceed 7 Channel Counts 31 18 lt lt END OF DIAGNOSTIC INFO 1 1 20 217 643 0 7 35 2 1 2 1 20 9239 27736 3 1 47 3 1 3 1 20 5586 16800 4 1 76 4 1 4 1 20 3898 11711 4 2 03 5 1 5 1 20 2262 6748 4 2 54 DIAGNOSTIC INFO Dead time factor 1 00090 Open window counts 2822 Spectrum assessment Spectrum OK ISOTOPE I 129 Expected peak chn 31 5 Low channel limit 24 High channel limit 40 Used peak channel 32 2 Peak begin channel 1 Peak end channel 55 This peak has noise Window shifted Peaks in smoothed spectrum that exceed 46 Channel Counts 2 216 lt lt POS RACK DET BATCH TIME COUNTS CPM ERROR 1 1 3 END OF DIAGNOSTIC INFO POS RACK DET BATCH TIME COUNTS CPM ERROR 6 1 1 2 20 2667 8031 6 2 36 7 1 2 2 20 2596 7766 2 2 40 8 1 3 2 20 226 673 2 7 13 9 1 4 2 20 14997 45267 7 1 24 10 1 5 2 20 2332 6958 6 2 51 END OF ASSAY Figure 10 Diagnostic info printout of a CPM assay 165 10 3 RIA IRMA RATIO assay counting NORMALIZATION OF 4 I 129 26 Aug 1994 10 25 45 DIAGNOSTIC INFO FOR DETECTOR 1 ISOTOPE I 129 Actual coverage 63 42 END OF DIAGNOSTIC INFO Total counting time 45 Repeat times De
10. SCV coefficient of variation FLAG concentration flag chn A 49 4 Operation with internal RiaCalc WIZ The items are printed from left to right a single row of results for each sample If replicate samples are employed then the average results are printed on a separate line after individual sample values You can delete an item from the list or add new items to the list 4 6 5 Deleting a printout field Assuming for example that you want to delete the field FLAG from the list Select FLAG and press E A menu will appear on the screen Change item type FLAG Cut this item Paste item after this one Make new item after this one Note the paste item line will only appear if an item has previously been deleted Select Cut this item and press E The program will then show the new list FLAG is not available any longer 4 6 6 Inserting a printout field Lets assume that you want to insert the field ETIME to the previous list between CPM and CONC Select CPM and press EDIT A menu will appear on the screen Change item type CPM Cut this item Paste item after this one Make new item after this one Select Make new item after this one and press E twice A list output items will appear on the screen See the list on the previous page Use the arrow keys to move through the list to the item you want e g in this case ETIME is the sixth item on the list Note You can also use the following keys on t
11. 1 retry 2 omit PRINTER timeout l retry 2 omit 105 7 3 System mode PC not connected l retry 2 omit PC not ready 1 retry 2 omit PC timeout l retry 2 omit Either correct the fault and select 1 retry or disable the expected connection with 2 Fomit Note PC and mainframe port timeouts do not cause output to these ports to be turned off If a timeout occurs a message informing about this stays on the instrument display until you select whether to retry or terminate transmission to this port The instrument waits until the selection is made and no data is lost 7 3 11 Version If you select this you can see what version of the software is being used to control WIZARD 7 3 12 Disk operations 7 3 12 1 Format disk To format a disk for use as a data disk put the disk in the drive and select the Format Disk option If you confirm this operation the disk will be formatted The formatted disk will hold 1 44 Mbytes of data in normal MS DOS format Note If the disk was not an unformatted empty disk then the format procedure will wipe clean all the data on the disk 7 3 12 2 Verify disk This function checks that the disk is properly formatted and readable 106 7 4 STAT counting 7 4 STAT counting STAT counting allows you to interrupt counting of the current batch load new samples manually and count them Results will be briefly displayed on the screen and will also be printed out The STAT samples can then be r
12. PROTOCOL area of the ID clip The rack must have only one holder and sample in the last position of the rack The 1480 counter model can also normalize isotopes at the beginning of a multiple isotope assay MIA This is done if an isotope standard used in MIA has a replicate number greater than 0 A GLP TEST measurement is otherwise similar to isotope normalization only the obtained results are not used in assay counting but are stored separately so that they can be compared with the results obtained in previous GLP TEST measurements GLP means good laboratory practice In the 1470 counter model isotope normalization can also be started manually when the conveyor or rack id readers are not used 10 2 1 What is done during isotope normalization e Peak resolution is determined if SYSTEM Isotope lt isotope name gt Counting window is Dynamic or Dynamic keV It is the half height width of the isotope main peak divided by the energy of the peak e e Detectors are tested for stability if SYSTEM Isotope lt isotope name gt Repeat times is greater than 1 This is not done in 1480 MIA isotope standard normalizations e Background activity in the counting window is updated to correspond to the new counting window e The MCA channel number of the isotope main peak is determined if Counting window is Dynamic or Dynamic keV This makes it possible to determine the effective gain keV channel during normalization T
13. 0 43 2 1 0000 35478 1 0019 0 42 3 1 0000 35377 0 9991 0 43 4 1 0000 35356 0 9984 0 43 5 1 0000 35585 1 0050 0 42 VALUES SAVED END OF ISOTOPE NORMALIZATION Figure 7 1470 normalization printout when SYSTEM Isotope 1 129 Counting window is Dynamic and SYSTEM Printout selections Extended norm printout is No Detector 6 has been set inactive hence only detectors 1 to 5 can be used NORMALIZATION OF 3 Cr 51 22 Sep 1994 17 24 18 Total counting time 30 Repeat times 2 Nominal gain 1 00 keV channel Main peak 320 0 kev Main peak at nominal gain 335 0 channels Nominal window coverage 80 00 PEAK PEAK RESOL WINDOW keV DECAYED MEASURED DETECTOR RELATIVE SIGNIF 000 DET CHN DEV LOW HIGH ACTIVITY COUNTS EFFICIENCY ERROR LEVEL 1 326 3 2 6 8 80 7 347 6 0 9999 19255 0 0000 0 00 0 000 2 319 0 4 8 7 9 82 4 347 8 0 9999 19084 0 0000 0 00 0 000 3 324 5 3 1 8 9 82 0 350 7 0 9999 19040 0 0000 0 00 0 4 336 5 0 5 8 9 82 5 351 0 0 9999 18878 0 0000 0 00 0 000 5 337 0 0 6 8 2 84 0 349 5 0 9999 19003 0 0000 0 00 0 000 6 327 7 2 2 8 9 83 8 348 0 0 9999 18988 0 0000 0 00 0 000 7 326 4 2 6 8 4 80 7 349 6 1 0000 19075 0 0000 0 00 0 000 8 343 5 2 5 8 3 83 3 346 2 1 0000 18846 0 0000 0 00 0 000 9 Failed unexpected peak 10 330 7 1 3 8 1 83 9 346 6 1 0000 19178 0 0000 0 00 0 000 AVG 19039 0 0000 0 00 VALUES NOT SAVED END OF ISOTOPE NORMALIZATION Figure 8 A printout of a Cr 51 normalization that ha
14. 1 in communication protocols WIZARD C00 or WIZARDBG C00 you can use isotope codes 1 99 in single evaluation assay protocols In dual evaluation assays having the assay protocol parameter 01 DUAL ASSAY YES you can only use isotopes 1 9 in this case you concatenate the two isotope codes to form a two digit number Number 3 is reserved to be equal to 12 How to measure the isotope number 3 in single label is explained in the next paragraph If in a dual evaluation assay the second digit of the isotope number is 0 it means that the channel B isotope has code 10 If you set DEFINE WIZARD 0 you can only use isotopes 1 9 in both single and dual evaluation assay protocols However even for single evaluation assays you can specify two isotopes by concatenating the two isotope codes to form a two digit isotope number In this case WIZARD uses two counting windows and the results can be retrieved during assay evaluation in variables COUNT CPM and COUNT B CPM B respectively Single evaluation assays can only use one standard curve even if two isotopes are used As before the number 3 is reserved to be equal to 12 To measure the isotope number 3 in single label use the number 33 and set the assay protocol parameter 01 DUAL ASSAY NO The communication protocol WIZARD_T C00 cannot be used to send measurement parameters to WIZARD so the constant WIZARD does not appear in it If you are using
15. 3 1 2 2 At least one of the isotopes has crosstalk correction 1470 only If at least one of the isotopes has SYSTEM Isotopes isotope name gt Crosstalk correction set to Yes then crosstalk is corrected for in addition to spillover In this case every time a normalization sample is measured in a detector spectra from all detectors used in the counter are stored Possible repeat measurement spectra are summed This makes it possible to determine how much crosstalk there is between detectors for this isotope To do this the program determines counting windows in terms of MCA channels for both isotopes in all detectors used This is done by converting the stored keV values of the counting window boundaries by using the effective gain during the isotope normalization that was used to obtain the spectrum in which counts are summed 159 10 3 RIA IRMA RATIO assay counting Next the activities in both counting windows for all detectors used and for all normalization sample measurements in both normalizations are determined This is done by summing counts over counting windows The results are corrected for dead time converted to CPM the background activity is subtracted and then they are divided by the relative detector efficiency and if selected corrected for isotope decay Dead time background activity and possible isotope decay correction factor are determined by the detector and the isotope spectrum used the relative detector effic
16. 30 841 68 686 846 32 Pb 203 Lead 279 31 52 1 34 Ru 103 Ruthenium 497 30 944 45 400 600 35 Sb 125 Antimony 428 10 2 37E 04 88 37 Sc 47 Scandium 160 80 82 1 75 38 Se 75 Selenium 265 31 2880 93 39 Sm 153 Samarium 103 86 47 40 Sn 113 Tin 392 43 2760 36 350 430 41 Sr 85 Strontium 514 8 1530 50 42 Sr 87m Strontium 388 12 2 8 90 43 Tc 99m Technetium 140 86 6 02 44 Open 15 1000 keV 513 100 0 1024 46 Ge 68 Germanium 512 100 6504 20 1800 47 C 11 Carbon 511 100 3 41E 1 20 1800 48 O 15 Oxygen 511 100 3 40E 2 20 1800 49 N 13 Nitrogen 511 8 1 655E 1 20 1800 50 TI 201 Thallium 70 100 73 06 60 90 51 Cu 64 Copper 511 8 12 701 425 640 52 Ti 45 Titanium 511 8 3 08 420 600 53 Re 188 Rhenium 155 70 16 98 97 95 203 91 I 125T lodine GLP 29 82 1445 25 92 Cs 137T Caesium GLP 662 5 6 2 63E 5 93 Ge 68T Germanium GLP 512 100 6912 20 1800 Eff CPM DPM x 100 typical values open window It includes transition probabilities 128 10 CALCULATION METHODS Applies to program version 3 5 August 1999 129 10 1 Background normalization 10Calculation methods 10 1 Background normalization 10 1 1 How background is measured When an empty rack having an ID clip with the BKG label sticked in the area marked RACK SPECIAL is encountered during automatic measurement counts are collected from all detectors for the time given by the parameter SYSTEM Background Bgrd counting time The obtained spectra are then
17. 60 3 382 0 24 7 51 5 188 3 7 8 044 fo Roker ol 71 14 34 49 5 5 7 9 0 412 5 34 7 58 2 194 0 14 7 0 5 5 8 0 O0 OMOM VALUES SAVED END OF BACKGROUND NORMALIZATION Figure 1 Background normalization printout when SYSTEM Diagnostic output Print diagnostic info is Yes and only detector 2 has been selected in the menu SYSTEM Diagnostic output Select detectors for info and SYSTEM Printout selections Horizontal bgrd printout is No BACKGROUND PRINTOUT Measured on Counting time Counts per minute DET OPEN 409 413 382 374 382 388 412 411 395 368 WODAIHDUEPWNHE OOoOoooooooo eo NOEL QS 10 37 06 END OF BACKGROUND PRINTOUT OY iS UT UID AS 14S O 306 010 01 OY Ul 000 009 EO 0 OO NBROOCOOWOOrF m o N w gt H m w wo H Figure 2 The same background normalization is printed again using SYSTEM Backgroud Print background Now SYSTEM Printout selections Horizontal bgrd printout is Yes The width of this layout will not exceed 80 characters if there are at most 10 isotopes that are normalized 134 10 2 Isotope normalization and GLP TEST measurement 10 21sotope normalization and GLP TEST measurement When a rack having an ID clip with the NORM label stuck in the area marked RACK SPECIAL is encountered during automatic measurement WIZARD starts normalising the isotope that has the code number that is in the
18. AXIS response 22 FITTING ALGORITHM 23 STD OUTLIER REJECTION 24 HALT FOR CURVE EDIT 66 5 Operation with external MultiCalc These are all to do with the standard curve used for determining concentration values The built in MultiCalc helps explain these parameter along with the part in the MultiCalc User Manual called Standard curves 5 4 8 Quality control parameters The next block of parameters are concerned with quality control They are 60 CONTROLS 61 HISTOGRAM 62 OC ACCEPTANCE RULES See the part on Quality Control in the MultiCalc manual 5 4 9 Output parameters The final block of parameters is concerned with output from MultiCalc 80 STORED FILES These are the types of files stored e g data for further analysis with MultiCalc or other programs or computers curves QC information etc 81 DISPLAY These are the items displayed 82 PRINTER These are the items printed 83 OUTPUT Here you specify what the actual items are in for storing in ASCII format 84 RESULTS Here you specify what the actual items are in the files for external programs or computers 5 4 10 Additional output information The program must know in which order RIA standards are loaded and what their nominal concentrations are This information is supplied in the CODING accessed by pressing F1 Other special samples such as Blank and Reference can also be specified The recommended order for sample tubes is blanks NSB
19. Bi directional automatic sample changer with a storage capacity of 55 racks 550 samples or 100 racks 1000 samples 9 4 4 Sample ID Each rack can be provided with two barcodes each having capacity for 2 digits or a special code word 125 9 Specifications 9 5 Detector system Detectors are of the end well type with a 50 mm x 32 mm 2 0 x 1 1 4 thallium activated sodium iodide crystal 9 5 1 Shielding The detector assembly is surrounded by a minimum of 12 mm of lead shielding against radiation in the vertical plane The shielding against radiation from samples on the conveyor is 30 mm 1 1 4 The shielding between detectors is 7 mm 9 5 2 Detector Matching Within 1 of mean counts of all detectors after normalization 9 5 3 Isotopes The instrument is preset for the isotopes listed in the table at the end of this chapter All isotope settings can be changed by the user 9 5 4 Efficiency 1 125 gt 78 Co 57 gt 80 Cr 51 gt 3 Cs 137 26 typical Co 58 3 5 typical 9 5 5 Energy resolution 1 125 lt 30 Co 57 lt 16 Cr 51 14 Cs 137 10 96 typical Co 58 8 typical 9 5 6 Spilldown Spilldown of Co 57 into I 125 preset regions 3 uncorrected 196 corrected 9 5 7 Crosstalk Crosstalk detector to detector uncorrected worst case 1 125 negligible Co 57 negligible Cr 51 lt 0 5 Cs 137 lt 4 96 Co 58 5 96 Crosstalk conveyor to detector si
20. Counting window is Fixed a spectrum is never considered to be bad 10 3 5 TOTAL CPS and TOTAL DPS These values are stored in the file WASTELOG TXT in the root directory of the instrument hard disk TOTAL CPS is the sum of all printed corrected CPM values of measured tubes in the assay The DPS value is obtained by dividing the printed corrected CPM value by the actual coverage of the isotope counting window and by SYSTEM Isotope lt isotope name gt Efficiency TOTAL DPS is the sum of all DPS values of measured tubes in the assay 10 3 6 Storing the latest counting window At the end of the assay the latest observed isotope main peak position in the MCA channels is stored for each detector used and isotope if the assay is dual labelled This is used as the expected peak position in the next assay using the same isotope until a sufficiently strong isotope peak is observed again This is done even if the parameter SYSTEM Operating mode Default is norm window is Yes 10 3 7 How the relative CPM error is calculated in RIA IRMA RATIO CPM printout In CPM printout this quantity has the title ERROR The following formulas are used Variance1 SQUARE DeadTimeFactor 60 CountingTimelnSeconds CountsinWindow BVariance SQUARE BgrdDeadTimeFactor 60 BgrdCountingTimelnSeconds BackgroundCountsInWindow Variance2 Variance1 BVariance RelativeError1 SQUAREROOT Variance2 CorrectedCpm1 Rela
21. E Edit protocol 05 BF_BLAN 06 FOLATE 07 FORMAT 08 LEARN 09 PROTOI 10 PROTO2 11 PROTO3 12 READ ME 13 PROTOO 14 TEST_1 15 TEST_3 16 TSH Id order change with lt gt You may now change the parameters You can move up and down in the parameter list by means of the UP and DOWN arrow keys See section 3 5 for details of parameters available in a protocol 30 CPM operation Note In the example above the protocol types include both RIA and RATIO It is not possible to create RIA protocols in CPM mode protocols are automatically labelled RATIO but if such a protocol has been created in another mode then it will still appear on the list You can select any protocol from the list for editing in CPM mode but only the three parameters shown in the example below will appear Edit protocol 11 PROTO3 Max counts limit Labels Copy An existing protocol can be copied so as to create another protocol with the same contents Select Copy and press E Select the protocol to be copied Give the name the new protocol is to have You may also give an ID number Select Do copy and press E Rename This option allows you to give a new name to a protocol Select Rename and press E Select the protocol to be renamed from the list of protocols Give it a new name and or ID number Select Do rename and press E Delete Select Delete and press E Select the first protocol to be deleted Then select Do delete
22. Isotopes lt Isotope name gt Normalization time and the printout is similar to the isotope normalization printout If SYSTEM Isotopes isotope name gt Repeat times is greater than 1 the sample is measured in each detector as many times as specified in Repeat times The total counting time for the sample in each detector is in this case Repeat times multiplied by SYSTEM Isotopes isotope name gt Normalization time The following values are saved during GLP TEST normalization e Isotope main peak channel number e Background CPM in the counting window e Relative detector efficiency if a 1470 counter has more than one detector installed e Detector resolution e Absolute detector efficiency This is determined for I 125 Dynamic and Dynamic keV windows using the Horrocks method For other isotopes and for I 125 Fixed window the efficiency is calculated by dividing the measured corrected CPM in the counting window by the absolute activity of the test sample which is given by the parameter SYSTEM Isotope lt isotope name gt GLP test sample DPM An isotope is considered to be I 125 if SYSTEM Isotope lt isotope name gt Spectrum type is I 125 See Section 10 2 4 2 4 on page 142 The corrected CPM above is corrected for dead time background activity and isotope decay but not for relative detector efficiency 149 10 2 Isotope normalization and GLP TEST measurement e Window coverage T
23. LAG 0 111 Concentration flag all values but only unknowns LAG B 0 DIT As above for channel B K 111 BLANK cpm valid after BLANK in coding all values for both groups KB 111 above for channel B ER 111 above for REFER ER B 111 above for channel B OTAL 111 previously for TOTAL OTAL B 111 above for channel B EG 111 above for negative controls EG B 111 above for channel B TRSTHST S 111 above for positive controls c r oo nn B 111 above for channel B ESP 11 Programmable ESP B 11 As above for L B 0 111 Numerical flag number These are 1 OUT 2 gt STD 3 lt STD 4 amb 5 gt gt STD 6 lt lt STD 7 CV 8 CV 9 gt gt 3CV STS B 44 0 111 Numerical flag number channel B GROUP GR 0 11 Group number of multiple UNKN coding REPL RP 0 1 Replicate of individual sample for unknowns SAMPLE SPL 0 111 Sample No includes controls for all values but only unknowns RESP 11 Response error RESP B 11 Response error for channel B ILF 0 11 Dilution factor EMARK diia HEAR 0 111 Remark EOA 1 Application sequence OW 111 Row number 1 individual 2 average 3 second average ATE 111 Date LOCK 111 Time LASS 444 4 111 E g 1 if CONC STD 1 or 2 5 if CONC STD 3 STD 2 2 LASS B 111 As above for channel B I
24. Note if you load a new protocol when a previous one with the same name or ID number exists in the instrument you have the choice of either renaming the protocol to be loaded or deleting the previous protocol Save This function allows you to transfer files from WIZARD to an external microfloppy disk for storage Subsequently these files can either be loaded back into WIZARD Purge Deleted protocols and associated data files can be permanently erased from the instrument hard disk with this function Password When a protocol is created you can give a two character password All characters are allowed in the password and the password is case sensitive If you do not give a password at this point then the protocol can be edited renamed overwritten when a new protocol with the same name or ID number is loaded and deleted without giving any password Otherwise the password is needed to do these operations A protocol can always be copied restored saved and purged without giving the password With rename it 1s possible to change or remove the password 32 CPM operation Passwords are retained even if power is turned off The password feature can be disabled by removing the file C PASSWORD from the instrument hard disk To enable it again type ECHO aa gt CAPASSWORD at the DOS prompt to recreate a password file and boot the instrument You can enter DOS by installing the installation disk to the disk drive and by restarting the instru
25. SYSTEM Isotope lt isotope name gt Peak pos keV and SYSTEM Isotope lt isotope name gt Window coverage For window types Dynamic keV and Fixed the boundaries are given by the isotope parameters SYSTEM Isotopes lt isotope name gt Low boundary ke V and High boundary keV If Counting window is Dynamic or Dynamic keV then the effective gain during isotope normalization is calculated by dividing the energy of the main isotope peak in keV s by the MCA channel number of the main peak If Counting window is Fixed then the nominal gain is used as the effective gain Next the activities of both isotopes in both counting windows are determined in both normalizations The following is repeated for each detector that is in use For both isotope spectra counts are summed over both isotope counting windows Sums are corrected for dead time converted to CPM the background activity is subtracted and the result is divided by the relative detector efficiency and if selected corrected for isotope decay 158 10 3 RIA IRMA RATIO assay counting Dead time background activity and possible isotope decay correction factor are determined by the detector and the isotope spectrum used the relative detector efficiency by the detector and the isotope counting window used Detector efficiency depends on the counting window used and may also change with time We assume that the energy dependency of a detector s efficien
26. To set only the date send the command CLOCK dd mo yy To set only the time send the comman CLOCK hh mi ss or CLOCK hh mi The date is not accepted in any other formats than dd mo yy The year must be a two digit number The other values must also be expressed as two digit numbers e g the month of May is 05 Values from 80 to 99 refer to the 20th century and values below 80 to the 21st century The time must be in 24 hour format After the date has been set the counter responds with the string DATE SET After the time has been set the counter responds with the string TIME SET If for some reason the date and or time string could not be interpreted the counter responds with 7 Bad date or time string If date or time is set as both normally are the sending of time of day values with RIA IRMA RATIO assay results to MultiCalc is also enabled The exact time when measurement of each sample was started can also be sent to MultiCalc In MultiCalc mode the measurement start field can be enabled and disabled only from MultiCalc the SYSTEM parameter Diagnostic output Print meas start time has no effect in this case To enable the sending of measurement starting time with assay results send from MultiCalc terminal the command CLOCK ON to the counter To disable it send the command CLOCK OFF The sending of this command can be made automatic if it is included in the WIZARD communication protocol by setting the parameter
27. USECLK to 1 See the information that appears when you select Fl COUNTER F5 INSTALL The clock value will be assigned to the parameter COUNT_B This is only available for this use in single label counting You can check the current setting by sending the command CLOCK If the measurement starting time has been enabled the counter responds with the string PRINTED 71 5 Operation with external MultiCalc If it has been disabled the counter responds with the message NOT PRINTED When the counter is turned on or after a power failure in MultiCalc mode the measurement start time field is disabled until it is explicitly enabled again from MultiCalc 12 5 Operation with external MultiCalc 5 8 Selectable outputs RACK 1 Rack number 3 digits for both groups no average DET 1 Detector number for both groups no average SEO 111 Sequence or tube number PAT 0 111 Patient number with both averages unknowns only TIME 1 Counting time in seconds ETIME 1 Elapsed time in decimal hours COUNT 11 Total counts and first average COUNT B 11 Total counts and first average on B channel Htttit 11 Corrected CPM and first average B 11 Corrected CPM and first average on B channel Hitt HHF 111 Concentration with both averages B 111 As above for channel B O11 Coefficient of variation as a percentage only averages B O11 As above for channel B E
28. Unkn multipl factor Unkn outlier lituana da V Verify disk ccoo iris iii aan 106 Version of software sss 106 Vals C A 120 Din SAO sii la GS 125 Voltage selection iiie ets eres etae eoo innen 184 W Warning assay deviation 96 sess 96 Waste log file sss 35 58 70 Window coverage 5 135 141 ho A neue 120 Without buffering to PC 69 99 Write results to file oooooconococonnnonnnononcconcconoconnnonnnoon 99 X B CP T CREER RR EAA EEE 43 Y BE S REP TE MEET 43 199 13 Index
29. WIZARD A disk can be used for transferring results from WIZARD RiaCalc WIZ operation of WIZARD R Fix ID clip here Position 1 1 Fix ID clips to racks E ID labels barcodes are stuck to an ID clip which fits onto a rack to tell WIZARD the function of the rack A counting protocol is a set of three parameters time max counts limit and isotope which control counting Rack number is optional and allows each rack to have its own Irzexzsrecud snorocor P Dor rotocol number or isotope Normalization ensures that the counting efficiency of each manet detector is the same 07 18 Rack number or NORM Background ensures that the effect of the background is removed TONNE OL TEST BKG or STOP i from the measured counts wistuetion Test initiates a GLP performance test normalization Isotope number shows the isotope to be used in normalization Stop tells WIZARD that no more racks are to be counted au ID clip faces forward when Position 1 loading racks Make sure racks are loaded the correct way round with the ID clips facing away from you as shown in the figure Start by loading the right hand side of the conveyor 3 Press START to count Dee Live display during counting is obtained by selecting from the main menu on the WIZARD built in display Operate and then Show cpm results Available live displays are counting para meters Counts CPM CPS and Spectrum RiaCalc WIZ output can also
30. a counting window counts from the low and high boundary channels are added only in the proportion that these channels are included inside the counting window 10 1 6 Diagnostic info Diagnostic info is printed during background normalization if SYSTEM Diagnostic output Print diagnostic info is Yes For 1470 you can select the detectors for which diagnostic info is printed in the menu SYSTEM Diagnostic output Select detectors for info The following values are printed For all detectors that are in use e Open window counts e Dead time factor For all normalized isotopes and for all detectors that are in use e Effective gain keV channel e Counting window in MCA channels e Measured counts in the counting window 133 10 1 Background normalization BACKGROUND NORMALIZATION 22 Sep 1994 10 37 05 DIAGNOSTIC INFO DETECTOR 2 Open window counts Dead time factor ISOTOPE 1 I 125 Iodine DETECTOR 2 Gain Window channels Counts in window ISOTOPE 2 Co 57 Cobalt DETECTOR 2 Gain Counts in window ISOTOPE 3 Cr 51 Chromium DETECTOR 2 Gain Window channels Counts in window ISOTOPE 4 I 129 Iodine DETECTOR 2 Gain Window channels Counts in window END OF DIAGNOSTIC INFO Counting time Counts per minute DET 1 2 OPEN 409 0 413 0 I 125 38 0 32 4 Co 57 56 5 49 6 Cr 51 190 8 199 8 I 129 24 1 7 1 ES channels 413 1 000 0 957 17 5 32 0 941 94 7 49 1 004 83 1 201 0 946 19 6 7
31. and press E The protocol and any associated data will then be deleted Recover Deleted protocols can in most cases be recovered by selecting this option They are saved in the protocol index area There is room on the system disk for 100 deleted and active protocols The larger the number of active protocols the fewer the number of deleted protocols that can be saved If there is no room the oldest deleted protocol will be permanently removed to make room for a newly created one No protocol with the same name should have been deleted later than the one you want to recover 31 CPM operation Print With this function you can print the contents of a single protocol or the protocol index according to the selection you make Isotope names are included in the protocol printout For dual label protocols the code numbers for both isotopes are printed separately e g 1 I 125 2 Co 57 Load You can load a single protocol or all protocols from an external microfloppy disk Such protocols will have been saved there using the Save function see below Note if you load a new protocol when a previous one with the same name or ID number exists in the instrument you have the choice of either renaming the protocol to be loaded or deleting the previous protocol Normal Load will give you Protocol files 2 Controls files 3 Trends files Extended Load will in addition give you 4 Standard curve files 5 Data files 6 Results files
32. are in grey they are only available in RiaCalc WIZ however the following three are 5 6 1 Spectra Store spectra this is for information only It tells if spectra can be saved or not and if they are to be sent to an output device or not The actual settings can be made in SYSTEM Operation mode Store assay spectra See there for more details Operation this allows you to handle the files of spectrum data The options are Delete delete the spectrum file data Save to disk save the spectrum information on the program disk in WIZARD Send to PC send the spectrum information to a PC that is connected to WIZARD Nr of channels the number of channels that are included in the spectra During assay measurement all spectra are stored using the full 1024 channels This file is in binary format If you are only interested in the lower part of spectra you can set here the number of channels that are included when the file is converted to text form The channels included always start from the first channel Format When assay spectra are saved in the datalogger disk or are sent to PC or mainframe you can specify with this parameter the format of the data Available formats are Wallac Excel or Ortec Wallac format This is a text file that can be read by Wallac Spectrum Analysis Program Excel format This text file format data belonging to the same spectrum is written in the same line and channel counts values are
33. be evaluated again In addition to editing the data before evaluation you can change the curve fitting method during evaluation if you have selected Halt for curve edit in protocol setting 4 9 4 Results files Results files comprise data but they are in ASCII format and are intended for output to an external PC or mainframe which is able to handle the widely accepted ASCH format View Lets you view but not edit a file Print Lets you print a file Send to PC Lets you send a file to PC Delete Lets you delete a file Save to disk Lets you save result files in the EVAL directory of a micro floppy disk When you have a View a result file you can move up or down one page by pressing the PgUp or PgDn keys respectively and to the beginning or end of the file by pressing the Home or End keys respectively 55 4 Operation with internal RiaCalc WIZ Note the view mode only shows the first 40 characters of any line in the Result file and does not allow any editing of the values 4 9 5 Standard curve files Standard curves are produced in RIA and IRMA and can be viewed with this function Note For standard curve files the run id designation REF means the reference curve View Lets you view a file Delete Lets you delete a file Set reference curve Lets you set a copy of the selected standard curve as the reference curve for this protocol Standard curves can be viewed and if required edited First you must select the curve you wa
34. boundaries are converted from keV to MCA channels and counts are summed over the window Counting window boundaries are determined to a fraction of channel When counts are summed over a counting window counts from the low and high boundary channels are added only in the proportion that these channels are included inside the counting window The sum is corrected for dead time converted to CPM the background activity is subtracted and this value is divided by the relative detector efficiency then correction is made for isotope decay if such has been selected Assay measurements are corrected for isotope decay if SYSTEM Isotopes isotope name gt Decay correction is Yes and SYSTEM Isotopes isotope name gt Assay zero time is not None If Assay zero time is Start decay correction is done to the start of the assay The starting time is printed in the title line of the assay printout If the value of Assay zero time is an explicit time decay correction is done with reference to that time After this spillover and crosstalk are eliminated as follows e For single label assays that do not correct for crosstalk the CPM values are printed as such e For dual label assays that do not correct for crosstalk that is for both isotopes SYSTEM Isotopes lt isotope name gt Crosstalk correction is No the CPM values of both counting windows are used to form for each detector a 1x2 vector A as described in Section 10 3 1
35. can abort the power failure recovery operation by pressing the STOP key A menu appears allowing you three choices 1 Continue the STOP instruction is cancelled 2 Stop conveyor the recovery operation is stopped and the instrument returns to the main menu 3 Clear conveyor the counting and transfer lanes of the conveyor are cleared of racks the recovery operation is totally cancelled and the instrument returns to the main menu 7 5 3 Power failure recovery While recovery is occurring a text is displayed telling that power failure has occurred and that operation will begin again from the start of the assay To remove this message press any key The program determines how many assays were uncompleted when the power failure occurred Uncompleted means that although counting of an assay has been started and may even have been ended the results have not yet been either completely saved on disk or sent out to a printer or external computer which of these is valid depends on the options selected This information is stored in a special battery powered memory The conveyor will be then be run forward till the protocol ID cassette of the earliest of these uncompleted assays can be read again The ID is read and counting starts in the normal manner for this assay followed by all the other uncompleted assays in turn The figure see next page shows how the power failure routine works There are two assays on the conveyor the one dark co
36. determined To do the converting the program has to have a value for the effective gain keV channel during this assay The conversion formula is explained in Section 10 2 4 1 2 on page 137 10 3 2 1 Fixed window If SYSTEM Isotope lt isotope name gt Counting window is Fixed the ke V values of window boundaries are given by the parameters SYSTEM Isotope lt isotope name gt Low boundary keV and High boundary keV The nominal gain is used as the effective gain value when these are converted to MCA channel numbers Thus any drifting due e g to temperature variations is not taken into account In dual label counting both isotope counting windows are taken to be fixed if at least one of the isotopes has Counting window equal to Fixed In 161 10 3 RIA IRMA RATIO assay counting this case the nominal gain is used as the effective gain also for the isotope for which the counting window is not fixed Nominal gain is explained in Section 10 1 4 on page 132 10 3 2 2 Dynamic and Dynamic keV window If Counting window is Dynamic keV the ke V values of window boundaries are given by the parameters Low boundary keV and High boundary keV If Counting window is Dynamic the window boundary keV values are determined during isotope normalization on the basis of the parameters SYSTEM Isotope isotope name gt Peak pos keV and SYSTEM Isotope lt isotope name gt Window coverage These w
37. do this 1t needs to be able to identify each batch of samples The WIZARD ID system uses a plastic clip onto which one or two barcode labels are stuck The clip is then clipped onto the rack to be identified The clip can easily be removed and a new one fitted when required The barcodes give information to WIZARD about e g the counting protocol to be used for the samples what the rack number is etc Only the first rack in an assay needs to have a protocol label Barcodes are to be found in a booklet supplied with the counter There are two classes of barcodes numerical ones 0 99 and instruction ones The figure below shows an empty clip and then several examples of clips with labels on them Some have two labels and some only one In addition there is a third area of the clip which comes on the end of the rack This can be used for you to write your own information on The ID label booklet includes empty labels for you to write on and stick to the clip There are also barcodes marked PCURVE and CONTROL which are not used in this program Racks speciat PROTOCOL Empty clip Clip with rack number 02 and protocol number 18 Clip with normalization instruction and isotope code number 02 BKC Protocor PN Ip indicating a background measurement to be made gt JOP PROTOCOL LLLI Clip indicating that counting is to stop after counting this rack if it contains samples If it is empty counting will stop
38. e Relative detector efficiencies if a 1470 counter has more than one detector installed e Crosstalk correction coefficients that are used in single label RIA IRMA RATIO assay counting if a 1470 counter has more than one detector installed Spill and crosstalk correction coefficients that are used in dual label RIA IRMA RATIO assays are calculated at the beginning of the assay so are MIA spillover correction coefficients e Theisotope standard sample activity and standard replicate tube activities if isotope normalization is done at the beginning of a 1480 multiple isotope assay MIA e To test detectors for stability if SYSTEM Isotope isotope name gt Repeat times is greater than 1 10 2 4 1 Fixed window When the boundaries of a fixed window are converted from keV to MCA channels the nominal gain keV channels is used as the effective gain Thus any drifting due e g to temperature variations is not taken into account Nominal gain is explained in Section 10 1 4 on page 132 10 2 4 1 1 When fixed window should be used Fixed counting window is handy in that there is no need to specify the isotope main peak You can use it if you do not know what the isotope spectrum looks like or are otherwise unsure how various isotope parameters should be set for the isotope Using a fixed window you can even measure samples that contain unknown isotopes If you set SYSTEM Isotopes lt isotope name Low boundary keV to 0 and High
39. gives isotope decay corrected counts in repeat measurement CTIME the counting time used in repeat measurement i Actually the counting time is the same for all normalization repeat measurements and MIA standard replicate measurements Note that if SYSTEM Isotopes lt isotope name gt Norm zero time is Start the decay correction is done to the start of the normalization even if the normalization is done at the beginning of a 1480 multiple isotope assay MIA If such an assay has several isotope normalizations all with Norm zero time equal to Start the decay corrections are made to the start of each individual normalization not to the start of the MIA 10 2 9 Calculating results for individual standard replicate tubes in 1480 MIA isotope normalization If an isotope normalization is done at the beginning of a 1480 multiple isotope assay MIA the standard may consist of several replicate tubes The measured replicate tube spectra are summed and this sum spectrum is used to calculate normalization results However the individual replicate tube spectra are used to print a summary info about each replicate tube at the beginning of the isotope normalization printout The following data are printed for each replicate tube e replicate number e counting time for that replicate it is the same for all replicates e measured counts e corrected CPM e ratio of corrected CPM of the individual replicate tube to average CPM of all
40. immediately 22 2 Wizard controls 2 6 Fitting ID labels The figure below shows a rack with an empty ID clip fitted on it There is a special recess on one side of each rack This is where the clip fits It will not fit to the other end of the rack This is important because it defines which way round the rack should be on the conveyor Make sure the ID clip is fitted properly so that it does not slip off when the rack is on the conveyor Empty clip fitted onto rack 2 7 Loading racks the right way round When you load racks on the conveyor the ID labels must face away from you with the white dot on the right side as shown in the next figure Direction of movement on the conveyor White dot 7 Rack orientation when loaded onto the conveyor Information written by the user on the ID label The handwritten information on the label at the end of the rack is then clearly visible from the side of the conveyor All racks even those without labels must be put the same way round with the recess for the label and white spot on the right hand side when you load the racks If it is not the right way round a warning is displayed and counting is stopped until the rack has been turned the right way round 23 2 Wizard controls 2 8 Instruction labels Use instruction labels to select special functions attach the label in the area marked RACK SPECIAL The instrument recognizes the following codes 2 8 1 STOP This st
41. in the command WIA and press Enter After a short while the PC display will show the MultiCalc main menu Please press any softkey or letter from the list E PROTOCOL EDIT F PROTOCOL CREATE DEFAULT PROTOCOL EDIT HISTORY FILE INPUT FILE EDIT INPUT FILE CREATE DELETE ONE RUN PLATE MAP EDIT QUALITY CONTROL REPORT SYSTEM PARAMETERS DISK USE TABLE USER AREA V DISPOSE EXTRA FILES X EXIT TO MS DOS At the bottom of the PC screen you will see eight softkeys labelled F1 to F8 corresponding to the function keys on your computer keyboard also labelled F1 to F8 A softkey changes its function according to the step in the program you have reached The actual function of each key at any time is shown on the bottom of the PC screen The eight softkeys which have functions in the main menu are 63 5 Operation with external MultiCalc F1 COUNTER F2 EVALUATE F3 WORKLISTS F4 PROTOCOLS F5 INP FILES F6 RESULTS F7 LEVELS F8 ETC In addition to the eight softkeys the keys F9 and F10 have fixed functions Function key F9 is always EXIT and F10 is HELP As an alternative to using softkeys there are quick commands listed on the MultiCalc main menu By pressing the appropriate letter you can make the program go directly to a particular function instead of by pressing one or more softkeys MultiCalc supports different levels of operation e g learning or advanced with automatic or optional hel
42. instruction you are telling which counter you are actually going to work with If MultiCalc cannot get WIZARD into the Ready state it will ask you to press F3 to get into the terminal mode Press F3 and try to get WIZARD to the Ready state You can also try pressing the Esc key several times At the end of installation you will see instructions about MultiCalc WIZARD use For more details about MultiCalc installation see the MultiCalc Supervisor s Manual 1224 930 11 9 Functional check Carry out the functional check and the performance test of the instrument For checking the instrument prepare three racks with the following labels BKG NORM 4 STOP Place the 1 129 standard supplied with the instrument in the last position of the normalization rack Remove the holders from positions 1 to 9 in the rack The background rack must be an empty rack with the BKG ID label The STOP rack is a totally empty rack or a rack with a STOP ID label In the latter case it does not matter if there are holders or not Load the racks onto the conveyor in the order BKG NORM and STOP To obtain a full printout follow the procedure below Go to the SYSTEM menu select Printout selections and change Extended norm printout to Yes Then press EXIT and SAVE changes Set SYSTEM parameters Isotopes I 129 Norm counting time s to 180 and Background Bgrd Counting time to 600s Press EXIT and SAVE changes Start counting and check th
43. mode 7 3 3 7 MCA high limit This is given for information and cannot be changed Its value depends on the value of SYSTEM Active hardware Nominal gain 7 3 3 8 Counting window Various parameters affecting the counting window can be set These are Dynamic The boundaries of the window are determined by the parameter Window coverage the window follows peak drifting during normalization and also during assay measurement if SYSTEM Operation mode No dynamic normalization is No Dynamic keV The boundaries of the window are determined by the user in keV the window follows peak drifting during normalization and also during assay measurement if SYSTEM Operation mode No dynamic normalization is No Fixed The boundaries of the window are determined by the user in keV the window does not follow peak drifting Peak position keV This refers to the main photopeak energy Note If the parameter Counting window selected for an isotope in the isotope editor is Dynamic or Dynamic keV then in RiaCalc WIZ assays the window that has been determined during isotope normalization is shifted in accordance with deviation in peak position during normalization and also during assay measurement if SYSTEM Operation mode No dynamic normalization is No If a sample is so weak that its peak position cannot be determined the same counting window as in the previous batch is used If a
44. name or counting time A MultiCalc protocol is used mainly to define the way the data from the counter is to be handled e g items to be output quality control curves to be plotted etc These parameters are described in detail in the MultiCalc User manuals The line 3 parameter is called Measuring parameters This is where you give the isotope number See 5 4 3 Isotope selection and 9 8 Isotopes defined for 1470 WIZARD You just select the number of a suitable isotope from the list of those available Make sure that the one you select has been normalized If it has not you need to make one as described in Part 6 Normalization 5 4 Editing a MultiCalc protocol Starting from the MultiCalc main menu and depending on whether you want to edit an existing protocol or create a new one you can either press the letters E for protocol edit or F for protocol create or you can press softkey F4 PROTOCOLS followed by F1 EDIT or F2 CREATE You must then select a protocol number from the list of protocols In the case of EDIT you must select an existing protocol but in the case of CREATE you select must glve a new protocol and number Usually this number will be the next one in the number sequence but you can glve a different number if you want You must then select the Technology which in this case is Gamma If you create a protocol you must then specify the protocol type RIA IRMA or RATIO This choice determines the type of parameter list you get Th
45. number Normalization ensures that the counting efficiency of each detector is the same m Rack number or NORM Background ensures that the effect of the background is TEST BKG or STOP removed from the measured counts instruction Test initiates a GLP performance test normalization Isotope number shows the isotope to be used in normalization Position 1 Stop tells WIZARD that no more racks are to be counted 2 Load racks onto WIZARD Make sure racks are loaded the correct way round wnth the ID clips facing away from you as shown in the figure Start by loading the right hand side of the conveyor 3 Press START to count Live display during counting 1s obtained by selecting from the main menu on the WIZARD built in display Operate and then Show cpm results Available live displays are counting parameters Counts CPM CPS and Spectrum 4 Results are printed out The built in program allows counting and normalization protocols to be created and edited The built in display and keyboard is used Results are sent to the display and a printer Racks speciar PROTOCOL Protocol number or isotope number ID clip faces forward when loading racks Live display available during counting For System setting info e g detector deactivation or clock setting select SYSTEM in the main menu The printer connected to WIZARD port 1 is used for printing corrected CPM results directly from
46. on page 137 may not be entirely accurate the implementation chosen for MIA in practice gives better results The following NxN matrix of CPM activities is set up where N is the number of isotope standard samples 171 Spectrum of isotope standard 1 Counting window of isotope standard 1 Spectrum of isotope standard 1 Counting window of isotope standard 2 Spectrum of isotope standard 1 Counting window of isotope standard N 10 4 Multiple isotope assays MIA Spectrum of isotope standard 2 Counting window of isotope standard 1 Spectrum of isotope standard 2 Counting window of isotope standard 2 Spectrum of isotope standard 2 Counting window of isotope standard N Spectrum of isotope standard N Counting window of isotope standard 1 Spectrum of isotope standard N Counting window of isotope standard 2 Spectrum of isotope standard N Counting window of isotope standard N Then values in each column are divided by the diagonal value in that column This converts measured activities to relative spillover values Let us denote this new matrix with the letter M If this matrix is multiplied with the Isotope 1 activity Isotope 2 activity Isotope N activity following 1xN vector B the result is the following 1xN vector A Measured activity in the counting window of isotope 1 Measured activity in the counting window of isotope 2 Measured activity in the counting window of isotope N Thu
47. p2 90000000000 0000000000 110 7 6 Routine maintenance 7 6 Routine maintenance 7 6 1 Cleaning The instrument and its environment should be cleaned regularly to avoid contamination and raised background In particular the conveyor and elevator forks should be cleaned using a soft cloth and alcohol Note The dust covers should be kept closed to reduce the amount of dust entering the detector wells 7 6 2 Checking the background If you want to check the background without actually saving the results as will happen in background normalization then you must count an empty rack which has holders but no sample tubes Make sure the counting time is long enough to collect enough counts e g 10 min 7 6 3 Decontamination If the background does increase then you should carefully clean the detector wells using a soft cloth soaked in alcohol Avoid using any decontamination liquid which might be corrosive Beware of scratching the surface of the detector wells Check that the background is back to normal after cleaning It can also be that the sample carriers become contaminated These should be regularly checked and if necessary cleaned or replaced A mild decontamination solution can be used for this 111 7 7 Safety information 7 7 Safety 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 compreh
48. password is case sensitive If you do not give a password at this point then the protocol can be edited renamed overwritten when a new protocol with the same name or ID number is loaded and deleted without giving any password Otherwise the password is needed to do these operations A protocol can always be copied restored saved and purged without giving the password With rename it is possible to change or remove the password Passwords are retained even if power is turned off The password feature can be disabled by removing the file C PASSWORD from the instrument hard disk To enable it again type ECHO aa gt CAPASSWORD at the DOS 42 4 Operation with internal RiaCalc WIZ prompt to recreate a password file and boot the instrument You can enter DOS by installing the installation disk to the disk drive and by restarting the instrument 4 5 Parameters available 4 5 1 Dual evaluation If you do normal single isotope RIA IRMA select NO If you would like to measure samples labelled with two isotopes in the same vial select YES See Dual label counting for more details 4 5 2 Counting time Give the counting time in seconds All samples are counted for this time 4 5 3 Max counts limit If you want to terminate counting on the basis of the number of counts accumulated enter the counts value for this parameter Provided this number of counts is reached in each detector before the counting time expires this parameter will terminat
49. possible to do GLP test measurements by only using the isotope numbers 4 91 92 and 93 If needed it is also possible to make other isotopes available for GLP test measurements 6 2 3 Saved GLP values The following values are saved during GLP test normalization PEAK Isotope main peak channel number BGRD Background CPM in counting window EFFIC Relative detector efficiency ifa 1470 counter has more than one detector installed RESOL Detector resolution EFFICIENCY Absolute detector efficiency This is determined for I 125 using the Horrock s method For other isotopes the measured CPM in the counting window is divided by the absolute activity of the test sample which is given by the SYSTEM parameter Isotope lt Isotope name gt GLP test sample DPM COVERAGE Window coverage This is the fraction of counts in the whole spectrum that falls to the isotope counting window CHI PROB Detector stability probability This can be calculated ifthe SYSTEM parameter Isotope lt Isotope name gt Repeat times is greater than 1 See section 6 2 7 Repeat Counting for more details WIN CPM Measured CPM in counting window TOTAL CPM Measured total CPM in the whole spectrum 6 2 4 GLP Criteria For each of the above quantities you can set a low and a high limit by setting the FILES menu item GLP data Operation to Criteria and pressing the ENTER key when the highlight bar is on the menu item Do operation The oth
50. relative spillover and crosstalk values Let us denote this new matrix with the letter M If this matrix is multiplied with the following 1x 2 N vector B 160 10 3 RIA IRMA RATIO assay counting Isotope A activity in detector 1 Isotope B activity in detector 1 Isotope B activity in the last used detector the result is the following 1x 2 N vector A Measured activity in isotope A counting window for detector 1 Measured activity in isotope A counting A window for the last used detector Measured activity in isotope B counting window for detector 1 Measured activity in isotope B counting window for the last used detector Thus A MB Then the matrix M is inverted Let us denote the inverted matrix with M When this is multiplied during assay counting with a vector consisting of the measured CPM values in both counting windows and for all detectors the result vector consists of spillover and crosstalk corrected isotope activities for all detectors that is B M A If the matrix M cannot be inverted the message The spill matrix could not be inverted No spill correction is made is printed If the absolute value of the determinant of the matrix M is smaller than 0 1 the message Results for CPM may be inaccurate is printed 10 3 2 Setting the assay counting window The assay counting window is already known in keV s but it must be converted to MCA channels before the number of counts in it can be
51. replicate tubes The counting window used is the same for all replicates and is the one determined from the sum spectrum The counts values are corrected for dead time background and isotope decay to get corrected CPM 10 2 10Checking detectors for stability if repeat normalization is used The parameter SYSTEM Isotopes lt isotope name gt Repeat times gives the number of times each isotope normalization measurement is repeated The total time a sample is measured in each detector is this number multiplied with SYSTEM Isotopes lt isotope name gt Normalization time The measured counts in repeat measurements are compared with each other and the program calculates the probability that a Poisson distributed random variable having the same mean would have had a greater variance than was observed in these repeat measurements This probability is called Significance level and its unit is If it is often near zero or near 100 this means that there is systematic error in repeat measurements In the extended isotope normalization printout the column heading is SIGNIF LEVEL Since the counting window is determined from the sum spectrum and thus is the same in all test repeat measurements short term gain variations are detected by this measurement The formula used to calculate Significance level is as follows Let CTIME the counting time used in repeat measurement COUNTS the counts measured in the isotope counting window
52. setting the menu item Operation to Criteria in the menu FILES GLP data and by pressing the ENTER key when the highlight bar is on the menu item Do operation The other items in the menu are used to select the isotope and one of the stored values mentioned above In the same menu that is used to set the limit values you can specify that a warning message a graph or both are printed if during GLP TEST measurement some quantity is not within limits The stored GLP TEST normalization data can later be viewed graphically by setting in the menu FILES GLP data the item Operation to View and by pressing the ENTER key when the highlight bar is on the menu item Do operation The other items in the menu are used to select isotope detector if a 1470 instrument has more than one detector installed and one of the stored values mentioned above GLP graphs can later be printed by pressing the digit key 6 while the graph is displayed The graph is sent to the printer that is connected to the WIZARD printer port if the parameter SYSTEM Printout selections Use printer port is Yes The graph is also sent via the WIZARD PC port to MultiCalc if either SYSTEM Operation mode Evaluation is MultiCalc or SYSTEM Printout selections Without buffering to PC is Yes When the graph is sent to MultiCalc special code characters are added to the data so that the graph can be printed through MultiCalc using the printer that is connected to M
53. spillover to 1 129 window is very large the warning Results for CCPM DPM and RATIO may be inaccurate is printed 10 4 2 Calculating corrected CPM values CCPM for unknown samples The counting windows used for unknown samples are the same as those that were used to make up the spillover correction matrix Isotope peaks are not searched for so possible variations in effective gain are not taken into account i e the windows are not shifted once they have been determined Counting window boundaries are determined to a fraction of channel When counts are summed over a counting window counts from the low and high boundary channels are added only in the proportion that the channels are included inside the counting window 174 10 4 Multiple isotope assays MIA The counts sum is corrected for dead time converted to CPM and background activity is subtracted if selected in the MIA protocol If spillover correction has been specified in the MIA protocol these CPM values from all counting windows are used to form for each measured tube a 1xN vector A as described in the preceding paragraph It is multiplied by the spillover correction matrix M to get a vector B of spillover corrected CPM values If decay correction has been selected for this isotope that is if SYSTEM Isotopes isotope name gt Decay correction is Yes and SYSTEM Isotopes lt isotope name gt Assay zero time is not None the CPM values are further correct
54. the first cassette has 1ts correct protocol selection ID and that the appropriate protocol is stored in the instrument For more information about the ID system see section 2 1 5 Put a STOP ID on the last sample rack to be counted or use a STOP rack or a totally empty rack in order to stop the instrument automatically Load racks starting with the right hand conveyor lane Start counting by pressing the START key The instrument will now count all the loaded samples and the WIZARD CPM software will print out the CPM results During counting you can see the live results on the live display as described in section 2 4 1 by selecting the menu item Show CPM results to see the output from WIZARD on the display The counting will stop automatically when a STOP rack is found You can also press the STOP key on the WIZARD keyboard In that case the following text will appear Continue End assay continue End assay Clear conveyor Depending on whether you want the next assay to be counted or all counting to stop select End assay continue or End assay clear conveyor respectively To override the stop instruction and continue counting select Continue 34 CPM operation 3 9 The FILES function Most FILES functions are not available in CPM mode they are in grey they are only available in RiaCalc WIZ however the following three are 3 9 1 Spectra Store spectra this is for information only It tells if spectra can be s
55. these modes 1 3 2 WIZARD controls If you are a first time user of WIZARD you should read Part 2 WIZARD controls before starting to operate the counter This part explains the basic techniques involved in using the built in display and keyboard s It also explains the barcode ID system Once you are familiar with the techniques of using WIZARD you can proceed to actually operate it 1 Introduction 1 3 3 Using WIZARD to get results WIZARD counts samples and if necessary evaluates the results following instructions given in the form of protocols lists of parameters In normal operation these protocols are already set up so all you need to do is follow the instructions on one of the step by step sheets If however it is your responsibility to create or edit protocols then you will find the information you need to help you in the appropriate one of parts described below CPM counting requires only three parameters to be set as described in Part 3 CPM operation RiaCalc WIZ operation is described in Part 4 Each feature of RiaCalc WIZ is described and guidelines are given to help you use these features to achieve the results you want If you are going to be working with MultiCalc running on an external PC then turn to Part 5 This describes those things you need to know in order to use WIZARD and MultiCalc together The use of MultiCalc is described in a separate User Manual which comes with the software Part 6 tells about Isotope and ba
56. this communication protocol set in WIZARD the parameter SYSTEM Operation mode Evaluation to CPM or RiaCalc WIZ and edit the assay protocol in WIZARD You can save the MultiCalc assay protocol on a diskette and then load it into WIZARD Check however that the specified isotope is the right one also after the protocol has been loaded into WIZARD 5 4 5 Using the communication protocol WIZARD As described above this communication protocol is used when resutls are buffered in WIZARD Each buffered assay is deleted only after MultiCalc has acknowledged that it has received it To use this communication protocol set in WIZARD the parameters SYSTEM Operation mode Evaluation to multiCalc and SYSTEM Printout selections Without buffering to PC to No If you are not using a local printer that is connected directly to WIZARD set SYSTEM Printout selections Use printer port to to No 5 4 6 Softkey F5 INSTALL To see what isotopes are available press in MultiCalc the softkeys F1 COUNTER F5 INSTALL You get a list of available isotopes and short instructions on how to specify them in assay protocols If you have just turned on WIZARD you may get the Framing error message at this point In this case press ENTER and F5 INSTALL again 5 4 7 Curve plotting parameters In the case of a RIA or IRMA the following parameters are available 20 X AXIS concentration 21 Y
57. used in normalization must be shown by attaching an appropriate numerical label to the PROTOCOL area when the rack is being used as a normalization rack 2 9 5 Rack number label Use numerical labels 1 99 to select the rack number attach the label in the area marked RACK SPECIAL This is optional and need not be used at all 2 10 Barcode errors If the ID system fails to read the barcode successfully the rack is handled as if there was no barcode If the code is read successfully but that particular protocol does not exist the message Protocol not found appears on the display and printer and the counter stops 24 2 Wizard controls If there are two numerical codes on the same ID clip the one in the RACK SPECIAL area is taken as the rack number and the one in the PROTOCOL area as the protocol number 2 11 Help The WIZARD software includes an extensive context sensitive Help function If at any point while operating WIZARD you are not sure what to do or what a particular function is just press the HELP key on the built in keyboard or Fl on the external keyboard An explanation of the currently selected feature will appear on the display When you have read the help text you can go back to the function about which you requested help by pressing the EXIT key 2 12 Short cut keys Two short cut keys Alt I and Alt E are available on the main menus level Alt I the I meaning internal sets the mode to RiaCalc WIZ and the SY
58. what follows only Printer selection is explained because Display and Result selections are identical to it Note The registered counts in a counting region window are printed out as COUNTS These values do not include background subtraction or any other correction The following corrections are applied to get the final CPM value background correction if background normalization done dead time correction decay correction if selected spillover correction dual label assays The CPM values shown in the live display are uncorrected 4 6 2 Default or customized outputs The program has a default set of output items shown in the sections following which it prints out if further instructions are not given Therefore when you create a new protocol it is not necessary to change the printout format However you can choose a new type of output from a wide range of the possibilities To alter the printout form first select Printer in the protocol and press E A menu will appear on the screen Printout options Printout fields Printout switches Copy settings from a template 48 4 Operation with internal RiaCalc WIZ Each of these selections is described below 4 6 3 Printout options Printout options allow certain types of output to be selected Most of these options are plots but it is also possible to have the protocol listed as well as the protocol identifier When you select this option a list appears with the
59. window and correcting the result for dead time and converting it to CPM after that background activity is subtracted Only the 1480 counter model includes this value in normalization printout Its purpose 1s to provide a reference value for the calculation of MIA RATIO values 144 10 2 Isotope normalization and GLP TEST measurement Although not printed the standard CPM is also calculated by the 1470 program There it is used after a further isotope decay correction to determine relative detector efficiencies if the counter has more than one detector installed 10 2 8 Correcting for isotope decay If SYSTEM Isotopes lt isotope name gt Decay correction is Yes and SYSTEM Isotopes isotope name gt Norm zero time is not None all normalization measurements are corrected for isotope decay If Norm zero time is Start decay correction is done to the start of the normalization The starting time of a normalization is printed in the title line of the isotope normalization printout If the value of Norm zero time is an explicit time decay correction is done to that time The formula used to calculate the decay correction factor is AAte TT e where T4 the time when the measurement was started To a reference time an explicitly given time or the start of isotope normalization At the measurement time A in where T half life of the isotope A When measured counts or CPM are multiplied with the ab
60. 0 01 SCV from reference precision profile average for only unknowns E B 0 01 As above for channel B E 111 Code text from coding and control list LAG o0 LIT Concentration flag all values but only unknowns LAG B 0 111 As above for channel B K 111 BLANK cpm valid after BLANK in coding all values for both ps KB 111 above for channel B ER 111 above for REFER ER B 111 above for channel B L tte 111 previously for TOTAL L B 111 above for channel B TdTSSS 4 111 above for i controls B 111 above for L B HtHteet 111 above for controls B 111 above for LB ESP 11 Programmable ESP B 11 As above for L B STS 0 111 umerical flag number These are 1 OUT 2 gt STD 3 STD 4 amb 5 gt gt STD 6 lt lt STD 7 CV 8 gt CV 9 gt gt CV STS B 44 0 111 umerical flag number channel B GROUP GR 0 11 Group number of multiple UNKN coding REPL RP 0 1 Replicate of individual sample for unknowns SAMPLE SPL 0 111 Sample No includes controls for all values but only unknowns RESP 11 Response error RESP B 11 Response error for channel B ILF 0 11 Dilution factor EMARK HHEHtHHHHEE 0 111 Remark EOA 1 Application sequence OW 111 Row number 1 individual 2 average 3 second average ATE 111 Date LOCK 111 Time LASS 111 E g 1 if CON
61. 04 Oct 1994 15 04 19 Total counting time 600 Nominal gain 1 00 keV channel Main peak 320 0 kev Main peak at nominal gain 335 0 channels Nominal window coverage 80 00 PEAK PEAK RESOL WINDOW keV DECAYED MEASURED STANDARD CHN DEV LOW HIGH ACTIVITY COUNTS CPM 254 7 24 0 11 6 188 2 428 4 0 9997 460344 46227 VALUES SAVED END OF ISOTOPE NORMALIZATION NORMALIZATION OF 4 I 129 04 Oct 1994 15 14 31 Total counting time 600 Nominal gain 1 00 keV channel Main peak 29 0 kev Main peak at nominal gain 29 0 channels PEAK PEAK RESOL WINDOW keV DECAYED MEASURED HORROCKS STANDARD CHN DEV LOW HIGH ACTIVITY COUNTS EFFICIENCY CPM 22 9 21 1 29 8 15 0 35 0 1 0000 417276 84 1 42115 VALUES SAVED END OF ISOTOPE NORMALIZATION MULTI ISOTOPE ASSAY 10 PROTO1 UNKNOWN SAMPLES 04 Oct 1994 15 39 48 Determinant of spillover matrix 0 00245 Results for CCPM DPM and RATIO may be inaccurate Time limit 600 Max count limit 9999999 Low count time 6 Low count limit 10 Replicates 1 Subtract background YES Eliminate spillover YES 173 10 4 Multiple isotope assays MIA Spill determinant Sequence number Sample number Replicate number Elapsed time Counting time Dead time ISOTOPE COUNTS I 125 417441 Co 57 1661 Cr 51 879 I 129 418891 Sequence number Sample number Replicate number Elapsed time Counting time Dead time ISOTOPE COUNTS I 125 3689 Co 57 1621556 Cr 51 10583 I 129 3698 Sequence number Sample number Rep
62. 10 4 Multiple isotope assays MIA RelativeCpmError SQUAREROOT Variance2 CorrectedCpm CorrectedCpm is corrected for dead time and background if background correction is selected but not for spillover crosstalk or isotope decay If CCPM the final corrected CPM in the MIA printout includes spillover corrections the ECPM printed refers to the quantity CPM which does not have this correction If an unknown MIA sample consists of several replicate tubes the error in the replicate average is calculated in the following way For each replicate measurement define Variance3 SQUARE DeadTimeFactor CountsinWindow Let TotalVariance be the sum of Variance3 s for all replicate measurements and let TotalCountingTimelnSeconds be the sum of CountingTimelnSeconds s for all replicate measurements Then RelativeReplicateAverageCpmError SQUAREROOT TotalVariance SQUARE 60 TotalCountingTimelnSeconds SQUARE BackgroundCpmlnWindow RelativeErrorlnBackgroundCpm CorrectedReplicateAverageCpm Background error is added only if background correction has been selected CorrectedReplicateAverageCpm is corrected for dead time and background if background correction is selected but not for spillover or isotope decay One could think that the variance of the replicate average could be calculated by summing the variances of individual replicate sample CPM s and dividing the result by the square of the number of replicates assuming that all repl
63. 2 1 on page 158 It is multiplied by the spillover correction matrix M to get a vector B of spillover corrected CPM values These are printed e For single label assays that correct for crosstalk the CPM values from all used detectors are used to form a 1 xN vector A N is the number of detectors in use as described in Section 10 2 13 on page 147 It is multiplied by the crosstalk correction matrix M to get a vector B of crosstalk corrected CPM values These are printed 163 10 3 RIA IRMA RATIO assay counting e For dual label assays that correct for crosstalk that is for at least one of the isotopes SYSTEM Isotopes lt isotope name gt Crosstalk correction is Yes the CPM values of both counting windows from all used detectors are used to form a 1x 2 N vector A N is the number of detectors in use as described in Section 10 3 1 2 2 on page 159 It is multiplied by the spillover and crosstalk correction matrix M to get a vector B of spillover and crosstalk corrected CPM values These are printed 10 3 4 Bad spectrum During assay counting a spectrum is considered bad if an I 125 coincidence peak is not found or if it is too small or if an unexpected peak is found The I 125 coincidence peak is not checked in dual label assays if for the other isotope SYSTEM Isotopes lt isotope name gt Spectrum type is Many peaks In this case the printed corrected CPM values are 0 If SYSTEM Isotopes lt isotope name gt
64. 3 1 2 2 on page 159 for the case where M 2 The spillover and crosstalk coefficients can be assumed to independent of isotope activities at least as long as they are not very large When samples containing several isotopes are measured in all the detectors simultaneously the inverse of the spillover and crosstalk coefficient matrix can be used to solve the individual isotope activities in each of the samples just as if all the other detectors were empty and only one Isotope present 178 10 5 Appendix Some basic mathematical formulas For typographical reasons the matrices and vectors in this manual are represented as tables 1 e Ay Am and b by 179 11 Installation 181 11 Installation 11 Installation 11 1 Installation procedure Make the installation according to these instructions and complete the Installation Qualification and Operation Qualification IQOQ report form in parallel 11 2 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 Wallac 1470 WIZARD as this allows the best control over the immediate environment Ventilation in the room 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 impo
65. 4 6 21 2 17 7 46 1 1 0000 104479 0 9814 0 28 9 331 AVG 106419 1 0000 0 28 VALUES SAVED END OF ISOTOPE NORMALIZATION Figure 5 1470 normalization printout when SYSTEM Isotope I 129 Counting window is Dynamic and SYSTEM Printout selections Extended norm printout is Yes NORMALIZATION OF 4 I 129 26 Aug 1994 10 11 26 Total counting time 45 Repeat times Nominal gain 1 00 keV channel WINDOW keV DECAYED MEASURED DETECTOR RELATIVE SIGNIF DET LOW HIGH ACTIVITY COUNTS EFFICIENCY ERROR LEVEL 1 26 0 34 0 1 0000 68311 1 0195 0 34 14 797 2 26 0 34 1 0000 68444 1 0215 0 34 91 101 3 26 0 34 0 1 0000 65691 0 9805 0 35 26 005 4 26 0 34 0 1 0000 65559 0 9785 0 35 86 975 5 26 0 34 0 1 0000 67558 1 0082 0 35 89 982 6 26 0 34 0 1 0000 65592 0 9791 0 35 56 103 7 26 0 34 0 1 0000 69318 1 0346 0 34 51 491 8 26 0 34 0 1 0000 64563 0 9637 0 36 56 986 9 26 0 34 0 1 0000 69154 1 0321 0 34 91 725 10 26 0 34 0 1 0000 65850 0 9825 0 35 7 336 1 0 VALUES SAVED END OF ISOTOPE NORMALIZATION Figure 6 1470 normalization printout when SYSTEM Isotope I 129 Counting window is Fixed and SYSTEM Printout selections Extended norm printout is Yes 152 10 2 Isotope normalization and GLP TEST measurement NORMALIZATION OF 4 I 129 26 Aug 1994 10 33 07 Total counting time 15 Detectors not in use 6 7 8 9 10 DECAYED MEASURED DETECTOR RELATIVE DET ACTIVITY COUNTS EFFICIENCY ERROR 1 1 0000 35254 0 9955
66. 9 For measuring this type of tube every second elevator fork must be removed The instrument program must be informed of these changes as described in this chapter The changes involve some SYSTEM parameters that are only used for this purpose 7 82 SYSTEM setting The parameter SYSTEM Active hardware Forks only at odd positions must be set to YES to tell the instrument program that the elevator forks are only at odd detector well positions The leftmost fork must be at detector well position 1 Note Forks only at odd positions is visible only if the instrument has more than one detector installed y p y The parameter SYSTEM Operation mode Ignore even sample pos must also be set to YES when you are measuring large Eppendorf tubes The default setting NO is used when you are measuring normal vials that can occupy every sample position in the rack Note Ignore even sample pos is visible only if the instrument has only 1 detector installed or if SYSTEM Active hardware Forks only at odd positions is set to YES If Ignore even sample pos is set to YES and a sample is detected at an even rack position the conveyor stops and an error message appears A sample is allowed in an even rack position however if the rack is a background or an isotope normalization rack or a GLP TEST measurement rack Note if the parameter SYSTEM Operation mode Ignore if no holder is set to YES then all even or odd sample positions w
67. A D converter The dead time factor is calculated as shown by the following pseudo code The formula for Tmp2 was chosen heuristically to account for the influence of other detectors CountsFromAllDets is the total counts from all detectors that have been connected to the same A D converter as the current detector CountsFromCurrentDet is the total counts from the detector for which we are calculating the dead time factor CountsFromOtherDets CountsFromAllDets CountsFromCurrentDet CountCheckTime 25 E 8 CountConvertTime 475 E 8 DetectorDeadTime 100 E 8 Tmp1 1 CountConvertTime CountsFromAllDets CountingTimelnSecs 5 CountCheckTime Tmp2 CountConvertTime CountsFromOtherDets CountingTimelnSecs Tmp1 CpmFromDet 60 CountsFromCurrentDet CountingTimelnSecs CorrFac1 60 60 ComFromDet Tmp3 Tmp3 CountConvertTime 5 CountCheckTime 2 Tmp2 2 Tmp4 CpmFromDet CorrFac1 CorrFac2 60 60 Tmp4 DetectorDeadTime DeadTimeFactor Tmp4 CorrFac2 CpmFromDet Ifa sample decays very quickly the dead time factor also decreases during measurement This effect has not been taken into account in the above formulas 10 1 4 Converting isotope counting windows from keV s to MCA channels In order to sum background counts over an isotope counting window the window boundaries must be known in terms of MCA channels For each normalized isotope the isotope counting window boundaries in keV s ar
68. C STD 1 or 2 5 if CONC STD 3 STD 2 2 LASS B 111 As above for channel B QODQUBNWOUUO Corrections in CPM are dead time decay background crosstalk and spillover These fields are used in statistical calculations e g a mean or average is calculated This shows the format of the fields The numbers after fields are the default switch settings See section 4 6 9 73 6 Normalization 6 1 Normalization 6 2 GLP test normalization 75 6 1 Normalization 6 Normalization 6 1 Normalization 6 1 1 Principle The two five or ten detectors of WIZARD allow several samples to be measured simultaneously However as each detector has a slightly different efficiency and background value it is necessary to correct the measured sample counts so that the final result is as if every sample had been measured in the same detector In dual label measurements one isotope being measured will often cause counts to be recorded in the second isotope window and vice versa This effect is called spillover and must be corrected for For higher energy isotopes over 200 keV there will also be crosstalk between detectors This means that a sample being measured in one detector will cause counts to be recorded in other adjacent detectors The higher the isotope energy the greater the crosstalk Thus crosstalk must be calculated and corrected for when it occurs The detector system gain and window must be pe
69. C dai 183 11 4 Unpackin piesien dit e tiers 183 11 5 Checking the mains voltage setting enne enne nennen enne 184 11 6 Connecting up the counter and peripherals sssesesssesseeeeeeeeeenenen eene 184 11 7 Switching on WIZARD aii a 186 11 8 Installing MultiCalc 4 iio eicit talk i dnb rd fni ise iae 186 11 9 Functional check i ien pr ip o featured Re eade cespite e rede ebbe 187 12 Declaration of Conformity for CE marking ccossssosssssossssnossnssonsnsnnnsnsnnnene 189 13 A 95 Trademarks RiaCalc is a trademark and MultiCalc and WIZARD are registered trademarks of Wallac Oy IBM and PC DOS are registered trademarks of International Business Machines MS DOS Microsoft and Excel are registered trademarks of Microsoft Corporation DeskJet and LaserJet are trademarks of Hewlett Packard 1 Introduction 1 Introduction 1 Introduction 1 1 Introduction to PerkinElmer Life Sciences Wallac Oy PerkinElmer Life Sciences Wallac Oy is the world s leading manufacturer of automatic gamma counters and has been a pioneer in this field for many years PerkinElmer Life Sciences Wallac Oy has a well founded reputation for technological innovation and excellence in quality both in products and service 1 2 Introduction to 1470 WIZARD WIZARD is the latest and most advanced gamma counter from PerkinElmer Life Sciences Wallac Oy It comes in different shapes and sizes but each is capable of performing similar magic in your labo
70. Display Choice use lt gt or ENTER More Y Copy An existing protocol can be copied so as to create another protocol with the same contents Select Copy and press E Select the protocol to be copied Give the name the new protocol is to have You may also give an ID number Select do copy and press E Rename Select this option to give a new name to a protocol Select the protocol to be renamed from the list of protocols Give it a new name and or ID number Select do rename and press E Delete Select Delete and press E Then select the first protocol to be deleted Next select do delete and press E The protocol and any associated data will then be deleted Recover Deleted protocols can in most cases be recovered by selecting this option They are saved in the protocol index area There is room on the system disk for 100 deleted and active protocols The larger the number of active protocols the fewer deleted protocols can be saved If there is no room the oldest deleted protocol will be permanently removed to make room for a newly created one No protocol with the same name should have been deleted later than the one you want to recover Print With this function you can print the contents of a single protocol or the protocol index according to the selection you make 41 4 Operation with internal RiaCalc WIZ Isotope names are included in the protocol printout For dual label protocols the code numbers
71. EM Isotope lt isotope name gt Counting window is not Fixed STANDARD CPM is calculated from MEASURED COUNTS which is corrected for dead time and background activity and isotope decay It is printed only for the 1480 counter model and is used in multiple isotope assays MIA as the activity of an isotope standard when the RATIO field is calculated RATIO is equal to an isotope s CCPM activity in an unknown sample divided by the corresponding standard sample s activity SIGNIF LEVEL indicates whether measurement results are repeatable During isotope normalization each measurement can be repeated several times to test detector s for stability The parameter SYSTEM Isotope lt isotope name gt Repeat times sets the number of times each isotope normalization measurement is repeated Thus the total time a sample is measured is this number multiplied with SYSTEM Isotope lt isotope name gt Normalization time Repeats are not done for MIA standard normalizations The measured counts in repeat measurements are compared with each other and the program calculates the probability that a Poisson distributed random variable having the same mean would have had a greater variance than was observed in these repeat measurements This probability is called Significance level and its unit is If it is often near zero or near 100 this means that there is systematic error in repeat measurements 155 10 3 RIA IRMA RATIO assay cou
72. Edit protocol miii asas 29 39 Efficiency Efficiency 96 Electrical requirements End transfer lane use Energy range Er Energy resolution eneessesseeenensennensnnnnnennennnnneennenn Environment sese Eppendorf tubes large Fitting algorithm essere Fixed counting window Footprint eese eerte nne nennen Forks only at odd positions 115 format GIS na inva enti t adiens 106 Repeat counting GLP Criteria nur a 80 A ns reinen 35 58 70 80 GLP test label iere bees iria 24 GLP test sample DPM 96 Good Laboratory Practice sese 79 H 196 Hard disk A tette imer 119 Hard Wares een nis 127 SO A iira anaE E EEEE ETEN SaNa 67 Holder Ignore MOI a a Neel apud Horizontal background printout I ID 22 ID reader for rack ID system Ignore even sample pos Ignore if no holder ds ig Installation sseeseeeseeer eene Instrument serial number Internal RiaCalc WIZ sse Isotope parameters essseeeeeeneeen enne Isotope selection ISOTOPES ieee eE reete eit eae esee ei een K Key click selection essen 102 dno 17 L Labels tirada Background Large vials Leaving the editor sse 34 53 Level selection in MultiCalc
73. HALT YES causes halt meaning that the evaluation is suspended after the standard curve until accepted by the user The third choice does not cause halt but uses the stored reference curve for evaluation 4 5 10 Controls If you do not want to use control samples select NO If you select YES then the following parameters appear Field For control samples normally the concentration of each control is selected however it is possible to select a different quantity and to specify whether it applies to individual controls average values or averages of dilution series averages The next three lines allow target values to be set for the three types of controls Low Medium and High followed by the additional controls Control4 Control5 and Control6 In each case you can give a target value for the control the upper or lower 2SD limit or both plus and minus 2SD limits 44 4 Operation with internal RiaCalc WIZ List This parameter lets you specify a list of controls and the number of patients between controls e g lt start of assay gt LOW CONTROL 10 SAMPLES MEDIUM CONTROL 10 SAMPLES You can set up to 12 controls of one kind e g LOW in the list The list should be long enough to allow the evaluation of assays of varying lengths If you want the assay to end with controls you must adjust the number of patients in the list before each assay If you edit this protocol in an external MultiCalc you can put your own codes in t
74. INSTRUMENT MANUAL January 2002 Eum 1470 WIZARD Gamma Counter gt PerkinEimer life sciences 8 cie Wallac 1470 WIZARD Gamma counter For instruments with software version 3 6 Il PerkinE mer life sciences Wallac Oy P O Box 10 FIN 20101 Turku Finland Tel 358 2 2678111 Fax 358 2 2678 357 Website www perkinelmer com lifesciences 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 Sciences Failure to follow these instructions may invalidate your warranty and or impair the safe functioning of your equipment gt PerkinElmer life sciences Contents Contents A EN Introduction to PerkinElmer Life Sciences Wallac OY ccesccessceseessecseeeseeeseeeeceeeeerenseenseenseenseenaes 7 Introduction to 1470 WIZARD 00 0 e E E E E R E E E E E N E 7 Introd tion AAA eerte E A E AE veut 7 WIZARD co ntr lS s e aan saninssunrnessuihten 17 SS en midi o es iat 17 Keyboard m aste RR tee RU An nme atus 17 Display x ecoute datu utet datei att E 18 Live display 2355 ot tt voto tee etie a m ated du aris 19 Prnciple of the ID systenm uc eR e eei e ete E NU EE 22 Fitting ID labels eta tes re iet debeam e 23 Loading racks the right way round sse enne nennen 23 Instr ction labels dae diet
75. In the latter case also the crosstalk of one isotope to an other isotope s counting window in another detector is corrected for e The corrected CPM activity of an isotope standard is determined if isotope normalization is done at the beginning of a 1480 multiple isotope assay MIA It is used to calculate the ratio of an unknown sample s activity to the standard sample s activity 135 10 2 Isotope normalization and GLP TEST measurement ISOTOPE 1 ISOTOPE 14 Max norm dev 50 Warn assay dev 20 Signif cpm per keV 10 Warn assay dev 5 Signif cpm per keV 2 Name I 125 Name Co 58 Comment Iodine Comment Cobalt Normalization time ee Normalization time 60 Repeat times Repeat times 1 Crosstalk correction x0 Decay correction YES Decay correction YES Half life hours 1711 Half life hours 1445 Norm zero time Start Norm zero time Start Assay zero time Start Assay zero time Start Energy range Normal Counting window Dynamic Counting window e keV Peak pos keV 23 Peak pos keV 810 Window coverage 97 Low boundary keV 180 Threshold level 20 High boundary keV 950 Spectrum type I 125 Threshold level 20 Max coinc dev 25 Spectrum type Many peaks Min coinc height 25 Max assay dev 10 E Max assay dev 30 Max norm dev 20 Efficiency 65 GLP test sample DPM 10000 Figure 3 Default isotope par
76. M Isotopes isotope Spectrum type is Single peak or I 125 then the smoothed spectrum is scanned for unexpected peaks Ifthere are channels that are marked as Maximum and the heights of which exceed ThresholdChannelCounts and which are not the main peak or the coincidence peak then these are marked as unexpected If such peaks are found the normalization is aborted The first unexpected peak is included in the diagnostic info printout 10 2 4 2 1 9Improving the accuracy of the peak position The peak channel number is estimated to a fraction of a channel by fitting the smoothed spectrum with a parabola through the isotope main peak and two adjacent channel count values 10 2 4 2 1 10 Calculating the effective gain The effective gain can be calculated by dividing the main peak energy expressed in detector keV s by the peak position expressed in MCA channels First the main peak energy in absolute keV s which is given by SYSTEM Isotope lt isotope name gt Peak pos keV is converted to detector keV s Then it is divided by the observed MCA channel number of the peak Absolute keV s and detector keV s are discussed in Section 10 1 4 on page 132 10 2 4 2 2 Dynamic keV window in MCA channels If SYSTEM Isotope lt isotope name gt Counting window is Dynamic keV the counting window is determined by the parameters SYSTEM Isotope lt isotope name gt Low boundary keV and High boundary keV First the effecti
77. M kinen Quality Assurance Manager 191 192 13 Index 193 13Index A ASCH result files ae een ent 45 Assay repeat times Automatic Operation essere 121 B Background sess 127 HECK Et APNE E Mene IA dE 111 Counting time 98 Print background 98 101 Background label iir tert 24 Background printout essere 100 Barcode siciliana 22 EMON Sies esse 24 Bald Tate DP 105 BKG label u a ct ee 24 C Cable inicial ndesdeviddecesdh iss eserageteds citen cta cdas 185 Channel A amp B dual labe nee 85 Chromium release 121 LB T TR 111 CLOCK isc 33 103 Communication parameters eseseeeeeeeee 105 ConnecGtlotis ai hie i er npe kie ERAT eda Control files COMO Sacra dia Conveyor malfunction ooooonccncnncnnonconnnnnnconcnnnonnnnnncn nono 89 Copy protocol E Copying an 180t0pe i1 iecit rre darian au 97 Counter number sese 102 Counting time B Counting window sese CPM mode ecciesie nennen CPM mode selection CPM operation ccccesccseesseescesecceeceeeeceseeeeeseeeeseaeenes gau nassen mente 33 193 13 Index CPM Protocol nenne Create protocol Crosstalk ici dida Crosstalk correction ooonooncoonnoooncooncconnconnnonncconoconnnonnnos 94 Curve edit halt ue deter tete ee peers 44 Curve plotting E 66 Customi
78. R ERA eR AES UE 120 Flexible sample handlino A ia 120 Multi user ID sd oca 120 Multiple label multi detector counting ener nnns 120 Isotope selection soo ao ie too nbi OE IE IR unas n ROI MPG IUS 121 Multichaunel analyzer oie HEURE ER RU RU iR Ro o Lisa tess 121 Automatic and manual operation sss nennen eren enne enne nennen 121 GEP performance testing aont eoi n ei e PA E Re P E UE 121 Specifications di TP Physical MEA situe Ee ear ee tes 125 Electrical requirements neuen eee rese iier erede 125 Environmental requirements rte RR RE RS REN HORN SE I eere den 125 Sample O cas 125 DAA ia 126 Instrument conti ias 127 Contents 9 7 Electrical safety requirements ainia 127 9 8 Isotopes defined for 1470 WIZARD a 128 10 Calculation methods 0 19 E 10 1 Background normalization cccesccesecsseeseeeseeeseeseceeeeseceecseceaecsaecaaecaeecaeeeaesnseseeeeaeeneesnasenaeenaeenaes 131 10 2 Isotope normalization and GLP TEST measurement oooccconononnnonenonconncnnnonnnonnnonnnnnn ron nn nono eene 135 10 3 RIA IRMA RATIO assay counting nono nono ener entren nnne nennen nnne nnns 157 10 4 Multiple isotope assays MIA sisse enne enne 171 10 5 Appendix Some basic mathematical formulas cccecceseesseesceesceesceeeeesecesecesecaeeeecaeeseeseeeneeenes 177 11 TAO O LOO 11 1 T stallation procedure ICI Rn D Rd SARI RI dtes 183 11 2 inanis P 183 11 3 Electric powere eia
79. Ratio HorrocksEfficiency Q Ratio In GLP TEST normalization Horrocks efficiency is stored as the absolute detector efficiency if it is calculated Otherwise the stored absolute detector efficiency is calculated by dividing the measured CPM of the sample with the value of the parameter SYSTEM Isotopes lt isotope name gt GLP test sample DPM The CPM value is corrected for dead time background activity and isotope decay The parameter GLP test sample DPM is visible whenever Horrocks efficiency is not calculated See Section 10 2 15 on page 149 Ratio Background activity is not taken into account when Horrocks efficiency is calculated its effect is assumed to be negligible References Monidetektorinen gammalaskin A multi detector gamma counter by Tapio Yrj nen Pro gradu work at Turku University 1984 Pages 72 to 74 Standardising 1251 Sources and Determining 1 5I Counting Efficiencies of Well Type Gamma Counting Systems by Donald L Horrocks Clinical Chemistry 21 3 370 375 1975 10 2 4 2 5 Calculation of isotope peak resolution Resolution is defined as the ratio of the half height width of the peak to the energy of the peak It is calculated by fitting a bell shaped Gaussian curve over the peak using the least squares method and calculating the resolution of this curve This is called Zimmermann s method A Gaussian curve with an area A and peak energy x 1s represented by the function 142 10 2 Isotope nor
80. STEM parameter Printout selections Write results to file to Yes Alt E the E meaning external sets the mode to MultiCalc and SYSTEM parameter Printout selections Write results to file to No These short cut keys are available only when SYSTEM parameters can be edited that is when measurement is not occurring 25 3 CPM Operation CPM operation 3 CPM operation 3 1 Introduction The instructions here describe the routine operation of WIZARD when it is running in CPM mode 3 2 Start up 1 Switch on the printer this should already be connected to the counter 2 You can put the data disk named 1470 Datadisk into the WIZARD disk drive The name label should be facing upwards 3 Switch on WIZARD After about 3 min the display will show 1470 Main Menu QUIN PROTOCOL FILES SYSTEM Submenu Show cpm results Show evaluation results Operate conveyor Cpm Press START to measure 4 Check the time and date by selecting SYSTEM mode and DATE If it is not correct then give the correct value See System Time amp Date setting Return to the main display by pressing EXIT 5 Make sure that the counter is in the right mode i e that the text CPM appears near the bottom ofthe screen as shown in the example above If it does not then select System and then the operation mode The mode must be CPM 3 3 Normalization Make sure that WIZARD has been normalized for the isotope you a
81. Select SYSTEM and Active detectors The following display will appear Active detectors Detector 1 ACTIVE Detector 2 ACTIVE Detector 3 ACTIVE Detector 4 ACTIVE Detector 5 ACTIVE 101 7 3 System mode Detector 6 ACTIVE Detector 7 ACTIVE Detector 8 ACTIVE Detector 9 ACTIVE Detector 10 ACTIVE Choice use lt gt keys Use the up or down arrow keys to select the detector you want to activate or deactivate Use the right arrow key to change the detector from ACTIVE to NOT ACTIVE or vice versa Press Exit Select Save changes and exit when you leave the Active detectors mode After changing the detectors active list you must redo all the normalizations you are going to use Detectors that are inactive are excluded from all operations Sample numbers are adjusted accordingly There is no need to rearrange the samples in the racks the program takes care that all samples are measured in the right order The measuring speed however may be considerably slower because the program is not always able to use all active detectors but uses active groups of one two or five detectors 7 3 8 Active hardware 7 3 8 1 Use rack ID reader NO disables the reader function ID information must be entered manually when the instrument is started If the rack ID reader 1s not used background isotope and GLP normalizations can still be done using the conveyor after it has been started manually To do this pr
82. T Defined by the user Corrections in CPM are dead time decay background crosstalk and spillover These fields are used in statistical calculations e g a mean or average is calculated This shows the format of the fields The numbers after fields are the default switch settings See section 4 6 9 Fl Fl B B RE RE T T N N B P R R un 4 un coauaoaugumuguouu 59 5 Operation with external MultiCalc 5 Operation with external MultiCalc 5 Operation with external MultiCalc 5 1 Introduction The instructions here describe the routine operation of WIZARD when it is connected to a PC running MultiCalc software Since there are many ways to connect the counter the following list should not be taken as the only possible In the example a single WIZARD is connected to a PC running MultiCalc laboratory data management software The operation of MultiCalc is explained in the MultiCalc User Manuals Note the MultiCalc communication protocol should be the one designed to work with the version of WIZARD you are using This will be found on the WIZARD program disk and should be copied from there during installation 5 2 Start up 1 Switch on the printer Note the printer must be connected to the PC 2 Make sure that MultiCalc has been installed on your PC If it has not then follow the instructions in the MultiCalc manual To start MultiCalc when the DOS prompt on your PC shows e g C type
83. Yes in RIA IRMA RATIO assays counting windows are not shifted If this parameter is No in RIA IRMA RATIO assays where no isotope has been set to use a fixed counting window the counting windows are shifted if peaks raise above background and possible spillover counts 7 3 4 6 Ignore if no holder If the parameter SYSTEM Operation mode Ignore if no holder is set to YES then all sample positions without a holder are omitted in position counting This means that replicate samples are considered to exist even if there is an uncounted position between and the mean value is calculated normally If this parameter is set to NO then the replicate will be considered to have been in the uncounted position This will then affect the result of the calculation of the mean value because the replicate value will be missing 7 3 5 Background time 7 3 5 1 Background counting time Background counting time can be changed with this parameter 7 3 5 2 Print background You can print out the background values 98 7 3 System mode 7 3 6 Printout selections 7 3 6 1 Extended norm printout YES Print complete report of normalization parameters This printout will include the Horrock s efficiency for 1 125 if that isotope is normalized 7 3 6 2 Use serial printer port Select Yes if you want results to be output to a printer connected to the serial printer port of the counter 7 3 6 3 Use parallel printer port Select YES if you wan
84. ZARD communication protocol the Terminal parameter must be VT 52 6 2 7 Repeat counting During GLP test normalization each measurement can be repeated several times to test each detector for stability The parameter Repeat times in isotope editor in the SYSTEM menu sets the number of times each isotope normalization measurement is repeated Thus the total time a sample is measured in each detector is this number multiplied with the normalization time that is set in the isotope editor The measured counts in repeat measurements are compared with each other and the program calculates the probability that differences between expected and observed counts in these measurements occured just because of statistical variation This probability is called Significance level and its unit is If it is near zero or one this means that there is systematic error in repeat measurements The number stored is transformed from the Chi square probability that is shown in the printout SIGNIF LEVEL so that 5 corresponds to 50 4 to 10 3 to 1 2 to 0 1 1 to 0 01 0 to lt 0 001 6 to 90 7 to 99 8 to 99 9 9 to 99 99 and 10 to gt 99 999 This is to make very small and large probability values stand out more clearly Possible isotope decay is taken into account when Significance level is calculated For each detector the counting window over which counts are summed is the same for all repeat measurements and is determined from the sum spec
85. above the calculated limit value cause the spectrum to be regarded as bad because it has an unexpected peak If in dual label RIA IRMA RATIO assay measurement neither isotope is of the type Many peaks the search for extraneous peaks is made using the sum of the limits for both isotopes In the diagnostic info printout below the title Peaks in smoothed spectrum that exceed are listed the channel number and smoothed spectrum counts of local maxima for which the counts value exceeds the limit value or in the case of dual labelled RIA IRMA RATIO assays the smaller of the two limit values 169 10 4 Multiple isotope assays MIA 10 4 Multiple isotope assays MIA When a rack having an ID clip with a numeric label in the range from 1 to 99 stuck in the area marked PROTOCOL is encountered during automatic measurement WIZARD starts measuring the assay that has this protocol number An optional rack number can be given by placing a numeric code in the RACK SPECIAL area of the ID clip otherwise the first rack is assumed to have the number 1 and this increases by one for each subsequent rack The possible assay types for an assay protocol are RIA IRMA RATIO and MIA The assay type is determined when a protocol is created MIA protocols can only be created on a 1480 counter with RiaCalc WIZ A multiple isotope assay consists of standard samples followed by unknown samples Both standard and unknown samples can comprise several replicate tube
86. ak channel is printed only if SYSTEM Printout selections Extended norm printout is Yes and SYSTEM Isotope lt isotope name gt Counting window is not Fixed PEAK DEV is the relative difference of PEAK CHN from Main peak at nominal gain It is printed only if SYSTEM Printout selections Extended norm printout is Yes and SYSTEM Isotope lt isotope name gt Counting window is not Fixed RESOL is the ratio of the main peak half height width to the main peak energy It is printed only if SYSTEM Printout selections Extended norm printout is Yes and SYSTEM Isotope isotope name gt Counting window is not Fixed WINDOW keV LOW HIGH are equal to SYSTEM Isotope lt isotope name gt Low boundary keV and High boundary keV if SYSTEM Isotope lt isotope name gt Counting window is Dynamic keV or Fixed If Counting window is Dynamic window boundaries are determined during isotope normalization on the basis of the parameters SYSTEM Isotope lt isotope name gt Peak pos keV and SYSTEM Isotope lt isotope name gt Window coverage Window boundaries are printed only if SYSTEM Printout selections Extended norm printout is Yes DECAYED ACTIVITY is the relative activity of the sample compared to a reference time It is printed only if SYSTEM Isotope lt isotope name gt Decay correction is Yes and the parameter SYSTEM Isot
87. ameters for I 125 1470 counter and Co 58 1480 counter They can be printed with the dialog screen SYSTEM Isotopes Print isotope and normalization Efficienc 82 Max det eff dev 5 10 2 2 Summing spectra of repeat measurements IF SYSTEM Isotopes lt isotope name gt Repeat times is greater than 1 the sample is measured in each detector as many times as specified in Repeat times The total counting time for the sample in each detector is in this case Repeat times multiplied by SYSTEM Isotopes lt isotope name gt Normalization time Isotope normalizations that are done at the beginning of a 1480 multiple isotope assay MIA cannot have repeats and in this case the parameter Repeat times is ignored if it is greater than 1 The purpose of repeat measurements is to check detectors for stability In extended normalization printout the field SIGNIF LEVEL gives the probability that differences in counts values in repeat measurements are just random variations If SIGNIF LEVEL is often very near zero or 100 there is reason to suspect that the counter does not operate properly The separate repeat measurement spectra are not needed to calculate any other data except SIGNIF LEVEL Therefore the repeat spectra are summed together and all other normalization results are determined from this sum spectrum This gives better accuracy than for example using only the first repeat measurement spectrum 10 2 3 Summing
88. an make a background normalization whenever you like without repeating the isotope normalization 6 1 6 Manual normalization See the chapter on Manual operation for how to do manual normalization and things to be aware of with it 79 6 2 GLP test normalization 6 2 GLP test normalization 6 2 1 Introduction Instrument performance can be monitored by running GLP test normalizations at regular intervals These store data that can later be viewed in graphical format GLP means Good Laboratory Practice A GLP test normalization is similar to isotope normalization only results are stored differently Data obtained in a GLP test normalization are not used in assay measurements but are tested against preset limits and then stored so that they can later be compared with other test normalizations using the same isotope This comparison is done by presenting the values of some measured parameters as a function of time so that any systematic trends or large random deviations can easily be discerned 6 2 2 GLP test normalization rack A GLP test normalization rack has only one holder and sample which is at the last position of the rack The rack has a clip with the TEST instruction at the RACK SPECIAL position and the isotope code at the PROTOCOL position Counting time is set by the parameter Normalization time which is found in the SYSTEM menu under Isotopes lt Isotope name gt The printout is similar to isotope normalization printout It is
89. art 5 7 3 4 2 Manual mode You can operate WIZARD as an automatic or manual counter with any of these three evaluation modes assuming the appropriate options are installed on your WIZARD The change from automatic to manual is accomplished by selecting Manual mode Yes Note the change to manual mode cannot happen while the cover over the detector block is open This is because the change to manual mode causes the raising of the arm which transports samples from the conveyor to the detector block For safety reasons this movement cannot happen without the cover being closed Afterwards the cover must be opened for accessing the detector block See the chapter on Manual mode for mode details 7 3 4 3 Store assay spectra Assay spectra can be stored in files and later retrieved in text form They can be stored in files in the instrument hard disk Each protocol and run id number 1 99 can have one assay spectra file that stores all spectra measured 97 7 3 System mode during the assay The spectra can later be saved to a datalogger disk or sent to a PC or a mainframe in text format The files can also be deleted You can view or print the stored spectra in graphical format by importing the text file to a spreadsheet program such as Microsoft Excel To enable the storing of assay spectra set the parameter to Yes To send stored files to a PC or to save them on a datalogger disk or to delete them go to the FILES menu item Spectra and se
90. as 65 Editing a MultiCalc protocol sess enne nennen enne 65 E ona 68 The FILES function ds cede atid dete dam aatem ais 69 Real trie clock ie e tat eoa 70 Selectable oUtp ts aet oett quet ide deu oed eut 73 NOFMAlZA ION eT A Normalization 5c A chee ed ee aes elsi eee 77 GLP test normal zation iii eii 80 Additional WIZARD functions usssssssssnsssnsnnssnsnsssnsnnsnnnnnsnnnnnsnsnnnsnsnonsennnnsnnnne OD Dual label count ng tote rr e ra ema iE PD REPE ODE 85 Manual operation oer obi oie ia ao eee lat 89 SMA eb oed piel vedette nue dort 93 STAT counting SR RA NEN MD RU UM IN E S 107 Power failures du cos t Aene e eT NN A EE ON NT 109 Routine maintenance ee eer ERES E UO Ren er NAR TRUE ER 111 Safety and radioactive materials unerserseenseensennsennenenensnensnnnnnnnennennennonnennennonnennsennnennnnnenn 113 Large Eppendort tubes esc RO RN NN ENTORNO te 115 Instrument description iocus reni leisen L19 A RR 119 Self containied ree RE OE RAS RU eA eee icto tu ee O 119 Interactive control cesen ne 119 Externalicommunication ioco ei E EEEE EE E E pe tu R E en 119 Multi technology data management software u ccesessesssnsnensnnennennnenneenneennennse nennen nennen nenn 119 Customization and upgradability 4 oe ERR ec s P RE Ere Urea ee ise 119 COMPACIHESS P e 119 High efficiency detectors ON 120 Detector shielding A I
91. asure each individual replicate tube The counting times can be different if counting is terminated because the max counts limit has been reached or if the tube has been rejected because of low activity The replicate averages are printed if this has been specified in the MIA protocol 10 4 5 TOTAL CPS and TOTAL DPS These two values are stored in the file TOTALCPS TXT in the root directory of the instrument hard disk TOTAL CPS is the sum of all printed corrected CPM values of measured unknown tubes in the assay These CPM values are corrected for dead time for background if selected in MIA protocol for spillover if selected in MIA protocol and for isotope decay if selected in isotope parameters The DPS value is obtained by dividing the printed corrected CPM value by the actual coverage of the isotope counting window and by SYSTEM Isotope lt isotope name gt Efficiency TOTAL DPS is the sum of all DPS values of unknown tubes in the assay 10 4 6 How CPM error is calculated when 1480 MIA unknown samples are measured In MIA printout this quantity has the title ECPM The following formulae are used Variance1 SQUARE DeadTimeFactor 60 CountingTimelnSeconds CountsInWindow RelativeErrorlnBackgroundCpm 1 0 SQUAREROOT BackgroundCountsInWindow Variance2 Variance1 SQUARE BackgroundCpmInWindow RelativeErrorlnBackgroundCpm Background error is added only if background correction has been selected 175
92. at the instrument reads the ID labels correctly Compare the results obtained with those on the final test data sheet from PerkinElmer Life Sciences Wallac Oy Send the printout to PerkinElmer Life Sciences Wallac Oy with the IQOQ report Finish the test by changing Extended norm printout back to No 187 11 Installation 188 12 Declaration of Conformity for CE marking 189 190 1390 3693 J DECLARATION OF CONFORMITY FOR CE MARKING PerkinEime INSTRUMENTS 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 1470 Wizard Gamma Counter Valid from serial number 4700909 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 Date and place of issue 01 June 2001 TURKU FINLAND Name and signature or equivalent marking of authorized person Pete AGE Pekka
93. aved or not and if they are to be sent to an output device or not The actual settings can be made in SYSTEM Operation mode Store assay spectra See there for more details Operation this allows you to handle the files of spectrum data The options are Delete delete the spectrum file data Save to disk save the spectrum information on the program disk in WIZARD Send to PC send the spectrum information to a PC that is connected to WIZARD 3 92 GLP data This allows you to handle the GLP data obtained in GLP test normalization There are four options some of which lead to other options View allows you to view the GLP data Delete delete GLP data Criteria allows you to select the warning limits for different types of GLP data You can select Isotope you can select the isotope type from those for which a GLP test normalization has been enabled Item many items appear from which you can select the one which you want the GLP data PEAK BGRD EFFIC RESOL EFFICIENCY COVERAGE CHI PROB WIN CPM TOTAL CPM Print criteria select the isotope for which you want the criteria to be printed out 3 9 3 Waste log file A waste log file that contains the total CPS and DPS values of all measured assays isotope normalizations and GLP TEST measurements can be printed or stored on a datalogger disk The file can also be deleted or sent via the PC port to an external PC The waste log can contain approximately 700 entries after th
94. aving one 3 5 disk drive and a 30 Mbytes hard disk A warning is issued when the instrument harddisk becomes full If at the beginning of an assay or normalization there is less than 3 Mb free disk space in the instrument hard disk an error message is displayed and printed and the conveyor is cleared The computer contains 1 Mbyte of RAM memory two serial RS232C ports one parallel port and a built in 9 screen monitor There is a built in keyboard and an optional additional full PC keyboard 8 3 Interactive control The built in display can be used in conjunction with the keyboard for interactive control of the counter In addition it can display information about the state of counting providing either live numerical or graphical information The built in keyboard has pressure sensitive keys with tactile feedback Most operations can be set up and controlled with this keyboard which means that you do not need to have a PC near the counter 8 4 External communication WIZARD can also be controlled via an external computer and can send results via a serial RS 232C interface to external devices such as a printer or PC or via MultiCalc to a LAN or mainframe 8 5 Multi technology data management software WIZARD can be used to deal with all gamma counting as part of an integrated label measurement system by using the Wallac MultiCalc software package which runs on an external PC This software not only supports gamma counting but all other technolog
95. ays used so expected peak position has no meaning there Low channel limit and High channel limit define the interval inside which the isotope main peak should be They are obtained with the following formulae LowChannelLimit ExpectedPeakChn 1 0 MaxPeakDev 100 HighChannelLimit ExpectedPeakChn 1 0 MaxPeakDev 100 In isotope normalization MaxPeakDev is equal to SYSTEM Isotope lt isotope name gt Max norm dev In RIA IRMA RATIO assays MaxPeakDev is equal to SYSTEM Isotope lt isotope name gt Max assay dev The highest isotope peak between LowChannelLimit and HighChannelLimit is assumed to be the isotope main peak If an isotope has been set up to use fixed windows there is no need to determine the isotope main peak channel number and hence neither LowChannelLimit or HighChannelLimit Used peak channel is the highest isotope peak between LowChannelLimit and HighChannelLimit It is assumed to be the isotope main peak channel number provided that such a peak exists and it is sufficiently high see Sections 10 3 2 2 1 on page 162 and 10 3 2 2 2 on page 163 If no such peak exists Used peak channel is set equal to Expected peak chn In both cases the counting windows determined during isotope normalization are shifted by the same relative amount that Used peak channel differs from the main peak channel determined during isotope normalization Peak begin channel and Peak end channel ar
96. boundary keV to a value higher than the MCA high limit e g to 3000 keV you get an open counting window that includes all counts registered by the detector 10 2 4 1 2 Converting keV s to MCA channels If the ratio of keV and the corresponding MCA channel number were constant one could convert keV s to channels with the formula keV EffectiveGain However this is complicated by the fact that the energy scale of the detector is not linear between 31 and 662 keV We can take this into account by introducing so called absolute keV s and detector keV s When an energy value is converted from keV s to channels it is done in two steps first it is converted from absolute keV s to detector keV s Then it 1s divided by the effective gain to get a MCA channel number ChannelNumber Conversion from a MCA channel number to keV s is done in the reverse manner first the channel number is multiplied by the effective gain to get a detector keV value this 1s in turn converted to an absolute keV value The function to convert from absolute keV s to detector keV s is the following We use pseudo code 157 10 2 Isotope normalization and GLP TEST measurement IF Absolute_keV lt 31 0 THEN Detector_keV Absolute_keV ELSE IF Absolute_keV lt 122 0 THEN Detector_keV Absolute_keV 3 875 0 875 ELSE IF Absolute_keV lt 320 THEN Detector_keV Absolute_keV 11 65 0 99 ELSE IF Absolute_keV lt 662 THEN Detector_keV Absolu
97. ckground Normalization This is an operation which has to be done before WIZARD is used to count samples with a particular isotope When it has been done once it should be repeated occasionally e g after half a year Part 6 also tells about how to do performance testing with GLP test normalization Part 7 describes a number of functions which are available in addition to the three main ways of using WIZARD described in parts 3 4 and 5 Part 8 of this manual gives you a description of how WIZARD works You do not need this information for normal operation but it will help you to have confidence in your results when you know how WIZARD has been designed to give you the very best Detailed specifications are described in Part 9 giving numerical values for e g efficiency background etc In Part 10 there is also a description of the calculation methods used in WIZARD Part 11 contains the information you need when installing WIZARD for the first time Normally this will be done by a service engineer so you will not need this information Part 12 contains the alphabetical index to this manual CPM operation of WIZARD 4 Fix ID clips to racks Fix ID clip here P2 ID labels barcodes are stuck to an ID clip which fits onto a rack to tell WIZARD the function of the rack A counting protocol is a set of three parameters time max counts limit and isotope which control counting Rack number is optional and allows each rack to have its own
98. counting efficiency see the specifications for actual values This design breakthrough has been made possible by means of a unique sample changer mechanism which takes the ten samples arranged linearly in a rack and moves them to the detector wells These are arranged in a staggered configuration which maximizes the shielding between detectors while minimizing the size of the detector block 8 9 Detector shielding The detector assembly is surrounded by a minimum of 12 mm of lead shielding against radiation in the vertical plane The shielding against radiation from samples on the conveyor is 30 mm 1 1 4 The shielding between detectors is 7 mm 8 10 Flexible sample handling Samples are carried in racks which are made of plastic with place for 10 samples rack They are provided with individual sample carriers which can be replaced in case of contamination These racks and sample carriers can be used as they are in a centrifuge This sample carrier system enables WIZARD in automatic mode to count sample vials of any shape up to 13 mm in diameter or microspheres which need not even be in vials In manual mode you can count sample vials up to 17 mm 8 11 Multi user ID system WIZARD is designed for automatic multi user operation Each rack can be provided with two barcodes each having capacity for 2 digits or a special code word These barcodes are used to tell WIZARD how to count the samples Each user can load samples with racks coded f
99. ction and continue counting select Continue 4 9 The FILES function 4 9 1 Introduction RiaCalc WIZ can produce different types of files This function in the main menu called FILES allows you to perform operations on and with these files In CPM mode these files are not produced and in the external MultiCalc mode the same files are handled by MultiCalc on the external computer not on WIZARD There are five types of files which can be handled with this function Input files Standard curves Results files Controls Trends 53 4 Operation with internal RiaCalc WIZ In order for any of these files to be produced you must select the ones you want You do this with the Saved files parameter in a protocol see section 4 5 11 4 9 2 File operations submenu When you have selected a file a file operations submenu will appear This includes the following items for each file type Operation This shows the currently selected operation If you want a different operation then highlight it and press E A list of alternative operations will appear The actual files operation available for each file type are described in later sections The actual operation you select will determine what the remaining lines of the file operations submenu are In every case at least the following lines will appear Protocol Selects the protocol to which the file to be operated on belongs Run id Specifies the run id number of the file that is to be operate
100. ction to select a marker which you can then move with the arrow keys to mark any position in the spectrum In this way you can find out the exact position of any peak WinLo and WinHi These allow you to set an upper and lower window limit marker so that you can obtain the counts within the window You can also print out the spectrum if you press Print screen on the external keyboard provided CAPS LOCK is not on and there is a printer connected Spectra can also be printed via MultiCalc to the printer that is connected to the PC running MultiCalc To do this the following conditions must be met If the instrument is in MultiCalc mode then MultiCalc itself must be in online mode when the Print Screen key is pressed If the instrument is in Cpm or RiaCalc WIZ mode then the instrument parameter SYSTEM Printout options Without buffering PC must be YES and MultiCalc must be receiving data from the counter 21 2 Wizard controls Spectra are printed correctly only if in the WIZARD communication protocol the Terminal parameter is VT 52 Note to change quickly from the spectrum of one detector to another 1 10 just press the respective number key 1 0 on the keyboard You can also use the and keys to scroll through the numbers 2 5 Principle of the ID system WIZARD is an automatic gamma counter This means that samples from several assays can be loaded onto the conveyor and WIZARD can be left to count them by itself To
101. cy has not changed between the two isotope normalizations The following 2x2 matrix is set up for each detector that is in use CPM in isotope A spectrum CPM in isotope B spectrum using the counting window of using the counting window of isotope A isotope A CPM in isotope A spectrum CPM in isotope B spectrum using the counting window of using the counting window of isotope B isotope B Then values in each column are then divided by the diagonal value in that column This converts measured activities to relative spillover values Let us denote this new matrix with the letter M If this matrix is multiplied with the following vector B Isotope A activity in a sample Isotope B activity in a sample Measured activity in isotope A counting window A 23 Measured activity in isotope B counting window Thus A MB Then for each detector the matrix M is inverted Let us denote the inverted matrix with M When this is multiplied during assay counting with a vector consisting of the measured CPM values in both counting windows the resulting vector consists of spillover corrected isotope activities that is B M A If the matrix M cannot be inverted the message The spill matrix could not be inverted No spill correction is made is printed If the absolute value of the determinant of the matrix M is smaller than 0 1 the message Results for CPM may be inaccurate is printed the result is the following vector A 10
102. d how wide it is Since we know the energy of the peak and the nominal gain we can calculate the approximate channel number of the peak To filter out random and systematic errors in channel count values the spectrum is first smoothed 10 2 4 2 1 1Spectrum smoothing The smoothing is such that peaks with half height widths between 10 and 100 channels get almost monotonous slopes It consists of two parts We denote the counts in channel N with CTS N In the first part the counts value of a channel N is replaced by CTS N 2 CTS N 1 2 CTS N 2 CTS N 1 CTS N 2 CTS N 3 8 If a channel number in the above formula would fall outside the range from 0 to 1023 the counts are taken to be 0 In the second part the modified counts value of a channel N is further replaced by CTS N 4 CTS N 3 CTS N 2 CTS N 1 CTS N CTS N 1 CTS N 2 CTS N 3 8 138 10 2 Isotope normalization and GLP TEST measurement Note that all window counts and CPM values are later calculated by summing counts from the original unsmoothed spectrum The smoothed spectrum is only used to determine the isotope main peak channel number and to make a qualitative assessment of the spectrum e g whether the spectrum contains unexpected peaks 10 2 4 2 1 2Marking peaks Peaks are sought in the smoothed spectrum by sliding a five channels wide window over the spectrum The counts values in the two channels on both sides of the centre channel ar
103. d isotope main peak was used to shift the counting window determined at isotope normalization 168 10 3 RIA IRMA RATIO assay counting Peaks in smoothed spectrum that exceed This line in the diagnostic info printout is used to show some prominent peaks in the spectrum The limit above which peaks are shown is obtained in the following way Using the parameters SYSTEM Isotopes lt isotope name gt Signif cpm per keV and SYSTEM Isotopes lt isotope name gt Threshold level first convert the former parameter to a counts value and add to it the fraction given by the latter parameter of the height of the isotope main peak in the smoothed spectrum This sum is the limit value In dual label RIA IRMA RATIO assays the limit is the smaller of the two limits The parameter Signif cpm per keV is used to indicate the amount of background activity at the energy of the isotope main peak Local maxima that are smaller than this are not considered eligible for being the isotope main peak Once the isotope main peak has been found it is assumed that stray counts due e g to Compton scattering are raising the background activity level outside the main peak by the fraction given by Threshold level If in isotope normalization and in single label RIA IRMA RATIO assay measurement the SYSTEM Isotopes lt isotope name gt Spectrum type is Single peak or I 125 for the isotope used any extraneous peaks in the spectrum that rise
104. d on This run id is selected from a list of free ids and is not entered with the numeric keys This is to prevent duplication of run ids Do operation The submenu for all operations end with this line Selecting it and pressing E actually starts the operation defined in the previous lines If this line is displayed in subdued colour it means that the selected operation cannot be done You can also do the operation by first pressing the EXIT key and then selecting the menu item Do specified operation or alternatively by pressing the Ctrl S key In these cases the menu selection bar can be on any menu item To return to the main menu without doing the operation first press the EXIT key and then select the menu item Quit do not operate or alternatively press the Ctrl C key You can return to the main menu directly by pressing only the EXIT key if you have invoked the Do operation and the parameters that specify the operation have not changed after that 4 9 3 Input file This comprises data which can be evaluated by RiaCalc WIZ This data can be either previous output from RiaCalc WIZ or data entered by the user The main point is that it is data in a format acceptable to RiaCalc WIZ The functions available for input files are Create Creates a new file and lets you edit it Edit Lets you edit a file Copy Lets you copy a file Print Lets you print a file Evaluate Lets you evaluate a data file Delete Lets you delete a file If
105. d plots are Left right arrows for selecting the value to be edited Up down arrows for defining how many steps to move each time the left right arrow is pressed WinL 1 Press 1 to set the left limit of a window WinR 2 Press 2 to set the right limit of a window Calc 3 Press 3 to recalculate values after making changes to a plot Del Undel 4 Press 4 to delete a point The mark will change to a square and the point will not be included in calculations Pressing 4 when the cursor is on a deleted point will cause the point to be treated again as a normal point Era 5 Pressing 5 allows points within a window to be permanently removed They cannot be returned Print 6 Press 6 to printout the current plot When you exit from plot editing done by pressing E the changes you have made will be saved 4 9 9 Spectra Store spectra this is for information only It tells if spectra can be saved or not and if they are to be sent to an output device or not The actual settings can be made in SYSTEM Operation mode Store assay spectra See there for more details Operation this allows you to handle the files of spectrum data The options are Delete delete the spectrum file data Save to disk save the spectrum information on the program disk in WIZARD 57 4 Operation with internal RiaCalc WIZ Send to PC send the spectrum information to a PC that is connected to WIZARD 4 9 10 GLP data This allows you to handle the GLP data ob
106. ded in the counting window and so also photons with energies exceeding the MCA high limit are counted Threshold level Specifies the height under which peaks other than the one given by the parameter Peak pos keV are considered insignificant This value is expressed as a fraction of the height of this main photopeak 95 7 3 System mode Spectrum type There are three possibilities Single peak this applies to most isotopes those which have only a single photopeak in the range of the multichannel analyzer 1 125 this is only for the I 125 isotope having a distinguished coincidence peak Many peaks for isotopes having several photopeaks Compton or other secondary peaks Maximum coincidence deviation This parameter gives the maximum allowed deviation of the 1 125 coincidence peak with respect to its expected position which is twice the channel number of the I 125 primary peak Minimum coincidence height This parameter gives the minimum required height of the I 125 coincidence peak as a fraction of the height of the I 125 primary peak If the coincidence peak is smaller than this limit a Bad spectrum message is printed Maximum assay deviation This is the maximum allowed deviation of the isotope peak from its expected position in an assay measurement If the deviation exceeds this value the Bad spectrum message is printed Maximum normalization deviation This is the maximum allowed deviation of the isot
107. default selection as follows Protocol id number YES Protocol NO Standards in table format YES Std curve in graphical format YES Comparison curve NO Controls in table format NO Controls in graphical format NO Response error relationship NO Prec prof in table format NO Precision prof in graphicalformat NO Trends NO Histogram NO Protocol id number prints the protocol name and date and Protocol prints the protocol contents Results for standards can be output as curves and or tables of values Results for controls can be output as plots or tables of values where the most recent control value is the first item in the table The age of the results increases as you go down the items in the table Trend values are output as plots Precision profile output can be as tables or plots A plot of the response error relationship and a histogram of results are also selectable Select YES or NO by pressing the left or right arrow keys Press EXIT when ready 4 6 4 Printout fields Printout fields comprises numeric values and flags These are printed for individual samples unlike printout options which are printed for a complete assay Select this option and you will see a list of the default settings Left margin of paper or display SEQ sequence or tube number PAT patient number for unknwns CODE type in coding or contr list TIME counting time in seconds CPM counts per minute chn A CONC concentration chn A
108. during repeat measurement i 146 10 2 Isotope normalization and GLP TEST measurement DCF the factor that when multiplied with the measured counts gives isotope decay corrected counts in repeat measurement Then define ECOUNTS FOE ESTIMATE CTIME i DCF COUNTS ESTIMATE CHI A AAA ESTIMATE The Significance level is the value of the Chi square probability function with the argument equal to gt CHI and the degrees of freedom equal to one less than the number of repeats References Monidetektorinen gammalaskin A multi detector gamma counter by Tapio Yrj nen Pro gradu work at Turku University 1984 Pages 74 to 78 Numerical Recipes in C by William H Press Brian P Flannery Saul A Teukolsky William T Vetterling Cambridge Cambridge University Press copyright 1988 ISBN 0 521 35465 X The algorithm for calculating the Chi Square probability function is based on the one given in the book referred to above pages 488 and 171 to 178 10 2 11 Relative detector efficiency Relative detector efficiency values are determined by correcting standard sample CPM values for isotope decay and dividing the value for each detector with the average of all detectors that are in use See Section 10 2 7 on page 144 and Section 10 2 9 on page 146 10 2 12Storing normalization sample spectra to enable spillover correction During isotope normalization the normalization sample spectrum is stored for each detector
109. e Diagnostic info printout those peak channel numbers are printed whose smoothed spectrum height exceeds this new ThresholdChannelCounts See Section 10 3 8 on page 164 If SYSTEM Isotopes lt isotope gt Spectrum type is Single peak or I 125 then the smoothed spectrum is scanned for unexpected peaks If there are channels that are marked as Maximum and the channel counts of which exceed ThresholdChannelCounts and which are not the main peak or the coincidence peak then these are marked as unexpected If such peaks are found the spectrum is considered bad and measured counts are set to 0 in the assay printout The possible first unexpected peak is included in the diagnostic info printout 162 10 3 RIA IRMA RATIO assay counting 10 3 2 2 2 Dual label assay First isotope main peaks are determined separately for both isotopes in the order the isotopes are in the assay protocol This is done as explained in the previous paragraph If both isotopes would have the same peak as the main peak this means that for them both the intervals where the highest most active peak is assumed to be the main peak overlap and the same peak is the highest one in both of them In this case a dividing line is set at the geometric mean of the expected peak positions The highest most active peak is assumed to belong to that isotope on whose side of the dividing line it lies The geometric mean of two numbers a and b is equal to yab If the highest isoto
110. e ate PR ar 24 Numerical labels 5 tee ced nuntia te ete 24 Barcode Error dato voe ee iat done e t ars 24 Help coats td amitteret Ned e Lp Es PM tI AA IDE eiusd DL BI DIU AR 25 Shortcut Keyss es ate iste leto dl e ttl tt Ao e e taco al e a Pte 25 Ad M 29 Introduction ii eat 29 NN NN 29 Normalization uet e mine eden ea as cas Raven SS 29 Protocol editing ast kun said dani 29 Parameters available itd eR dt habitats 33 oin EE 33 Eecaving the editor iui aute dep A UI o ee d eure 34 RUNNING n assay sso eade dae Re os n aede as dta den ee ud 34 The EIEES fu nctiOn tie tet oe HERO a Qe diem e ib aive 35 Operation with internal RiaCalc WIZ 4 eeeeeee etes seen eere senno se ensseeessees 39 Introduction z 5i et deem I AS EN EN REDE 39 Start UP ace asthe cass fads e eR ER C CR Er REO EI RENE QUERN eer RR 39 Normalization u eec eue esie e eR HERE I E ORDER iaa 39 Protocol editing eese edant tani Wt i ee de e RR icis 39 Parameters available isse Rh RH ER I E A 43 Output editing ues eec e RR e RR pco 48 Leaving the editor ic enn edet eie e eI 53 Running ati ASSAY iie eee A e TRE DU RR E RR ecd 53 The FILES functions 53 Selectable outputs ee e a 59 Contents Operation with external MultiCalc esssssossssoossssonssssonssssnnssnsnnssnsnnnsnsnnnsssnnnsnnnee OF Introduction dot taies dere ted a tod nenn a IRR OI WI ER E 63 SLA UP PER 63 Multi alc protocol e e aldo ao ae
111. e compared to determine whether the centre channel is a local maximum or minimum Later whenever the program needs to find peaks within a certain channel interval it only looks at those channels that are local maxima The conditions for a local minimum or maximum are the following Denote with CS N smoothed spectrum counts at channel N Each extremum type requires that both the listed conditions are true Extremum type First condition Second condition Maximum at N CS N 1 lt CS N AND CS N gt CS N 1 CS N 2 lt CS N 1 AND CS N 1 gt CS N 2 Maximum atN CS N 1 lt CS N AND CS N gt CS N 1 CSIN 2 lt CS N AND CS N gt CS N 2 Small max atN CS N 1 lt CS N AND CS N gt CS N 1 Minimum at N CSIN 1 gt CSINI AND CSIN lt CS N 1 CSIN 2 gt CS N 1 AND CS N 1 lt CS N 2 Minimum at N CSIN 1 gt CS NI AND CSIN lt CS N 1 CSIN 2 gt CS N AND CS N lt CS N 2 10 2 4 2 1 3The interval where the peak is expected to be In order to find the isotope main peak the program first defines an interval within which the main peak is assumed to be the highest most active peak This is done with the parameters SYSTEM Isotope lt isotope name gt Peak pos keV and SYSTEM Isotope lt isotope name gt Max norm dev First Peak pos keV is converted to MCA channels This is done as explained in Section 10 2 4 1 2 on page 137 Here nominal gain
112. e counting If however the counting time expires first it will terminate counting even though the max counts limit has not been reached The maximum and default value for this parameter is 99999999 4 5 4 Labels The default selection is single label I 125 If you want a different isotope press E then specify if you want dual label or not then select the isotope s from the list of isotopes see chapter 8 2 Specifications To change the contents of this isotope list go to the SYSTEM mode and the Isotopes parameter see charpter 7 3 in this manual If you want to do dual label counting you must also select the second isotope with the right arrow key or press E to get the label selection menu 4 5 5 X axis conc Choose the scale of the X axis of the standard curve The choices are linear logarithmic or linear logarithmic i e linear fitting and logarithmic plotting Logarithmic scales are commonly used 4 5 6 Y axis resp Choose the scale of the Y axis from the List CPM counts per minute B bound CPM B blank B BO CPM BLANK BLANK REFER LOGIT ln r 1 r r B BO LOG B lg CPM BLANK B T CPM BLANK TOTAL BLANK BO B inverse of B BO T B inverse of B T B F r TOTAL r r CPM BLANK F B inverse of B F PROGR customized response PROG LOG PROG with logarithmic coordinates 43 4 Operation with internal RiaCalc WIZ In this list B bound is the response value BO is the reference
113. e known If SYSTEM Isotope lt isotope name gt Counting window is Dynamic they are determined during isotope normalization otherwise they are given by the parameters SYSTEM Isotopes lt isotope name gt Low boundary keV and High boundary keV These can be converted to MCA channels if the gain keV channels is known If Counting window is Dynamic or Dynamic keV then the effective gain is calculated by dividing the energy of the main isotope peak in keV s by the actual position of the main isotope peak in MCA channels The program has stored the position of the main isotope peak in MCA channels the last time an assay was measured using this isotope or when the isotope was normalized if no assays using this isotope have been measured since normalization If Counting window is Fixed then nominal gain is used when this conversion is done For 1470 and 1480 normal energy range the nominal gain is equal by the parameter SYSTEM Active hardware Nominal gain The factory setting for it is 1 0 keV channel If the value of this parameter is changed it affects only how keV s are converted to channels and vice versa by the software not the effective gain of the electronics circuit The latter must be set by a service technician For the 1480 model the parameter Nominal gain is the normal energy range gain The nominal gain for the 1480 extended energy range gain is 2 0 keV channel and cannot be changed 132
114. e major parameters in the protocol are mentioned below They are described in detail in the MultiCalc User Manual under Protocol parameter setting 5 4 1 Single Dual label selection 01 DUAL LABEL Select YES if you want to count dual label samples otherwise select NO 5 4 2 Length of counting 02 COUNTING TIME MAX COUNTS Give the counting time in seconds and optionally the cut off value for the number of counts accumulated A larger count value than this number stops the counting even if the counting time limit has not been reached 5 4 3 Isotope selection 03 MEASURING PARAMETERS This is where you select the isotope and the parameters which define measurement using that isotope When you select parameter line 3 three softkeys appear Fl I 125 F2 60557 F3 1 o0 F1 and F2 correspond to isotope numbers 1 and 2 F3 is for dual label counting with iodine and cobalt Any other isotope must be selected by typing the number see the default list in chapter 9 Specifications but also the following section about limitations 5 4 4 Available isotopes The used isotope s are given in the MultiCalc assay protocol parameter 03 MEASUREMENT PARAMETERS We call this parameter here isotope number Its range is 1 99 and it can be interpreted either as one isotope code number in the range 1 99 or as two concatenated isotope code numbers in the ranges 1 9 and 1 10 65 5 Operation with external MultiCalc If you set DEFINE WIZARD
115. e the nearest local minima around Found peak channel Resol start chn and Resol end chn give the interval within which the peak is over 67 of its maximum height in the smoothed spectrum During isotope normalization the program fits a bell shaped Gaussian curve over this area and gives the calculated resolution of the fitted curve as the peak resolution See Section 10 2 4 2 5 on page 142 This peak has noise is printed if the smoothed spectrum between Peak begin channel and Found peak channel and again between Found peak channel and Peak end channel is not monotonous See Section 10 2 4 2 1 4 on page 139 This is abnormal peak is printed during isotope normalization if the smoothed spectrum counts at channels Peak begin channel or Peak end channel are over 67 of the height of the peak in the smoothed spectrum It is also printed during a RIA IRMA RATIO assay if the nearest local maximum or minimum to Found peak channel is a local maximum Actual coverage is the ratio of measured counts in a window to Open window counts Neither of these counts values is corrected in any way e g for dead time or for background The measured counts in a window has in the isotope normalization printout the title MEASURED COUNTS If SYSTEM Isotopes lt isotope name gt Counting window is Dynamic then Actual coverage is always slightly greater than SYSTEM Isotopes lt isotope name gt Window coverage Window shifted is printed if the observe
116. e which the flag should appear The flag is CV If you select Not used then the flag is disabled Unkn multipl factor The value you enter for this option will be used to multiply the unknown response values The default value is 1 You can select Not used for this Unkn outlier limit Give a CV value Any unknown which has a CV exceeding this value is not included in the precision profile You can select Not used for this 47 4 Operation with internal RiaCalc WIZ Trends If you want trends to be printed they should be selected here The possibilities are Slope at ed 50 Y intercept Estimated concentration at 20 binding Estimated concentration at 50 binding Estimated concentration at 80 binding Blank over Total ratio Reference over Total ratio Blank over Reference ratio Minimum detectable concentration Parallelism factor Difference slope Histogram The default setting for each is NO 4 5 17 Factors You can set the values for the factors UNIT and UNIT_B to show what the units of the results are 4 6 Output editing 4 6 1 Output media The protocol lines Printer Display and Results allow you to specify which results are to be printed on the paper or displayed on the screen or saved as an ASCII file respectively Output selection is specific for each counting protocol The options are the same for each output medium only the medium itself printout device built in display or disk is different In
117. eErrorInRelativeDetectorEfficiency The coefficients have been empirically determined using Poisson distributed random numbers Figure 4 Calculation of relative error in relative detector efficiency If an isotope is normalized at the beginning of a 1480 multiple isotope assay and this isotope has more than one standard replicate the measured spectra of these replicate tubes are summed together and the sum spectrum is handled as if had resulted from the measurement of a single tube Only when the decay correction factor for this sum spectrum is calculated it is taken into account that this spectrum has been obtained from several separate measurements This does not affect the error calculation routines however The individual replicate tube CPM s that are printed at the beginning of normalization printout are determined from each replicate tube s own spectrum using the counting window that has been determined from the sum spectrum see Section 10 2 9 on page 146 If an isotope normalization measurement for each detector is repeated more than once in order to test the detector for stability the resulting spectra are handled much in the same way as described in the preceding paragraph The repeat measurement spectra are summed together and the sum spectrum is handled as if it had resulted from a single measurement But when the decay correction factor for this sum spectrum is calculated it is taken into account that this spectrum has been obtained f
118. each isotope used Wait until the report for each normalization has been printed out 7 2 8 Manual background normalization Select manual mode if that is not already selected Select the main menu Measure manual mode Select the type of measurement or press STOP to cancel measurement 91 7 2 Manual operation Measure an assay GLPtest Measure background Normalize Make a normalization tray Press START to measure Select Measure Background The following display is shown Measure background manual mode Make sure that there are no samples in the detectors and then press the START key If you want to return to the previous menu press the EXIT key To cancel the measurement press the STOP key Press the START key The display shows the main menu The text Measuring background in the first status line indicates that the background measurement is active You can do other tasks while the background is being measured If you want to see counting results select Show CPM results Wait until the complete normalization report is printed out 7 2 9 Manual GLP normalization See chapter 6 2 for the principles of GLP normalization and section 7 2 7 for details of the practice 7 2 10 Detector usage When some detectors are inactive the conveyor can only use 1 2 or 5 detectors of a 10 detector instrument This can result in that there may be detectors that are active but nevertheless cannot be used in measure
119. each label Protocol B must be defined before A otherwise you cannot select the B protocol when you edit the A protocol The explanation following applies to both the internal RiaCalc WIZ software and the external MultiCalc software Note In dual label RIA IRMA RATIO assays a warning is printed if spill over cannot be eliminated from measured data or if the elimination would cause results to be inaccurate 7 1 2 Setting dual label protocols The following example shows setting of the protocol for a dual labelled B12 Folate assay Create a protocol with the name B12 and ID 11 Answer YES to the question DUAL LABEL Two extra lines appear e g Channel A Chn B Protocol B12 Select Channel B 85 7 1 Dual label counting The chn B line will disappear Give the coding for B12 standards and controls as in a single label assay protocol Select the label First Isotope I 125 Second Isotope Co 57 Give the rest of the parameters including time and coding for the Folate standards and controls as in the case ofa single labelled protocol Exit and Save the protocol Make a copy of this protocol with the name Folate and ID 10 Answer YES to question DUAL LABEL Two more lines appear Select Channel A Chn B Protocol B12 Make other necessary changes to for example coding Note The protocol types for A and B can be different i e one can be RIA and the other IRMA Exit and Save the protocol Run the assay as for single labe
120. ector had a sufficiently high activity for this isotope so that the peak exceeded ThresholdChannelCounts If the parameter SYSTEM Operating mode Default is norm window is No the expected isotope peak positions are stored between assays that use the same isotope If there has not been any sufficiently active sample in this detector since isotope normalization the peak position during normalization is used as the expected position If the parameter Default is norm window is Yes the peak determined at isotope normalization time is taken as the expected peak position at the beginning of every assay If SYSTEM Isotope lt isotope name gt Spectrum type is I 125 and the smoothed spectrum height of the isotope main peak exceeds ThresholdChannelCounts then the existence and height of the coincidence peak is checked If the coincidence peak is not high enough the spectrum is considered bad The coincidence peak checking is done in the same way as in I 125 isotope normalization see Section 10 2 4 2 1 7 on page 140 Then if the smoothed spectrum height of the isotope main peak exceeds ThresholdChannelCounts this limit value is increased by an amount this is equal to the smoothed spectrum height of the isotope main peak times multiplied by SYSTEM Isotopes lt isotope name gt Threshold level It is assumed that the isotope activity increases background level outside the main peak area by this amount In th
121. ed for isotope decay Then the CPM values are printed if selected in the MIA protocol The title used for these values is CCPM 10 4 3 The printout fields DPM RATIO and CCPM DPM Ina MIA protocol these fields can selected to be included in the MIA printout DPM is CCPM the corrected CPM divided by the actual isotope window coverage and by the parameter SYSTEM Isotopes lt isotope name gt Efficiency The actual isotope window coverage is the ratio of counts in the isotope window to counts in the open window that contains all channels of the multi channel analyzer You can print the actual coverage during isotope normalization by setting SYSTEM Diagnostic output Print diagnostic info to YES If SYSTEM Isotopes lt isotope name gt Counting window is Dynamic then actual coverage is always slightly larger than SYSTEM Isotopes lt isotope name gt Window coverage RATIO is the relative amount of isotope present in an unknown sample given by CCPM as compared to the standard sample for this isotope The standard sample CPM is included in isotope normalization printout The ratio CCPM DPM gives the absolute efficiency of the instrument using the current isotope counting window 10 4 4 Replicate averages In a MIA protocol the number of unknown replicates can be specified If it is greater than 1 unknown replicate averages are calculated The averages are weighted according to the measurement time used to me
122. emoved and counting of the current batch resumed To initiate STAT counting during normal automatic counting press the STAT key on the built in keyboard or FS on the external keyboard Any samples in the detectors will be removed and returned to the racks The sample elevator arm will be raised as in manual counting then the display will prompt you to set three parameters These are the isotope to be used the counting time the maximum counts limit Select Do STAT and press E When you have set these parameters load your samples in the sample tray and place the tray in the detector wells Press START Samples will be counted and results will appear on the display The five display modes are described in section 2 4 2 Results will be sent to the printer First the heading STAT ASSAY will appear Then the results will be printed out The output items are DET PEAK CHN PEAK DEV RESOL ACTIVITY COUNTS EFFICIENCY ERROR where ERROR 100 Counts WINDOW keV LOW HIGH DECAYED ACTIVITY MEASURED COUNTS DETECTOR EFFICIENCY RELATIVE ERROR HORROCKS EFFICIENCY SIGNIF LEVEL these only appear if extended normalization printout has been selected these two items do not appear for single detector instruments If the instrument has several detectors installed but only one of them is active these fields are printed however Then efficiency is 1 0 and the error 0 0 At the end ofthe STAT printout the w
123. ensive 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 radioisotope 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 IAEA 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 if the minimum number of extra surfaces is introduced into the room Lastly it is extremely important to store all radioactive materials in a separate room to which access is restricted 113 7 8 Large Eppendorf tubes 7 8 Large Eppendorf tubes 7 8 4 Introduction Note this feature should be installed by a service engineer It is possible to measure large Eppendorf tubes These have caps and can be inserted only at every other sample position in the rack at odd sample positions 1 3
124. enu Show cpm results Show evaluation results Operate conveyor RiaCale WIZ Press START to measure 4 Check the time and date by selecting SYSTEM mode and DATE If it is not correct then give the correct value See System Time amp Date setting Return to the main display by pressing EXIT 5 Make sure that the counter is in the right mode i e that the text RiaCalc WIZ appears near the bottom of the screen see the figure above If it does not then select SYSTEM and then Operation mode The evaluation must be RiaCalc WIZ 4 3 Normalization Make sure that WIZARD has been normalized for the isotope you are going to use in your measurement See chapter 6 Normalization 4 4 Protocol editing 4 4 1 What is a protocol The conditions controlling samples e g counting time curve fitting method etc are stored in a protocol Before counting can be done a protocol must exist Each protocol must have a name and an ID number This number allows the protocol to be called into use by means of a barcode label To access protocol file handling you must first select the PROTOCOL option on the display 39 4 Operation with internal RiaCalc WIZ 4 4 2 Protocol operations There are several commands available to use with a protocol the most common ones are CREATE making a new protocol and EDIT change the contents of an existing protocol The complete list that appears after you select PROTOCOL is as follows
125. er items in the menu are used to select the isotope and one of the stored values mentioned above In the same menu that is used to set the limit values you can specify that a warning message a graph or both are printed if during GLP test measurement some quantity is not within limits 80 6 2 GLP test normalization 6 2 5 Viewing GLP data The stored GLP test normalization data can later be viewed graphically by setting the FILES menu item GLP data Operation to View and pressing the Enter key when the highlight bar is on the menu item Do operation The other items in the menu are used to select the isotope detector ifa 1470 instrument has more than one detector installed and one of the stored values mentioned above 6 2 6 Outputting GLP data GLP plots can later be printed by pressing the digit key 6 while the plot is displayed The plot is sent to the printer that is connected to the WIZARD printer port ifthe SYSTEM parameter Printout selections Use printer port is Yes The plot is also sent via the WIZARD PC port to MultiCalc if either SYSTEM Operation mode Evaluation is MultiCalc or SYSTEM Printout selections Without buffering to PC is Yes When the plot is sent to MultiCalc special code characters are added to the data so that the plot can be printed by the printer that is connected to the PC running MultiCalc In order for the printing to succeed MultiCalc must be receiving data from WIZARD and in the WI
126. erations 7 3 2 Date The instrument has a built in clock The clock keeps time also when the power is switched off If the power is off for an extended period of time several months it may be reset After that you must set it again Proceed as follows Select SYSTEM and DATE A display appears System date time The current date amp time is Thursday April 12 1990 1 23 pm To set new date time select the last line in the menu and press ENTER New year 1990 New month April New day 12 New hour 1 pm New minute 23 Set new date amp time Choice use lt gt or Enter To change a number select the appropriate line with the up or the down arrow key and change the value with the left or right arrow key To make time amp date selection permanent Select Set new date amp time and press E 7 3 3 Isotopes WIZARD has a built in library which consists of the optimized counting conditions for a large number of number of isotopes see chapter 9 Specifications The list of isotopes also shows if an isotope is normalized or not Remember that an isotope must be normalized before you can use it Each isotope is defined by many parameters described below In normal counting there is no need to change them because they are given optimum values in the factory If for some reason you want to make changes you can 93 7 3 System mode do so Note however that any changes will often invalidate the
127. ess the START key and select the appropriate item from the menu If an isotope or background is normalised or a GLP test measurement made while the rack id reader is not in use there is no need to add an empty rack after the measurement rack in order to stop the conveyor 7 3 8 2 Key click YES enables and NO disables the audible signal when the keyboard is used 7 3 8 3 Nominal gain This parameter lets you change the nominal gain value used by the program At the factory the value is preset to 1 00 keV per channel If this value is changed a corresponding change must be made to the instrument hardware 7 3 8 4 Counter number This parameter gives the instrument identification number in printouts This is needed when several counters are used The value is printed only if it is greater than 0 See 7 3 9 8 7 3 8 5 Use end transfer lane When this is set to YES racks are free to circulate around the conveyor including moving from the end of the output lane to the beginning of the input lane If you select this parameter to be NO the end transfer lane will be disabled and racks which have passed through the counting position will just stop in the output lane 102 7 3 System mode 7 3 8 6 Recover from power failure Recovering from power failure can be enabled or disabled by setting this parameter to YES or NO respectively 7 3 9 Diagnostic output This allows you to get a printout of diagnostic information for all detectors specified by your
128. essing is to be done but if parallel processing is done then the channel A settings dominate CODING For parallel processing CODING must be identical except for the numerical values for concentrations of standards Unknowns must begin from the same position number and must have similar structure number of replicates and dilutions with the same dilution factors 87 7 2 Manual operation 7 2 Manual operation 7 2 1 What is manual mode In manual mode the automatic sample changer is not used but the samples must be loaded into the detector wells by the operator using a sample tray The manual mode therefore only affects the way samples are moved to and from the detectors the actual counting is exactly as in the normal i e automatic mode 7 2 2 When to use manual operation You may wonder when to use manual mode since WIZARD is an instrument which offers automatic operation The following are examples of occasions when manual mode should be used in the case of conveyor contamination to count samples in large vials over 13 mm in diameter or over 90 mm in height in case of in conveyor malfunction until the service personel have cleared the problem to count extremely active samples when additional attenuation is needed 7 2 3 How to set manual mode Select SYSTEM First the Evaluation mode selection appears The three possible modes are CPM WIZARD produces CPM results These may be evaluated in another computer O
129. f all measured assays isotope normalisations and GLP TEST measurements can be printed or stored on a datalogger disk The file can also be deleted or sent via the PC port to an external PC The waste log can contain approximately 700 entries after this the older half of the entries is deleted and the log starts growing again For isotope normalization and GLP test TOTAL CPS is the average corrected sample activity in the isotope counting window in all detectors used It is corrected for dead time background activity and isotope decay TOTAL DPS is the average corrected sample activity in the open window divided by Efficiency which is a SYSTEM parameter that appears after you select Isotope and then the isotope name For assay measurement TOTAL CPS is the sum of all printed corrected CPM values of measured tubes in the assay The DPS value is obtained by dividing the printed corrected CPM by the actual coverage of the isotope counting window and by the parameter Efficiency refered to above TOTAL DPS is thus the sum of all DPS values of measured tubes in the assay 5 7 Real time clock The real time clock of the counter is set from MultiCalc through a command included in the communcation protocol 70 5 Operation with external MultiCalc If for some reason you want to set the clock manually you can do it by giving one of the following commands when MultiCalc is in terminal mode CLOCK dd mo yy hh mi ss This sets the data and time
130. f this parameter is set to YES background correction is not made for single and dual labelled RIA IRMA RATIO assays and isotope normalizations Background correction is also not made made when spillover or crosstalk correction coefficients are calculated for single and dual labelled RIA IRMA RATIO assays When you set this parameter back to NO you must redo all isotope normalizations that have been done after this parameter was set to YES so that background correction is applied to spillover and crosstalk correction coefficients Crosstalk correction is done only in 1470 multi detector models 7 3 10 Communication parameters 7 3 10 1 Three output ports At the back of the instrument there are two RS 232 serial ports and one parallel port for output devices 104 7 3 System mode PRINTER serial 1 PC 2 PRINTER parallel The PRINTER ports are to be used with IBM EPSON compatible dot matrix printers or HP DeskJet and LaserJet printers and the PC port with personal computers actually it can be used with any computer having an RS 232C interface The PC port has a simple ASCII communication and to communicate with the 1470 at least a terminal emulator program is needed in the external computer The main use of the PC port is to connect the MultiCalc to the WIZARD 7 3 10 2 Communication parameters For successful communication the communication parameters must be set to be the same at both ends In System mode communica
131. for both isotopes are printed separately e g 1 1 125 2 Co 57 Load You can load a single protocol or all protocols from an external microfloppy disk Such protocols will have either been saved there using the Save function see below or will be from MultiCalc In addition to the protocols themselves any associated data will also be loaded according to what you select as follows Normal Load will give you 1 Protocol files 2 Controls files 3 Trends files Extended Load will in addition give you 4 Standard curve files 5 Data files 6 Results files Note if you load a new protocol when a previous one with the same name or ID number exists in the instrument you have the choice of either renaming the protocol to be loaded or deleting the previous protocol Save This function allows you to transfer files from WIZARD to an external microfloppy disk for storage Subsequently these files can either be loaded back into WIZARD or into MultiCalc As with Load there are two options Save or Extended Save In the former case only protocols controls and trends will be saved In the latter case standard curves data precision profiles input files and results files will be saved also Purge Deleted protocols and associated data files can be permanently erased from the instrument hard disk with this function Password When a protocol is created you can give a two character password All characters are allowed in the password and the
132. he channels named by Resol start chn and Resol end chn in the diagnostic info printout give the interval over which the curve is fitted during resolution calculation The values a and a of a least squares regression line can be determined from the matrix equation PW Wx a AW 2x Wa H dX where x is the number or the energy of the ih channel in the interval fj In Q xj Wj i is the value of the weight function for channel i yj activity or counts in the ith channel The weight function was chosen to emphasise the importance of points that lie near the top of the peak 143 10 2 Isotope normalization and GLP TEST measurement Solving for a and a we get a o mo meo EWS ma a momo omm D ZW Wai QW The resolution values calculated with this method are fairly large for the I 125 and 1 129 isotopes Therefore the calculated resolution values are multiplied with 0 83 Resolution is not calculated if SYSTEM Isotopes isotope name gt Counting window is Fixed References Monidetektorinen gammalaskin A multi detector gamma counter by Tapio Yrj nen Pro gradu work at Turku University 1984 Pages 68 70 Evaluation of photopeaks in scintillation gamma ray spectroscopy by W Zimmermann Rev Sci Instrum 32 1063 1961 Fitting of Gaussian to peaks by non iterative method by T Mukoyama Nuclear Instruments and Methods 125 289 1975 Determination of peak area by L Kokta Nuclear Instru
133. he external keyboard to speed up accessing of items Home End Page Up and Page Down Press Exit twice Notice that the part of the original list CPM counts per minute chn A CONC concentration chn A has changed to a new list CPM counts per minute chn A ETIME elapsed time in decimal hours CONC concentration chn A 50 4 Operation with internal RiaCalc WIZ 4 6 7 Pasting a cut item The item most recently deleted with the cut item instruction can then be pasted back by selecting Paste item after this one 4 6 8 Tabulation of output fields Each printout field has a maximum field length for example time has a length of 5 characters It takes this space even if the number of digits in the actual printout is less A field is separated from the previous one by one space from the previous value You can also define the position of the output field by giving a number called a tabulation number There are two approaches to tabulation either giving an absolute position on a line in terms of the number of characters from the beginning of the line to the beginning of the field or the number of characters between the beginning of the field and the end of the previous one The former is called absolute tabulation and the latter relative tabulation The default setting is one space from the previous value Example There is the selection SEO sequence number PAT patient number TIME counting
134. he list but they cannot be stored Replicates Lets you specify a replicate value for controls 1 99 but no dilution Alternatively you can specify that the patient replicate and dilution from the coding part be used 4 5 11 Saved files You can specify which types of files are required These protocols are generated automatically during evaluation The choices are Input data The data used for analysis See Ria evaluation from file Standard curve Standard curve Trend data Results for trend profiles Control samples data Concentration results of control samples Results data Results for output to an external computer This file is an ASCII file and its contents are selected on the line Result file The default for each file type is NO not selected 4 5 12 Display This parameter line allows you to specify which results are to be displayed on the screen 4 5 13 Printer This allows you to specifiy the results for printout The list of items which from which selection can be made are the same as the list for the display but the actual selection for the printer may be different from that for the display 4 5 14 Results files This allows you to specify the contents of the Results ASCII file The possibilities are the same as those for the display but the actual selection for the results files may be different from that for the display See the section entitled Output editing for a description of the types of output options ava
135. he sum of all printed corrected CPM values of measured tubes in the assay converted to CPS The DPS value is obtained by dividing the printed corrected CPM converted to CPS by the actual coverage of the isotope counting window and by the parameter Efficiency referred to above TOTAL DPS is thus the sum of all DPS values of measured tubes in the assay 58 4 Operation with internal RiaCalc WIZ 4 10 Selectable outputs TO Move to this column LEFT Move to the left RIGHT Move to the right LINE Move the rest of the text to a new line t Vectors Print sample output result items e g POS TIME vertically K 1 Rack number 3 digits for both groups no average 1 Detector number for both groups no average 111 Sequence or tube number 0 111 Patient number with both averages unknowns only E RS 1 Counting time in seconds ETIME 1 Elapsed time in decimal hours COUNT 11 Total counts and first average COUNT B 11 Total counts and first average on B channel CPM 11 Corrected CPM and first average B 11 Corrected CPM and first average on B channel 111 Concentration with both averages B 111 As above for channel B 011 Coefficient of variation as a percentage only averages B 011 As above for channel B E 0 01 CV from reference precision profile average for only unknowns E B 0 01 As above for channel B E 111 Code text from coding and control list
136. hese values are stored in the file WASTELOG TXT in the root directory of the instrument hard disk TOTAL CPS is the average corrected sample activity in the isotope counting window in all detectors used It is corrected for dead time background activity and isotope decay TOTAL DPS is the average corrected sample activity in the open window divided by SYSTEM Isotope lt isotope name gt Efficiency 10 2 15Storing GLP TEST normalization results Instrument performance can be monitored by running GLP TEST normalizations at regular intervals These store data that can later be viewed in graphical format GLP means Good Laboratory Practice A GLP TEST normalization is similar to isotope normalization only results are stored differently Data obtained in a GLP TEST normalization is not used in assay measurements but is stored so that it can later be compared with other GLP TEST normalizations using the same isotope This comparison is done by presenting the values of some measured parameters as a function of time so that any systematic trends or large random deviations can easily be discerned A GLP TEST measurement rack has only one holder and sample at the last position of the rack The rack has a clip with the TEST instruction at the RACK SPECIAL position and the isotope code at the PROTOCOL position All isotopes that can be normalized can also be used for GLP TEST measurements The counting time is set by the parameter SYSTEM
137. his in turn is used when counting windows are converted between keV s and channels e Horrocks efficiency is determined if SYSTEM Isotope lt isotope name gt Spectrum type is I 125 and Counting window is Dynamic or Dynamic keV This makes it possible to determine the absolute detector efficiency for I 125 without using an external standard e If Counting window is Dynamic the counting window in keV s is determined for each detector This is done so that starting from the isotope main peak the window is widened until it includes at least the fraction given by SYSTEM Isotope lt isotope name gt Window coverage of all counts in the spectrum If Spectrum type is I 125 both the main peak and the coincidence peak are always included in this counting window e Relative detector efficiencies are determined for 1470 counters that have multiple detectors installed e Measured isotope normalization sample spectra are stored for all detectors This makes it possible to eliminate spillover in dual label RIA IRMA RATIO assays and 1480 multiple isotope assays MIA If a 1470 counter has more than one detector installed and SYSTEM Isotope isotope name gt Crosstalk correction is Yes then for each detector where the normalization sample 1s measured the spectra of all other empty detectors are also stored in the instrument hard disk These are used to eliminate crosstalk in single and dual label assay counting
138. his is the fraction of counts in the whole spectrum that falls in the isotope counting window Background activity is ignored when this is calculated e Detector stability probability This can be calculated if the parameter SYSTEM Isotope isotope name gt Repeat times is greater than 1 The measured counts in repeat measurements are compared with each other and the program calculates the probability that a Poisson distributed random variable having the same mean would have had a greater variance than was observed in these repeat measurements This probability is called Significance level and its unit is If it is often near zero or near 100 this means that there is systematic error in repeat measurements The number stored is transformed from the Chi square probability that is shown in the printout SIGNIF LEVEL so that 5 corresponds to 50 4 to 10 3 to 1 2 to 0 1 1 to 0 01 0 to lt 0 001 6 to 90 7 to 99 8 to 99 9 9 to 99 99 and 10 to gt 99 999 This is to make very small and large probability values stand out more clearly e Measured CPM in counting window This is corrected for dead time and background activity but not for relative detector efficiency or isotope decay e Measured total CPM in the whole spectrum This is equal to measured CPM in the counting window which has been corrected for isotope decay and divided by window coverage For each of the above quantities one can set a low and a high limit by
139. ht end of the CPM printout and as the last item in datalogger files Set the parameter to YES to enable the field and to NO to disable it Measurement start time can also be sent to MultiCalc if the mode is MultiCalc but the field can be enabled and disabled only from MultiCalc the parameter Print meas start time has no effect in this case See chapter 5 Operation with MultiCalc for more details 7 3 9 7 Instrument serial number This parameter shows the instrument serial number that has been stored in read only memory You cannot edit this number The instrument serial number is included when the instrument model and program number are printed See the next parameter for details 103 7 3 System mode 7 3 9 8 Print model and version The counter model program version and instrument serial number are printed before normalizations and assay measurements The printing can be disabled by setting this parameter to No The string has following form 1470 10 detectors RiaCalc WIZ program 3 4 serial 667273 counter 2 If the counter has only one detector installed the number of detectors is not mentioned RiaCalc WIZ means that RiaCalc WIZ is enabled The Serial number is present only if it has been stored in the counter s read only memory The counter number is printed only if it is gt 0 If not disabled this string is printed before background normalisation isotope normalisation GLP TEST normalisation and RIA IRMA RATIO a
140. icate sample CPM s were obtained using the same counting time But implicit in this is the assumption that the background CPM s that were subtracted from individual replicate sample CPM s were all measured separately This would be equivalent to the total background counting time for the replicate average being obtained from the number of replicate tubes multiplied by the background counting time used for a single replicate tube This would have greater accuracy but in fact we have only measured background once The formulae above takes this into account 176 10 5 Appendix Some basic mathematical formulas 10 5Appendix Some basic mathematical formulas 10 5 1 Error calculation Isotope decay has Poisson statistics so that the variance of a measured counts value is equal to the counts value itself Standard deviation is by definition equal to the square root of variance Relative error is defined to be equal to standard deviation divided by the counts value If a random variable is multiplied by a constant the corresponding variance is multiplied by the square of that constant Iftwo independent random variables are summed their variances are also summed to get the variance of the sum value 10 5 2 Matrix calculation formulas The product of an NXN matrix with an 1xN vector is defined as follows Yo m My Vi ee My Muy Vy S my i We can thus write a system of N linear equations each having N
141. ich may appear during editing are Make new line after this one You can insert a new line after the currently selected one by invoking this menu item Chn A cpm Enter here the CPM value for channel A If CPM fields for both channel A and channel B are empty it means that the measured tube was missing Chn B cpm Enter here the CPM value for channel B if you want to use it Cut this line If you invoke this menu item the line selected for editing is deleted to a cut buffer It can be later pasted by choosing the menu item Paste item after this one Paste line after this one If you invoke this menu item the previously deleted line is pasted from the cut buffer after the line that was selected for editing Jump to a line If you invoke this menu item you are asked to give the number of the line to where you want to jump Then you must press the EXIT key When you have finished editing press the EXIT key In the case of the Copy operation the file operations submenu includes the following additional lines Copy to protocol Specifies to which protocol the copy of the file is to be attached Copy to run id Specifies the run id number that the copy of the file is to have If the protocol is the same as for the copied file then the run id must be different When you access the list of possible run ids then you will see that used run ids do not appear on the list so you cannot duplicate them When data has been saved in an input file 1t can
142. iency by the detector and the isotope counting window used Detector efficiency depends on the counting window and may also change with time We assume that detector efficiencies have not changed between the two isotope normalizations The following 2 N x 2 N matrix of CPM activities in counting windows is set up where N is the number of detectors that are in use Isotope A normalization sample is in detector 1 Spectrum is taken from detector 1 Counting window is for isotope A Isotope A normalization sample is in detector 1 Spectrum is taken from the last used detector Counting window is for isotope A Isotope A normalization sample is in detector 1 Spectrum is taken from detector 1 Counting window is for isotope B Isotope A normalization sample is in detector 1 Spectrum is taken from the last used detector Counting window is for isotope B Isotope A normalization sample is in the last used detector Spectrum is taken from detector 1 Counting window is for isotope A Isotope A normalization sample is in the last used detector Spectrum is taken from the last used detector Counting window is for isotope A Isotope A normalization sample is in the last used detector Spectrum is taken from detector 1 Counting window is for isotope B Isotope A normalization sample is in the last used detector Spectrum is taken from the last used detector Counting window is for isotope B Isotope B normalizat
143. ies too so the system can also include measuring devices for almost all other label technologies 8 6 Customization and upgradability A number of versions of the instrument are available with 1 2 5 or 10 detectors offering throughputs from 50 to 500 samples hour The smaller conveyor size can take 550 samples when fully loaded The larger model can take 1000 samples This combination of operating systems and physical configurations offers you a wide range of possibilities to choose from You can select the one which best suits your current needs and resources knowing that when necessary you can upgrade the system to meet new requirements 8 7 Compactness WIZARD has an exceptionally small footprint for a multi detector automatic gamma counter the 550 sample 10 detector table top version of WIZARD is only 65 cm wide by 70 cm deep This has been achieved by locating the 119 8 Instrument description display and detector block between the lanes of the conveyor rather than adding them onto the end thus increasing the instrument length 8 8 High efficiency detectors In the past automatic multi detector counters have been based on the incomplete geometry of through hole detectors but this is not so in the case of WIZARD PerkinElmer Life Sciences Wallac Oy has successfully incorporated well type detectors into the design of a high throughput automatic gamma counter These give WIZARD the best possible counting geometry and hence high
144. ilable and how to select and edit them 4 5 15 Coding The program must know in which order RIA standards are loaded and what their nominal concentrations are This information is supplied in the CODING Other special samples such as Blank and Reference can also be specified 45 4 Operation with internal RiaCalc WIZ When you are creating a new protocol there is a default coding available as follows lt Start of assay gt 2 BLANK REFER TOTAL STD1 STD2 STD3 STD4 STD5 16 STD6 STD7 UNKN 1 ll ll o NA ll Ww No NO NO NO ho Po PO PO PO DN PO ll md gt The number on the left is the replicate number and the number in the right is the nominal concentration The replicate number preceding UNKN1 means that all unknowns have this number of replicates In the example above all samples are duplicates To change a replicate number use the keys to increase or decrease respectively the number of replicates Use the numerical keys to change concentration values Use the delete key to remove a value If you want to edit these parameters or any other already created set of parameters use the cursor control keys to select the item to be edited and press Enter This will cause a display like the following to appear Edit protocol 11 PROTO3 Edit item Select what to do with the item that was selected in the coding list New replicate No 02 New item t
145. imit counts value that the smoothed spectrum height of the isotope main peak must exceed This is done with the following formula ThresholdChannelCounts SignifCpm NominalGainForTheUsedEnergyRange CountingTimelnSeconds 60 where SignifCpm SYSTEM Isotopes isotope Signif cpm per keV NominalGainForTheUsedEnergyRange is explained in Section 10 1 4 on page 132 If the smoothed spectrum height of the isotope main peak does not exceed ThresholdChannelCounts the whole spectrum is checked for peaks that exceed ThresholdChannelCounts If such a peak is found the spectrum is classified as bad otherwise it is considered to have too low activity In both cases the isotope normalization is aborted Sometimes a spectrum may be classified as bad although it in fact is just too weak This happens if SYSTEM Isotopes lt isotope name gt Spectrum type is Many peaks and there are peaks outside the interval given by SYSTEM Isotope lt isotope name gt Peak pos keV and SYSTEM Isotope lt isotope name gt Max norm dev that are higher than the isotope main peak If SYSTEM Isotopes isotope Signif cpm per keV is so large that the height of the isotope main peak is smaller than ThresholdChannelCounts but some other peak in the spectrum is higher than this then the spectrum is classified as bad although actually it is too weak In this case you have to decrease Signif cpm per keV so that the mai
146. in the instrument hard disk Latest background spectra are also copied and stored with normalization spectra These are used to calculate spillover correction coefficients at the beginning of dual label RIA IRMA RATIO assays and 1480 multiple isotope assays after it has been determined what fraction of one isotope s counts falls into another isotope s counting window 10 2 13Storing crosstalk spectra to enable crosstalk correction If in the 1470 counter model SYSTEM Isotopes lt isotope name gt Crosstalk correction is Yes then every time a normalization sample is measured in a detector spectra from all used detectors in the counter are stored This makes it possible to determine how much crosstalk there is between detectors The crosstalk correction factors for single label RIA IRMA RATIO assays are calculated during isotope normalization For dual label RIA IRMA RATIO assays the correction factors are calculated at the same time as spillover correction coefficients that is the first time such an assay is measured after one of the isotopes has been normalized The single label crosstalk correction factors are calculated in the following way The program has already determined the counting windows in terms of MCA channels and the relative detector efficiencies for all detectors To determine the crosstalk from detector K to detector L the spectrum measured in detector L when the normalization sample is in detector K is looked up Counts are s
147. indow types take into account that the effective gain is not always exactly the same as the nominal gain but may vary according to e g temperature and measured activity The effective gain is calculated by dividing the main peak energy in keV s by its observed channel number To do this the program must know the peak energy and be able to find the actual MCA channel number of the peak The peak energy is given by the parameter Peak pos keV The next sections explain how the peak channel number is determined Note If SYSTEM Operation mode No dynamic normalization is Yes then the same channel number will be used as was found during isotope normalization This means that counting windows remain the same as they were during isotope normalization they are not shifted anymore during assay measurement 10 3 2 2 1 Single label assay The isotope main peak channel number is determined in the same way as in isotope normalization see Section 10 2 4 2 1 on page 138 Only the parameter Max norm dev in Section 10 2 4 2 1 3 on page 139 is replaced with the parameter Max assay dev If the smoothed spectrum height of the isotope main peak does not exceed ThresholdChannelCounts see Section 10 2 4 2 1 6 on page 140 the activity of the sample is considered to be so low that the so called expected peak position is taken as the position of the peak The expected peak position is taken from the latest batch where a sample in this det
148. ion sample is in detector 1 Spectrum is taken from detector 1 Counting window is for isotope A Isotope B normalization sample is in detector 1 Spectrum is taken from the last used detector Counting window is for isotope A Isotope B normalization sample is in detector 1 Spectrum is taken from detector 1 Counting window is for isotope B Isotope B normalization sample is in detector 1 Spectrum is taken from the last used detector Counting window is for isotope B Isotope B normalization sample is in the last used detector Spectrum is taken from detector 1 Counting window is for isotope A Isotope B normalization sample is in the last used detector Spectrum is taken from the last used detector Counting window is for isotope A Isotope B normalization sample is in the last used detector Spectrum is taken from detector 1 Counting window is for isotope B Isotope B normalization sample is in the last used detector Spectrum is taken from the last used detector Counting window is for isotope B Only detectors that are in use appear in the matrix If for one of the isotopes say B Crosstalk correction is No then all elements which have the text Isotope B normalization sample is in detector X Spectrum is taken from detector Y with X different from Y are equal to 0 Then values in each column of the matrix are divided by the diagonal value in that column This converts measured activities to
149. ioned in the diagnostic info printout 139 10 2 Isotope normalization and GLP TEST measurement 10 2 4 2 1 5Finding the I 125 coincidence peak If SYSTEM Isotopes lt isotope name gt Spectrum type is I 125 the program determines the position and height of the coincidence peak The expected position of the coincidence peak is twice the channel number of the isotope main peak This should be twice the keV value but detector unlinearity is not taken into account here Both the main and the coincidence peak actually consist of two peaks that are near each other but since the detector cannot resolve them from each other the program can ignore this The interval where the coincidence peak is searched is IntervalLowBorder ExpectedCoincChannel 1 0 MaxCoincDev 100 0 IntervalHighBorder ExpectedCoincChannel 1 0 MaxCoincDev 100 0 where MaxCoincDev SYSTEM Isotopes isotope Max coinc dev Of all the channels in the interval that are marked as Maximum the one with most counts is taken as the coincidence peak If no such channel exists in the interval the spectrum is considered not to have a coincidence peak and in this case the normalization is aborted Reference Monidetektorinen gammalaskin A multi detector gamma counter by Tapio Yrj nen Pro gradu work at Turku University 1984 Page 72 10 2 4 2 1 6Checking whether the isotope main peak is high enough First the program calculates a l
150. is used as the effective gain Let us call the result ExpectedPeakChannel Then the interval boundaries are calculated with the following formulas IntervalLowBorder ExpectedPeakChannel 1 0 Max norm dev 100 0 IntervalHighBorder ExpectedPeakChannel 1 0 Max norm dev 100 0 10 2 4 2 1 4Finding the peak in the expected interval Of all the channels in the interval that have previously been marked as Maximum the one with most counts in the smoothed spectrum is taken to be as the isotope main peak If no channels marked as Maximum exist in the interval the whole spectrum is checked for peaks that exceed ThresholdChannelCounts see Section 10 2 4 2 1 6 on page 140 If such a peak is found the spectrum is classified as bad otherwise it is considered to have too low activity In both cases the isotope normalization is aborted But if the isotope main peak is found the program seeks the nearest channels to the left and right of the peak channel that has been marked either as Maximum or Minimum If either one is a Maximum the peak is considered abnormal Otherwise they are considered to be the start and end point of the peak They are named in diagnostic info printout as Peak begin channel and Peak end channel If there are between Peak begin channel and Peak end channel channels that have been marked either as Small min or Small max then the peak is considered to have noise This is also ment
151. is the older half of the entries is deleted and the log starts growing again For isotope normalization and GLP test TOTAL CPS is the average corrected sample activity in the isotope counting window in all detectors used It is corrected for dead time background activity and isotope decay TOTAL DPS is the average corrected sample activity in the open window divided by Efficiency which is a SYSTEM parameter that appears after you select Isotope and then the isotope name For assay measurement TOTAL CPS is the sum of all printed corrected CPM values of measured tubes in the assay converted to CPS The DPS value is obtained by dividing the printed corrected CPM converted to CPS by the actual coverage of the isotope counting window and by the parameter Efficiency refered to above TOTAL DPS is thus the sum of all DPS values of measured tubes in the assay 35 4 Operation with internal RiaCalc WIZ 4 Operation with internal RiaCalc WIZ 4 Operation with internal RiaCalc WIZ 4 1 Introduction The instructions here describe the routine operation of WIZARD when it is running the internal RiaCalc WIZ software 4 2 Start up 1 Switch on the printer this should have already been connected to the counter 2 You can put the data disk named 1470 Datadisk into the WIZARD disk drive The name label should be facing upwards 3 Switch on WIZARD After about 3 min the display will show 1470 Main Menu QA PROTOCOL FILES SYSTEM Subm
152. isplay will revert to the Edit Print Undo Exit options You must use this option to exit from Editing 4 9 6 Control files These are produced as part of the quality control operations of RiaCalc WIZ They can be viewed or deleted with this file function Control type This allows you to specify the control type that is to be operated on In principle there are six types LOW MEDIUM HIGH CONTROLA 5 and 6 However the actual types which appear for viewing are only those which were selected in protocol setting 56 4 Operation with internal RiaCalc WIZ When you have selected the type and do operation the control plot will appear allowing you a number of functions for editing the plot These functions are the same as for trend plots and are described below 4 9 7 Trend files Trend files can also be viewed or deleted after they have been produced Trend type Specifies the trend type that is to be operated on This list is fixed because unlike controls it is not affected by the protocol settings The options available are Slope at ed 50 Y intercept Estimated concentration at 20 binding Estimated concentration at 50 binding Estimated concentration at 80 binding Blank over Total ratio Reference over Total ratio Blank over Reference ratio Minimum detectable concentration Parallelism factor Difference slope Histogram 4 9 8 Control and trend plot editing The functions available for editing control and tren
153. ithout a holder are omitted in position counting If Ignore even sample pos is YES then only even sample positions are omitted in position counting and the rack is also checked so that there are no holders in even rack positions 7 8 3 Optional detector removal Since now only detectors at odd detector well positions can be used the detectors at even well positions can be removed although they need not be If they are removed then set the parameter SYSTEM Active hardware Dets Dets only at odd positions gt only at odd positions to YES This parameter is visible only if the number of detectors installed is 2 or 5 If the installed detectors are only at odd well positions they can all be used otherwise only those at odd well positions can be used In the latter case detector numbering in assay printouts ignores detectors at even well positions If the value of either the parameter Forks only at odd positions or the parameter Dets only at odd positions is changed all normalizations will be deleted A warning is issued before this is done 115 3 Instrument description 117 8 Instrument description 8 Instrument description 8 1 Introduction In this chapter the main features of 1470 Wallac WIZARD are presented showing what they are and what benefits they offer you as a user 8 2 Self contained WIZARD has the possibility to be a completely self contained counter with its own built in computer This is a 16 bit computer h
154. l The protocol ID which in the example is 10 can be from 1 99 7 1 3 Parallel and successive evaluation The actual evaluation may take place in two different forms parallel and successive Parallel processing is when results from channel A and B are calculated simultaneously in real time Successive processing is when results from channel A are calculated and saved in real time These results are then retrieved and the results for channel B are calculated as in later Parallel processing imposes strong constraints on the assay Both analytes must have the same number of standards and controls and the number of replicates must be the same Successive processing has practically no constraints at all The number of standards controls replicate etc is freely selectable The program selects according to the protocol whether the evaluation is parallel or successive 7 1 4 Constraints on protocol setting The following is a summary of protocol commands and how these are constrained by the requirements for dual label protocols DUAL LABEL COUNTING TIME LABELS Only the channel A settings are valid The channel B settings have no effect X AXIS Y AXIS FITTING ALGORITHM STD OUTLIER REJECT CURVE EDIT HALT May be set differently for channel A and channel B protocols CONTROLS Must be set identically for both protocols if parallel processing is to occur 86 7 1 Dual label counting PRINTER Can be set differently if successive proc
155. l 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 gamma counting so you must select GAMMA Counter is the type of counter MultiCalc is to be working with You must select WIZARD Note that you can select more than one type oftechnology and counter at the same time If you have a RiaCalc or MultiCalc version that does not include a communication protocol for WIZARD you can copy it from the instrument program diskette The communication protocols for the particular program are stored on the WIZARD Main Program diskette in file 1470 C00 for RiaCalcV2 65 and file INST OCOM GAMMA WIZARD C00 for MultiCalc V2 0 186 11 Installation Write protect your WIZARD Main Program diskette and put it in drive A or B in the PC where you have RiaCalc or MultiCalc Copy the particular communication protocol to CAWIACALCIOCOM C RIACALC2 0COM if RiaCalc If MultiCalc is already installed in the PC but not for WIZARD you must go through a similar procedure to that described above but select Setup only rather than Installation from the main installation menu Start MultiCalc Press F1 COUNTER to get to counter control Move the cursor to WIZARD in the counter list Press F5 2 INSTALL to install WIZARD for MultiCalc In step a installation you were adding WIZARD to the list of possible counters to be operated with MultiCalc Now with this
156. label BKG onto an ID clip in the area marked RACK SPECIAL Fix the clip onto an otherwise empty rack Load the rack on the conveyor and press START A complete background normalization report will be printed out and background values for the entire energy spectrum will be saved in the instrument memory Since the whole spectrum is saved there is no need to make background measurements for each individual isotope 77 6 1 Normalization 6 1 3 Normalization for isotopes used Information about the isotopes to be normalized name ID number energy window settings etc must be given to the instrument before the normalization starts This information is factory set for the isotopes in the list in chapter 9 Specifications See chapter 7 3 System mode for information about isotope parameters and how to change them should you need to do so Be careful about changing isotope parameters because this will in many cases mean that you have to make a fresh normalization Remove the holders from positions 1 to 9 in a rack Place an isotope source in the last position of an otherwise empty rack The isotope should have an activity of between 50 000 DPM and 200 000 DPM Stick an ID label NORM to the ID clip in the position marked RACK SPECIAL Stick an ID label with the appropriate isotope code number to the ID clip in the position marked PROTOCOL The isotope codes are the numbers given in the table above Fix the clip to the rack load i
157. laced by the one from the default list and the display will revert to the SYSTEM function menu Note When this parameter is selected for the first time after program installation it takes a couple of minutes before the menu appears This is because the program builds an index of the isotopes in the default list To do this it must scan each protocol file and extract the isotope name and comment Since there are over 50 isotopes this takes some time After the index has been built the program writes it on the instrument hard disk When the Copy isotope menu is entered subsequently the program merely reads the index from the disk and therefore there is no apparent delay 7 3 3 10 Print isotope and normalization Note this function can be found at the end of the list of isotopes To reach it quickly press End or PgDn You can print out a list of isotopes the data from all the isotopes only normalized isotopes or one selected isotope You can choose to print the isotope parameters the measurement data or both 7 3 4 Operation mode 7 3 4 1 Evaluation mode The three possible choices are CPM WIZARD produces CPM results These may be evaluated in another computer Operation with this mode 1s described in Part 3 of this manual RiaCalc WIZ Data evaluation is carried out in WIZARD with the RiaCalc WIZ software See Part 4 MultiCalc Data evaluation is carried out in the PC using MultiCalc immunodiagnostic data management software See P
158. lect the operation and file The assay spectra text file has the following format Entities in angle brackets are written as numbers in the file Omitted lines are indicated with R lt Number of spectra in the file gt lt Number of channels in each spectrum gt 0 00 0 00 0 00 0 00 SP lt Number of first spectrum is equal to 1 gt lt Counting time in seconds gt lt Counts in first channel gt Note When assay spectra are saved in text format you can specify that all data belonging to the same spectrum is written in one line In this case the channel counts values are separated by tabulator characters This makes it easier to read several spectra at the same time into a spreadsheet program 7 3 4 4 SYSTEM Operation mode Default is norm window Ifa RIA IRMA RATIO assay has a sample with so small activity for some isotope that the main peak position cannot be determined the previous possibly shifted counting window for this isotope and detector is used If in the current assay there has not yet been for this isotope and detector a sufficiently active sample so that the main peak position could be determined then if this parameter is No the same counting window used in the most recent assay run using this isotope is used However ifthe parameter is Yes the counting window determined in the latest isotope normalization is used 7 3 4 5 SYSTEM Operation mode No dynamic normalization If this parameter is
159. licate number Elapsed time Counting time Dead time ISOTOPE COUNTS I 125 15385 Co 57 43628 Cr 51 459499 I 129 15457 Sequence number Sample number Replicate number Elapsed time Counting time Dead time ISOTOPE COUNTS I 125 242786 Co 57 425 Cr 51 955 I 129 244239 0 002 1 1 1 0 00 hours 600 seconds 0 952 BGRD CPM CPM 9 4 42132 0 38 9 128 8 88 3 0 5 9 4 42278 3 2 2 1 0 17 hours 600 seconds 1 852 BGRD CPM CPM 9 4 366 4 38 9 165119 8 88 3 989 6 9 4 367 3 3 3 1 0 34 hours 600 seconds 0 609 BGRD CPM CPM 9 4 1538 5 38 9 4350 5 88 3 46141 5 9 4 1545 7 4 4 1 0 51 hours 600 seconds 0 409 BGRD CPM CPM 9 4 24368 5 38 9 3 7 88 3 7 6 9 4 24514 3 ECPM 1 71 0 08 1 71 ECPM 76 65 56 20 0 20 CCPM 42117 9 0 5 14 1 CCPM 165123 9 212 7 28 8 DPM RATIO CCPM DPM 109101 5 1 0001 0 3860 0 6 0 0000 0 8528 80 6 0 0001 0 0565 34 0 0 0006 0 4137 DPM RATIO CCPM DPM 49 7 0 0005 0 3860 193615 5 0 9998 0 8528 224 2 0 0003 0 0565 21 2 0 0004 0 4137 DPM RATIO CCPM DPM 41 0 0 0004 0 3860 51 9 0 0003 0 8528 817572 5 0 9985 0 0565 25 7 0 0004 0 4137 DPM RATIO CCPM DPM 1099 6 0 0101 0 3860 3 1 0 0000 0 8528 18 7 0 0000 0 0565 60279 4 1 0152 0 4137 END OF MULTI ISOTOPE ASSAY Figure 13 A MIA printout The parameter SYSTEM Isotopes lt isotope name gt Counting window is Dynamic keV for 1 125 and 1 129 and Dynamic for Co 57 and Cr 51 Since 1 125
160. lization sample is in the last used detector CPM in the last used detector using the counting window of the last used detector when the normalization sample is in the last detector If some detectors are not in use the corresponding rows and columns are missing in the matrix Then values in each column are divided by the diagonal value in that column This converts measured activities to relative crosstalk values Let us denote this new matrix with the letter M If this matrix is multiplied with the following 1xN vector B Sample activity in detector 1 B Sample activity in detector 2 Sample activity in the last used detector the result is the following 1xN vector A Measured activity in detector 1 A Measured activity in detector 2 Measured activity in the last used detector 148 10 2 Isotope normalization and GLP TEST measurement Thus A MB Then the matrix M is inverted Let us denote the inverted matrix with M When this is multiplied during assay counting with a vector consisting of the measured CPM values in all used detectors the result vector consists of crosstalk corrected sample activities in those detectors that is B M A If SYSTEM Isotopes lt isotope name gt Repeat times is greater than 1 the repeat spectra are not stored after isotope normalization only the sum spectra are After a GLP TEST normalization all temporarily stored spectra are deleted 10 2 14 TOTAL CPS and TOTAL DPS T
161. lour is being measured the other assay light colour is waiting Four racks have already been counted and the fifth position in the fifth rack sample number 45 in the upper part of the figure is being counted Suppose that the power failure occurs at this moment After power is restored the sample is taken from the detector and the conveyor is rotated anti clockwise the normal direction in counting until the ID label of the first assay is encountered This assay is then counted starting with sample 1 The previously obtained 44 results can be discarded 109 7 5 Power failure Note Do not remove any racks from the conveyor before the assay to wich they belong is completed because if a power failure occurs wrong results would be obtained during power failure recovery Counting position Conveyor during power un ee failure 44 samples in assay already counted EXXIIIIXIXIII 6600606060000 sample 45 being counted 06006000000 0000000000 KIXIITIEITI 000000000991 ID2 KEXXIIIXXIIJ 0000000000 OQOOOOOO0O000 80500000000 90000000000 0000060000 O0000000009 S000000000 Counting position Conveyor after power failure Sample 1 in assay 1 being recounted 1D1 SODOGODODEO 8 0008 0088 0000000000 KXIIIIIIIII 900000090000 KXIIIIIXIIJ 9009000900009 0000 0 Cee Some 8 0008 0 C08 0000000000 9000000000 6000000000
162. ly if SYSTEM Printout selections Extended norm printout is Yes and SYSTEM Isotope lt isotope name gt Counting window is not Fixed Main peak at nominal gain is the Main peak above converted to an MCA channel number assuming that effective gain is equal to nominal gain If effective gain is different from nominal gain then PEAK CHN below is different from Main peak at nominal gain This item is printed only if SYSTEM Printout selections Extended norm printout is Yes and SYSTEM Isotope lt isotope name gt Counting window is not Fixed Nominal window coverage is the value of SYSTEM Isotope isotope name gt Window coverage It is printed only if SYSTEM Printout selections Extended norm printout is Yes and SYSTEM Isotope lt isotope name gt Counting window is Dynamic Detectors not in use lists those detectors that are not used for measurement because some detectors have been set to be inactive If normalization is done in 1470 using the conveyor there may be detectors that are active but nevertheless are not used for measurement since with the conveyor only active sets of adjacent 10 5 2 or 1 detectors can be used for measurement PEAK CHN is the observed peak channel number It is determined from the smoothed spectrum the counts values of channels on both sides of the peak channel are used to interpolate the peak position to a fraction of channel The pe
163. malization and GLP TEST measurement 0 y A e 20 027 where x is the energy and y is the activity or the number of counts in the measured spectrum From this we can derive for a general Gaussian curve 2 x xg y x 1 Q x e o y x 1 from which follows 2 f x In Q x I2 X0 This is represented by a straight line The half height width of a Gaussian curve is Hhw 20421n2 If In O x is written in the form f x In Q x aix ag we can express the resolution of the Gaussian curve in terms of a and a as follows Resolution 109 Ew 40041n2 Ja Xo do To determine a and a from a measured spectrum a straight regression line is fitted through the points x f x where the index 7 is used to number the channels in the interval over which the fitting is done x is the energy at channel i The correspondence between energy and channel number is not entirely linear see Section 10 2 4 1 2 on page 137 Next we must decide what the interval is It is taken to be the one where the smoothed spectrum channel count values are at least 67 of the smoothed spectrum height of the isotope main peak In any case the interval does not extend in either direction beyond Peak begin channel and Peak end channel If it would the peak is marked as abnormal and resolution is set to 0 If a peak is abnormal this is mentioned in the diagnostic info printout but isotope normalization is not aborted because of this T
164. matrix can be used to solve the individual Isotope activities in the sample Single labelled samples are measured in a counter having several say N detectors There is crosstalk between the detectors When the same sample is measured successively in each detector the remaining detectors being empty the crosstalk coefficients giving the relative activity measured in the empty detectors as compared to the activity measured in the detector having the sample can be determined and an NXN matrix can be set up as described in Section 10 2 12 on page 147 The crosstalk coefficients can be assumed to independent of isotope activity at least as long as the isotope activities are not very large When samples are measured in all the detectors simultaneously the inverse of the crosstalk coefficient matrix can be used to solve the activity of each of samples just as if all the other detectors were empty Samples consist of several say M isotopes and as many counting windows are used when they are measured in a counter having several say N detectors Each isotope 1s assigned one counting window There is spillover between isotope counting windows and crosstalk between the detectors When pure isotope samples are measured successively in each detector the remaining detectors being empty the coefficients giving the relative activities in all counting windows of all detectors can be determined and an M N X M N matrix can be set up as described in Section 10
165. ment 3 5 Parameters available 3 5 1 Counting time Give the counting time in seconds All samples are counted for this time The default value is 60 and the maximum value is 65000 seconds 3 5 2 Max counts limit If you want to terminate counting on the basis of the number of counts accumulated enter the counts value for this parameter Provided this number of counts is reached in each detector before the counting time expires this parameter will terminate counting If however the counting time expires first it will terminate counting even though the max counts limit has not been reached The maximum and default value for this parameter is 99999999 3 5 3 Labels If the isotope you are using is I 125 just press the down arrow key If you use another isotope change the isotope with the left arrow key or by pressing E The isotope list is shown in chapter 9 Specifications To change the contents of this isotope list go to the SYSTEM mode and the Isotopes parameter see charpter 7 3 in this manual If you want to do dual label counting select the second isotope with the right arrow key and press E or press E to get the label selection menu 3 6 Output 3 6 1 Default output In CPM mode there is a default format for output and this cannot be changed The items displayed and printed are Sample position rack number detector number batch number counting time counts CPM and CPM error In addition there are the following 3 6 2 Star
166. ments and Methods 112 245 1973 10 2 5 Summing counts over the counting window If SYSTEM Isotopes isotope name gt Counting window is Dynamic the window is determined first in terms of MCA channels and then converted to keV s During assay counting the boundary keV s are converted back to MCA channels using the effective gain at the assay counting time However if Counting window is Dynamic keV or Fixed the window is given in keV s and then converted to MCA channels The converted MCA channel values are not necessarily integer values When counts are summed over a window the boundaries of which are non integer channel numbers only a corresponding fraction of counts in the boundary MCA channels is taken to the sum This ensures that slight changes in the position or width of the counting window change the counts sum only a little 10 2 6 Background CPM in the counting window Background CPM in the counting window is obtained by summing counts in the background spectrum over the counting window and correcting the result for dead time and converting it to CPM The background dead time factor is determined when background is measured and it is stored with the background spectrum as is the background measurement time used The 1480 counter model has separate background spectra stored for both energy ranges 10 2 7 MIA standard CPM MIA standard CPM is obtained by summing counts in the measured spectrum over the isotope counting
167. ments occured just because of statistical variation This probability is called significance level and its unit is If it is near zero or near one then it means there is a systematic error in the repeat measurements See chapter 6 2 GLP normalization for more details 7 3 3 5 Crosstalk correction If you want crosstalk correction to be done when you count samples labelled with that isotope then select Yes for this parameter Yes is the default setting for isotopes with energy greater than 200 keV If you select No then no crosstalk correction will be done 7 3 3 6 Decay correction If you select this then give the isotope half life value if needed The possible values are None no decay correction is made Start results are corrected to the start of the normalization or assay Explicit the time to which the results are corrected is explicitly given If the choice Explicit is selected this keyword is not shown in the isotope editor menu but instead the time itself is shown The format in which the explicit time is shown depends on the country code specified in the CONFIG SYS file of the instrument boot disk Our sales representative or service person can set it to your preference The explicit isotope decay correction reference time can be edited with one second accuracy Although the seconds value is not shown when the explicit time is displayed it is included when isotope parameters are printed See also section 10 2 8 94 7 3 System
168. ments when the conveyor is used When measurements are made manually all active detectors can be used in measurements If an isotope has been normalized using the conveyor but an assay that uses this isotope is measured manually there may be some active detectors that cannot be used in measurement because they have not been normalized All these detectors that cannot be used in manual assay measurement are indicated in the manual assay prompt screen If an isotope has been normalized manually and after this the background is normalized using the conveyor the background values are not updated for those active detectors that cannot be used for measurement by conveyor This happens only if at least one detector has been set inactive If an assay using this isotope is subsequently measured manually the background values used for these detectors will be incorrect Therefore it is recommended that you normalize the background manually whenever any isotope is normalized manually and at least one detector is set to be inactive 92 7 3 System mode 7 3 System mode 7 3 1 System mode parameters You may change the WIZARD program operation in a number of ways Most of the parameters which affect the program operation are collected in the SYSTEM mode The following settings can be found Date Isotopes Operation mode Background Printout selections Active detectors Active hardware Diagnostic output Communication parameters Version Disk op
169. n isotope peak exceeds ThresholdChannelCounts 10 2 4 2 1 7 Checking if the coincidence peak is high enough This checking is done only if the isotope main peak height is above ThresholdChannelCounts The minimum coincidence peak height in the smoothed spectrum is given by the formula MinCoincPeakChannelCounts MinCoincHeight PeakChannelCounts 100 140 10 2 Isotope normalization and GLP TEST measurement where PeakChannelCounts is the smoothed spectrum height of the isotope main peak and MinCoincHeight SYSTEM Isotope lt isotope name gt Min coinc height If MinCoincPeakChannelCounts is less than ThresholdChannelCounts the coincidence peak is not checked it may be buried in the background and or spillover counts Otherwise if the smoothed spectrum height of the coincidence peak is less than MinCoincPeakChannelCounts the normalization is aborted 10 2 4 2 1 8Checking for unexpected peaks ThresholdChannelCounts is next increased with a counts number that is equal to the smoothed spectrum height of the isotope main peak multiplied by SYSTEM Isotopes lt isotope name gt Threshold level It is assumed that the isotope activity increases the background activity level outside the main peak area by this amount In the Diagnostic info printout the channel numbers and heights of those peaks whose smoothed spectrum height exceeds this new ThresholdChannelCounts are printed See Section 10 3 8 on page 164 If SYSTE
170. n of isotope activity with respect to isotope energy The X axis shows either MCA channel number or keV up to 1024 To toggle between these press Enter or E The selected unit is shown at the top of the spectrum The Y axis shows the accumulated counts in each individual channel The analyzer is initially adjusted so that each channel corresponds to 1 keV therefore the X axis is approximately from 0 to 1 MeV LIVE SPECTRUM 1 X scale MCA channel 8100 64 total counts in full window 76352 On the right hand side of the spectrum five functions are displayed By using the up and down arrow keys you can select the function you want A function is shown to be selected by the underline mark appearing under the function name The functions are as follows Scale This allows you to select the scale for the spectrum The default is 0 1024 By pressing the left arrow you can change the scale to 0 512 0 256 0 128 or 0 64 To return to fuller scales press the right arrow The figure shows the 0 64 scale See the next function Shift for how to move through the scale segments Shift Ifthe scale used is less than full scale 0 1024 e g 0 256 you can use the shift function to move the spectrum to see other parts which otherwise would not be displayed e g 256 512 etc Just press the right or left arrow to move in the direction you want Marker If you have the basic scale 0 1024 selected you can use this fun
171. n the counting window CPS Counts per second in the counting window Spectrum display Select the appropriate one using the right and left arrow keys Pressing the right key twice initiates the CPM display Assay Auto mode Measuring now elapsed time is 30s Det Counts per minute 01 5555 3 02 5555 3 03 12345 1 04 645 0 05 7739 2 06 6784 1 07 5479 2 08 9567 8 09 9832 8 10 9427 4 All detectors are shown simultaneously The example is from CPM counts the display for COUNTS and for CPS are analogous with the CPM display Note the CPM values are not corrected for dead time background spillover crosstalk or detector efficiency but are direct values of accumulated counts divided by measured time 2 4 3 Displaying the isotope spectrum Press the left right arrow keys four times Show cpm results Measuring now elapsed time is 29 s Show detector 1 spectrum Show detector 2 spectrum Show detector 3 spectrum Show detector 4 spectrum Show detector 5 spectrum Show detector 6 spectrum Show detector 7 spectrum Show detector 8 spectrum Show detector 9 spectrum Show detector 10 spectrum Change data with keys 20 2 Wizard controls Select the detector with the up down arrow keys then press E a spectrum is displayed For singe detector instruments there is no Spectra selection in Show cpm result instead just press E to get the spectrum The spectrum display is a graphical representatio
172. ncy depends on the total number of counts under the isotope main peak and under the coincidence peak The border between the peaks is taken to be the channel number between the peaks where the smoothed spectrum counts are the smallest If Counting window is Dynamic the low end of the main peak is taken to be the low boundary of the counting window and the high end of the coincidence peak is taken to be the high boundary of the counting window If Counting window is Dynamic keV the low end of the main peak is the first channel to the left of the main peak that is marked as Minimum this is named Peak begin channel in the diagnostic info printout The high end of the coincidence peak is the first channel to the right of the coincidence peak that is marked as Minimum this is not included in the diagnostic info printout however if such a channel is not found or the channel number would be greater than 170 the high end is taken to be 170 Because the main peak and coincidence peak windows are not the same when Counting window is Dynamic or Dynamic keV the calculated values for Horrocks efficiency may differ a little Furthermore in the Dynamic case the width of these areas depends on the value of SYSTEM Isotopes lt isotope name gt Window coverage so changing it may also affect calculated value for Horrocks efficiency The calculation uses the following formulas MainPeakCounts MainPeakCounts een
173. nded save 42 Saved files 45 Select detectors for diag info 103 Shielding 120 126 Shift sal Short cut keys 4425 Significant cpm per keV sse 96 Single peak it etos eue 96 Softkey Software Specifications ceteseds savesens in er eb e ca rage resis 125 Spectrum Bad spectr ni results un sense nee 34 Howto CHANG unseres 21 Printing via MultiCalc 198 Spectrum display sse 20 Mar Mec 21 Min Lo Win Db ases eire tr tas Spectrum type Spill determinant Spilldown Spillover Spillover correction Disable nien Stabilization Bain eee sse Standard curve files Start time STAT cunting u ne nennen ann Status line RI nennen nee Stopping an assay Store assay spectra Store spectasse asiasi eneas Stored files tte eo aereos ANE Switch on System mods rai ee T Tabulation of output fields 51 Target Values iii dias 44 Technology 185 Template sense 52 TEST label eite IR teet aa 24 Threshold level 96 95 Throughput seis 2 9 ero ep o ph Bela 119 Total CPS 4 DPS ann aan nat ciaci n 35 58 70 Transfer lane use herede nndis 102 Tray for manual mode sse 89 Trend files sess enne 57 Trends 22a laa ii ID 48 U Unkn CV flag limit
174. ngle source worst case 1 125 negligible Co 57 negligible Cr 51 negligible 126 9 Specifications Cs 137 lt 0 12 Co 58 lt 0 2 The effect of crosstalk is corrected by patented crosstalk correction software 9 5 8 Background 1 125 50 CPM Co 57 90 CPM Typical values at the factory Turku Finland background may be different elsewhere depending on local conditions 9 5 9 Energy range Default setting is up to 1024 keV 9 5 10 Gain stabilization Optimised window setting for each isotope is based on the use of multichannel analyser techniques The stability and reproducibility of the results are ensured by checking resolution efficiency spectrum drift and background 9 5 11 MCA Linear multichannel analyser with 1024 channels calibrated for the range 1 1024 keV 12 bits Analogue Digital converter Dead time max 25 us dead time effect is compensated by a program 9 6 Instrument control 9 6 1 Hardware The system is controlled by a 16 bit computer having a single 3 5 disk drive and a hard disk The computer contains 1 Mbyte of RAM memory two serial RS232C ports and one parallel port and a built in 9 screen monitor Built in keyboard with pressure sensitive keys 9 6 2 Software Operating system MS DOS v 3 21 Control by an external computer using MultiCalc immunodiagnostic data management software 9 6 3 Connections Serial ASCH interface RS 232 3 output terminals Terminal 1 for printer Te
175. normalization and you must renormalize the isotope Also the specifications are valid only with the factory settings When you select Isotopes a list of existing isotopes will be shown Select the isotope whose conditions you want to change The following parameters can be changed Note isotope parameter lines can be hidden with the Ctrl K key and made again visible with the Ctrl U key The Ctrl V makes all hidden parameter lines visible 7 3 3 1 Name This is the name of the isotope e g I 125 It is shown in the list of isotopes for normalization and assay measurement The maximum length of the name is 8 characters 7 3 3 2 Comment This is shown in the list of isotopes for normalization and assay measurement after the isotope name The maximum length of the comment is 13 characters e g lodine 7 3 3 3 Normalization time Give the time for which the isotope is to be counted if it is used for normalization 7 3 3 4 Repeat times You can set the number of times each isotope normalization measurement is repeated This means that the total time a sample is measured in each detector is this number multiplied by the normalization time that was set above The measured counts are compared with each other and the program calculates the probability that the differences between the expected and observed counts in these measurements occured It then calculates the probability that the differences between expected and observed counts in these measure
176. nt to view When it is displayed on screen four functions appear at the bottom of the display Edit 1 Press key 1 to start editing Print 2 Press key 2 to print the curve Undo C Press Clear backspace to clear all the changes made in editing and to restore the original curve to the display Exit EXIT Press EXIT to quit from curve viewing The editing operations are as follows Standard selection Use the left right arrow keys to select a particular standard on the curve Replicate selection Use the up down arrow keys to select a particular replicate point for the currently selected standard Delete 1 Pressing 1 deletes the currently selected replicate point The point which is in the form of a cross will be replaced by a square Although this point is still shown on the screen it will not be taken into account in curve fitting Ifan inserted point is deleted then it will be taken completely away Move 2 Pressing 2 followed by the up down arrow keys allows you to change the vertical position i e response value of the currently selected replicate Insert 3 After pressing 3 put the cursor at the point where you want a new point to be and press E The new point will be in the form of a cross and will be taken into account in curve fitting Undo C Pressing Clear Backspace will undo the effects of the previous editing Fit E When you press E the existing curve points will be fitted and the selection bar at the bottom of the d
177. nting 10 3 RIA IRMA RATIO assay counting When a rack having an ID clip with a numeric label in the range from 1 to 99 stuck in the area marked PROTOCOL is encountered during automatic measurement WIZARD starts measuring the assay that has this protocol number An optional rack number can be given by placing a numeric code in the RACK SPECIAL area of the ID clip otherwise the first rack is assumed to have the number and this increases by one for each subsequent rack The possible assay types are RIA IRMA RATIO and MIA The assay type is determined when an assay is created How RIA IRMA and RATIO assays are evaluated once corrected CPM values are known is not handled in this manual you may refer to MultiCalc documentation for that information Multiple isotope assays are handled in chapter 10 4 on page 171 For the 1470 counter model assay measurement can also be started manually when the conveyor or rack id readers are not used The conveyor can be disabled for the 1470 counter model by setting SYSTEM Operation mode Manual mode used to Yes and the rack id reader can be disabled by setting SYSTEM Active hardware Use rack id reader to No 10 3 1 Calculating crosstalk and spillover correction coefficients If SYSTEM Isotopes lt isotope name gt Crosstalk correction is Yes crosstalk is eliminated in RIA IRMA RATIO assay counting Spillover is always eliminated in dual label RIA IRMA RATIO assay counting In orde
178. on for WIZARD without a hard disk is V2 2B FL This version does not support the parallel port Insert the data disk into the disk drive located in the front cover of the instrument Switch on WIZARD Loading takes about three minutes before the main menu appears on the built in display Set the SYSTEM parameters Operation mode and Printout selection according to the current hardware The typical configurations are 1 Operation mode CPM or RiaCalc WIZ Serial printer SYSTEM Printout selections Use serial printer port Yes SYSTEM Printout selections Use parallel pr port No SYSTEM Printout selections Without buffering to PC No SYSTEM Printout selections Printer type Epson FX 2 Operation mode MultiCalc SYSTEM Printout selections Use serial printer port No SYSTEM Printout selections Use parallel pr port No SYSTEM Printout selections Without buffering to PC No 3 Operation mode CPM or RiaCalc WIZ Parallel printer SYSTEM Printout selections Use serial printer port No SYSTEM Printout selections Use parallel pr port Yes SYSTEM Printout selections Without buffering to PC No SYSTEM Printout selections Printer type HP PCL3 11 8 Installing MultiCalc WIZARD can be connected to an external computer running the MultiCalc or RiaCalc software The installation instructions are given in the MultiCalc manual 1224 930 During the installation procedure you wil
179. ope lt isotope name gt Norm zero time is not None The isotope half life used in decay correction is given by the parameter SYSTEM Isotope lt isotope name gt Half life hours The decay that occurs during measurement is taken into account If the reference time given by Norm zero time is Start decay correction is done with reference to the start of the normalization This normalization start time is printed in the normalization printout title line MEASURED COUNTS is the number of counts in the counting window given by WINDOW keV LOW HIGH It is not corrected in any way e g for detector dead time or background activity 154 10 2 Isotope normalization and GLP TEST measurement DETECTOR EFFICIENCY is the relative efficiency of a detector compared with other detectors It is calculated from MEASURED COUNTS which is corrected for dead time background activity and isotope decay It is printed only ifa 1470 counter has more than one detector installed RELATIVE ERROR 0 is the relative error of relative detector efficiency How it is calculated is explained in Section 10 2 16 on page 150 It is printed only ifa 1470 counter has more than one detector installed HORROCKS EFFICIENCY is the absolute detector efficiency for I 125 Horrocks efficiency is printed only if SYSTEM Printout selections Extended norm printout is Yes and SYSTEM Isotope lt isotope name gt Spectrum type is I 125 and SYST
180. ope and GLP test normalizations using the Horrock s method However you must set this parameter manually in this case This parameter can be changed without renormalizing the isotope GLP test sample DPM This value is used when absolute detector efficiency is calculated and stored in GLP data The measured sample CPM is divided with this value to get the absolute efficiency This value is not needed for I 125 since in that case the absolute efficiency can be calculated directly with the Horrocks method Maximum detector efficiency deviation In isotope normalization a check is made to ensure that the updated efficiency of each active detector differs from the average value 1 000 by at most the number of percentage points given by the value of this parameter If the difference exceeds this value for one or several detectors a corresponding warning message is printed in the normalization printout 96 7 3 System mode 7 3 3 9 Restore default isotope With this function you can copy the factory settings of any isotope to any isotope number Note this function can be found at the end of the list of isotopes To reach it quickly press End or PgDn To copy an isotope from the default list to the used isotope list use the left right arrow keys to select the isotope in the default list you want to copy Then select the isotope in the used list you want to replace Finally select Do copy and press E The isotope in the used list will be rep
181. ope peak from its expected position in a normalization measurement If the deviation of the peak position exceeds this value normalization fails for that detector Warning assay deviation This parameter gives the peak shift limit which when exceeded causes the warning Normalization recommended to be printed in assay counting Seeing this warning the user can do normalization s with the isotope s used in the assay and thus avoid any Bad spectrum messages resulting from too large peak shifts Significant cpm per keV Defines the minimum acceptable height for the peak of this isotope in the smoothed spectrum The unit is counts per minute per keV of the spectrum If no peak height in the region defined by Peak pos keV and Max assay dev or Max norm dev in the smoothed spectrum reaches this value then the activity of this isotope in the assay sample is considered to be so weak that no peak search and window adjustment is made for this isotope In isotope normalization this causes the normalization to fail for this detector Efficiency This parameter lets you set the average absolute efficiency of the detector s in the counter This enables you to calculate DPM results in addition to CPM results DPM is included in the Waste log data for isotope normalizations GLP tests and assay measurements For I 125 when Counting window is either Dynamic or Dynamic keV the absolute detector efficiency is calculated during isot
182. ops counter operation after the counting of the rack with this label An alternative is to use an empty rack as a STOP rack 2 8 2 NORM Normalization This label is used in conjunction with the isotope code label see 2 9 2 below to identify a rack as a normalization rack The isotope must be in position 10 the last position in a rack Other positions must be empty and without holders See chapter 6 for detailed instructions 2 83 BKG Background A rack with this label is used to make background normalization No sample tubes should be in the rack see chapter 6 2 8 4 TEST GLP test This label is used for coding the GLP Good Laboratory Practice test normalization The isotope must be in position 10 the last position in a rack Other positions must be empty and without holders See chapter 6 for detailed instructions Note Make sure you remove NORM BKG and TEST racks after use to avoid redoing a normalization unintentionally 2 9 Numerical labels 2 9 1 Protocol number label Protocols 1 99 are called into use by numerical barcodes the number refers directly to protocol number The label should be attached to the area marked PROTOCOL Although the ID system is used not all racks need have ID labels on them In automatic counting the racks are counted according to the last protocol number label until the next protocol number label or a special code is read 2 9 2 Isotope number code label The number of the isotope to be
183. or the particular counting conditions he or she wants Each time the conveyor presents the WIZARD ID reader with a set of samples coded with new ID the appropriate counting protocol is selected and the samples counted according to it These counting protocols can be either set up with the internal WIZARD computer or an external PC running RiaCalc or MultiCalc 8 12 Multiple label multi detector counting Dual label samples can be counted because WIZARD corrects for the spillover from one counting channel into another which often occurs with particular pairs of isotopes e g 1 125 and Co 57 A process called normalization is used to calculate this correction Once a normalization has been done for a pair of isotopes the results are stored and can be used any time that samples labelled with that pair of isotopes are to be counted The normalization process also makes possible multi detector counting by eliminating the effect of gain variation in detectors The normalization process ensure that counts for any detectors are within 1 of the mean counts of all detectors Peak positions and window settings are also optimized and background corrected for 120 8 Instrument description Another problem to be faced with multidetector counters especially with higher energy samples such as Cr 51 is that of crosstalk Radiation from a sample in one detector or on the conveyor contributes to the counts recorded in another detector Special software solves
184. ords END OF STAT ASSAY will be printed When you want to resume automatic counting press STOP and before confirming Yes end STAT remove the STAT sample tray and close the cover over the detector block The sample elevator will then be lowered to the normal position and counting will resume starting with the samples which were in the counting position when STAT was pressed Confirm Yes end STAT Note A STAT assay can also be measured in the middle of an isotope normalization but it is not possible to measure a STAT assay in the middle of background normalization 107 7 5 Power failure 7 5 Power failure 7 5 1 Introduction If power failure occurs or the instrument is switched off the microcomputer loses power all operations are aborted and the normal working memory RAM is completely cleared When power comes back the program is automatically reloaded The instrument detects that a power failure has occurred and a message Recovering from power failure is displayed a power failure message is also sent to the output devices The power failure recovery operation begins immediately Ifa further power failure should occur during the period of the first failure it will not prevent the recovery being fully completed once power returns Note Recovering from power failure can be enabled or disabled by setting SYSTEM parameter Active hardware Recover from power failure to Yes or No 7 5 2 Interrupting power failure recovery You
185. ove decay correction factor we get the corresponding value at the reference time If the isotope half life 7 is long compared to the measurement time At both the numerator and the denominator p Y g p of the formula above become almost 0 Therefore if AA lt 0 1 the formula is transformed to eT T9 e AAt where the denominator is calculated using the series expansion x AAA x ep we De n ux If an isotope normalization is done at the beginning of a 1480 multiple isotope assay MIA and the normalization has several standard replicate tubes the tube spectra are summed and the counting window is determined from this sum spectrum Likewise if SYSTEM Isotopes isotope name gt Repeat times is greater than 1 each normalization measurement is repeated as many times and the spectra are summed and the counting window is determined from this sum spectrum What decay correction factor should be assigned to the sum spectrum The value must be such that after decay correction the activity indicated by the sum spectrum is the same as the activities indicated by the individually decay corrected component spectra The formula for DCF gym the decay correction factor for the sum spectrum can be derived by assuming that all decay corrected activities are the same OM _ CTIME DOR T DCF 145 10 2 Isotope normalization and GLP TEST measurement In the above formula DCF the factor that when multiplied with the measured counts
186. ow evaluation results Operate conveyor Cpm Press START to measure 2 3 2 Function selection The line in capital letters shows four major functions of which the first OPERATE is currently selected The following lines show the commands available as part of that function Each function has its own set of commands You can select the function with the left and right arrow keys The individual command is selected with the up and down arrow keys To give the selected command to WIZARD press the E key 2 3 3 Enabled and disabled functions On the example screen shown on the previous page the first two command lines are in subdued colour This indicates that the counter is not measuring and hence these functions are not enabled The third item on the menu Operate conveyor is however available 2 3 4 Operating the conveyor If you select this and press E the display changes to Move racks to output line Move racks to input line Move both lines forwards Move both lines backwards Move all lines clockwise Rotate all lines counterclockwise Clear conveyor Press START By selecting from this menu and pressing E you can control the movement of racks on the conveyor To return to the main display press EXIT 18 2 Wizard controls 2 3 5 Selecting the mode WIZARD can be used in one ofthree modes CPM MultiCalc or RiaCalc WIZ The current mode is shown on the lower part ofthe main menu display 2 3 6 Status line At the bot
187. p of every page Numbering starts from 1 when automatic counting using the conveyor starts or stops Printer paper length WIZARD always issues a form feed when a page is full so it is important to give paper hight here Metric A4 sheet of paper is 11 inches long Valid range 1 to 99 Top margin in lines The number of empty lines at the top and the bottom of a page Valid range 0 to 99 100 7 3 System mode Paper width in inches This information makes it possible for WIZARD to select a smaller font in order to fit a whole line to the page Metric A4 sheet of paper is 8 inches wide Valid range 1 to 99 Left margin in inches Use this parameter to reserve space for left margin Note that for HP LaserJet printers a change in the value of this parameter becomes effective only after a new page has been printed You can do this by printing e g the latest background measurement SYSTEM Background Print background Valid range 0 0 to 10 Small text SLPI If Yes 8 lines of text per inch are printed If No then 6 lines of text per inch are printed Small graphics If Yes graphical sections of the printout are printed smaller with greater resolution This 1s convenient 1f the printer is able to print high resolution graphics Output negative CPM as 0 If Yes negative CPM values are output as 0 CPM In this case also negative CPM results to RiaCalc WIZ or MultiCalc are replaced with 0 values The
188. pe is I 125 its coincidence peak is searched for as explained in Section 10 2 4 2 1 5 on page 140 Then the main peak of the other isotope is searched for again among the remaining peaks in the interval where the highest most active peak is assumed to be the main peak for that isotope If this other isotope is I 125 its coincidence peak is also searched for For both isotopes the threshold level is calculated separately and the height of the isotope peak is checked against it as described in the preceding paragraph for the single label case Unexpected peaks are searched for if for neither isotope Spectrum type is Many peaks The level that a peak height must exceed in order for the peak to be considered an unexpected peak is the sum of the individually calculated threshold counts values for both isotopes If at least one unexpected peak is found the spectrum is considered bad In this case a note about this is included in the assay printout and the corrected CPM values are set to 0 If both isotopes exceed their threshold counts levels the program makes one more check to ascertain that one of the peaks is not a spillover peak from the spectrum of the other isotope If at least 1 3 of counts in either window are spillover counts then that peak channel number is considered unreliable and the expected peak channel number is used instead 10 3 3 Calculating corrected CPM values Once the effective gain value has been obtained the counting window
189. peration with this mode 1s described in Part 3 of this manual RiaCalc WIZ Data evaluation is carried out in WIZARD with the RiaCalc WIZ software See Part 4 MultiCalc Data evaluation is carried out in the PC using MultiCalc immunodiagnostic data management software See Part 5 You can operate WIZARD as an automatic or manual counter with any of these three evaluation modes assuming the appropriate options are installed on your WIZARD The change from automatic to manual is accomplished by selecting Manual mode used YES Sample elevator is raised automatically ER sure samples are loaded according to numbers on tray 89 7 2 Manual operation Note the change to manual mode cannot happen while the cover over the detector block is open This is because the change to manual mode causes the raising of the arm which transports samples from the conveyor to the detector block to make space for the sample tray to be loaded For safety reasons this movement cannot happen without the cover being closed Afterwards the cover must be opened for accessing the detector block Save the changes Now you can exit the SYSTEM mode and go to the OPERATE mode 7 2 4 Loading samples Put samples into the wells in the sample tray Each well is marked with the number of the detector which will be used to count it Make sure you load samples according to these numbers so that output results are in the right order Move the sample tray to the de
190. ps etc Choose the level that most suits you You do this by pressing the softkey F7 lt LEVELS and then the appropriate LEVEL softkey F1 F5 3 Check that microfloppy named 1470 Datadisk is in the WIZARD disk drive If not place it there so that the name label is upwards Switch on WIZARD After about 3 min the display will show 1470 Main Me Ud PROTOCOL FILES SYSTEM Sb show cpm results Operate conveyor MultiCalc Press START to measure Check the time and date by selecting SYSTEM mode and DATE If it is not correct then give the correct value See System Time amp Date setting Press EXIT to return to the main display Make sure that the counter is in the right mode i e that the text MultiCalc is showing in the lower part of the screen as in the example here If it is not select SYSTEM and then the operation mode The RIA evaluation must be MultiCalc Now you are ready to start MultiCalc assay protocol operation of WIZARD Note The HELP command given to the instrument in MultiCalc mode from a terminal connected to the PC port e g MultiCalc terminal gives extensive information about communication with external MultiCalc This information is also in the file COMMHELP TXT in the instrument hard disk 64 5 Operation with external MultiCalc 5 3 MultiCalc protocol MultiCalc is controlled by protocols A protocol is a list of parameters which need to be set or given values e g protocol
191. r is saved This means that in the case of a ten detector instrument 10 x 10 spectra are obtained and saved After this the first time you start to count samples labelled with a particular isotope or combination of isotopes the appropriate crosstalk and spillover correction factors are calculated and saved and then used in the actual sample counting When the same isotope or combination of isotopes is counted again the correction factors already saved are used 78 If you do a normalization with a particular isotope any already existing correction factors calculated using the previous normalization made with that isotope are deleted from the memory but the actual spectrum information for other isotopes is not affected so other normalizations do not need repeating New correction factors are then calculated when needed taking account of the new normalization If a background normalization is made or if the number of active detectors is changed all saved correction factors will be deleted and new ones calculated when needed If you change the crosstalk selection on or off for a particular isotope any correction factors involving that isotope will be deleted so that new ones can be calculated when needed This system has the advantage that you do not need to do special dual label normalization because the instrument can calculate the necessary dual label information based on the single label information already stored It also means that you c
192. r to do this correction coefficients must be known 10 3 1 1 Single label counting The correction coefficients are determined during isotope normalization as described in Section 10 2 13 on page 147 10 3 1 2 Dual label counting Correction coefficients have to be calculated for each isotope pair every time either one of the isotopes is normalized Since it is probable that only a few isotope pairs are used in dual label counting it is practical to calculate the coefficients the first time a dual label assay using such an isotope combination is used after normalization of either of its isotopes The coefficients are then stored in the instrument hard disk 157 10 3 RIA IRMA RATIO assay counting ASSAY 22 Sep 1994 10 44 43 Protocol id 29 PROTO1 Time limit 30 Count limit 99999999 Dual isotopes I 129 Co 57 Protocol date 22 Sep 1994 10 43 52 Run id 10 1 129 00 57 POS RACK DET BATCH TIME COUNTS CPM ERROR COUNTS CPM ERROR 1 1 1 1 30 356 690 9 6 25 21 14 3 82 86 2 1 2 1 30 3343 6602 8 2 48 30 10 7 121 55 3 1 3 1 30 3973 7917 0 2 33 22 7 3 160 70 4 1 4 1 30 348 673 0 6 30 35 30 9 43 62 5 1 5 1 30 4184 8386 3 2 28 24 2 7 438 12 6 1 6 1 30 3485 6971 9 2 43 33 8 1 170 84 7 1 7 1 30 5730 11453 6 2 06 26 6 5 196 25 8 1 8 1 30 8527 17077 2 1 82 37 16 3 88 06 9 1 9 1 30 13752 27522 9 1 59 34 1 1 64 43 10 1 10 1 30 278 23 6 6 72 80956 164663 4 0 67 11 2 1 2 30 0 0 0 0 00 0 0 0 00 Error Bad spectr
193. ratory Your WIZARD may have 1 2 5 or 10 detectors and be able to take 550 or 1000 samples at a time depending on the model you have Every WIZARD has its own built in display and keyboard for full communication with its users WIZARD can operate as an automatic standalone CPM counter or it can be used to do extensive data evaluation with its own internal RiaCalc WIZ program or it can be linked up to an external PC and use the power of a MultiCalc program RiaCalc WIZ includes the built in hard disk WIZARD can also be used as a manual counter if you require 1 3 Introduction to this manual 1 3 1 Step by step instruction sheets There are three sheets following this page which give a brief step by step outline of how to use WIZARD in each of the three operating modes 1 WIZARD is used as a stand alone CPM counter Counting parameters are set in the counter CPM results are sent to the built in display and a printer 2 WIZARD is run with RiaCale WIZ All counting protocols are set in the counter itself Results are sent to the built in display and a printer 3 WIZARD is connected to a personal computer running MultiCale software All counting protocols are set in the personal computer Results are automatically returned to the PC for final evaluation and output More information about MultiCalc will be found in the MultiCalc User Manuals You should find enough information on the appropriate one of these sheets for normal operation in any of
194. re going to use in your measurement See chapter 6 Normalization 3 4 Protocol editing 3 4 1 What is a protocol A CPM protocol contains three parameters for controlling counting conditions Before counting can be done a protocol must exist Each protocol must have a name and an ID number This number allows the protocol to be called into use by means of a barcode label To access protocol file handling you must first select the PROTOCOL option on the display 29 CPM operation 3 4 2 Protocol file handling There are several commands available to use with a protocol the most common ones are Create making a new protocol and Edit change the contents of the existing protocol The list that appears after you select PROTOCOL is as follows 1470 Main Menu OPERATE lYNezkeleie FILES SYSTEM Submenu Create Copy Rename Delete Recover Print Load More 4 Press START to measure Create Select Create and press E Give the protocol name There is a default name of the form Protnn where nn is the protocol number If you want to change this name you must use the external keyboard unless you only use numbers for the name A maximum of eight characters can be given Select an unused ID number between 1 to 99 Press E The protocol parameters can be set in a similar way to those in editing Edit Select Edit and press E Select the protocol by name from the list of protocols available and press
195. response to the Select detectors for info parameter 7 3 9 1 Print diagnostic info Diagnostic information is printed for all those detectors that have been selected with the parameter Select detectors for info 7 3 9 2 Select detectors for diagnostic information This parameter lets you select those detectors for which special information is desired 7 3 9 3 Assay repeat times The value of this parameter specifies how many times each batch of an assay is measured before proceeding to the next batch This function is used when the counter is tested for stability 7 3 9 4 Spill determinant The determinant is included in the output if this parameter YES Normally the value of the determinant should be near 1 0 Ifthe value is near zero the corrected CPM values can be inaccurate See also sections 10 3 1 and 10 4 1 7 3 9 5 Print dead time factor To do this set the parameter Yes The CPM printout is sent to a printer that has been connected directly to the WIZARD printer port when the Operation mode is CPM or you are saving RiaCalc WIZ results on a datalogger If you are using WIZARD with MultiCalc set Print dead time factor to No See also section 10 1 3 7 3 9 6 Print measurement start time The exact time when measurement of each sample was started can be included in CPM printouts and datalogger files The time is given to the nearest second If this starting time output has been selected a field named CLOCK appears at the rig
196. rgy counts that have energies exceeding the range of the MCA they are also reported in the total background CPM The open window background counts value is corrected for detector dead time and then converted to CPM The conversion from counts to CPM is done with the formula 60 Counts CountingTimelnSeconds Sometimes isotope activity is expressed in CPS counts per second The conversion from counts to CPS is done with the formula CPM CPS Counts CountingTimelnSeconds 10 1 3 Dead time correction Dead time correction compensates for counts lost due to the finite counting rates of the detector and the measuring circuit For 1480 the following formulas are used DeadTimeCorrectedCPM DeadTimeFactor MeasuredCPM T 6 45 10 1 DeadTimeFactor EBEN 1 CPS T CPS T 131 10 1 Background normalization Here CPS is the counts per second activity and t is the dead time of the measuring circuit in seconds for each measured count The formula takes into account that at high sample activities pulses pile up so that several pulses are detected as one pulse with higher energy A different formula for calculating the dead time factor is used in 1470 For this counter model up to five detectors can be connected to the same analogue to digital A D converter A 10 detector counter has two A D converters Because of this the dead time factor of one detector is affected by counts from other detectors that are connected to the same
197. rial printer EE 2 PC RS 232 port to connect the PC 3 PRINTER PARALLEL Parallel port to connect a parallel printer HP PCL 3 The parallel port is standard in new instruments where the software version V3 5 is factory installed The third communication port in older instruments is a RS 232 serial port and is marked as MAINFRAME V3 5 does not support the mainframe port The printout selection Use parallel pr port is available only if the microprocessor board has a parallel port DCE CPU board WARNING Do not in any circumstances connect a parallel printer to the RS 232 serial port i e to the Printer serial PC or Mainframe port This may damage the parallel printer The parallel printer can be connected only to the port marked as PRINTER PARALLEL SERIAL 1 PRINTER 1 EPSON FX CPM or RiaCalc WIZ Pera printer 1221 502 WIZARD CONNECTION PANEL gt 2 1 PRINTER J E EL wo MultiCalc ib 2PC P 3 PRINTER PARALLEL PARALLEL 3 PRINTER 1224 122 1 HPPCL3 CPM or RiaCalc WIZ Parallel printer Loan m Plug in the power cables for each device Switch on the printer and PC 185 11 Installation 11 7 Switching on WIZARD If you have WIZARD without a hard disk you must insert the program disk into the drive located in the microprocessor rack This disk remains in place permanently The latest software versi
198. riodically tested to ensure optimum counting conditions i e that the optimum efficiency and background ratio is achieved WIZARD makes all these corrections automatically based on the information obtained during normalization This procedure involves measuring a single sample in each detector in turn The instrument must be normalized for each isotope used Normalization can be made infrequently When you have installed WIZARD normalize it for the isotopes you are going to be using Unless the instrument gives a warning asking for a normalization a time period of six months before you redo the normalization is appropriate Once you have made a normalization you can then create or edit a normal counting protocol where you can select the isotope you have normalized and use it to count samples If the instrument is not normalized for a particular isotope and at some time you try to run an assay with this unnormalized isotope selected WIZARD will give an error message telling that the isotope is unnormalized and will terminate the assay The procedure for doing background and isotope normalization is explained in the following sections 6 1 2 Background normalization A background normalization is a method of determining the background for each detector It is important that 1t has been made at least once The background value is subsequently subtracted from count values during actual counting To make a background normalization stick the ID
199. rminal 2 for system PC external MultiCalc Terminal 3 for parallel printer 9 7 Electrical safety requirements The design of the instrument is based on the following electrical safety requirements EN 61010 1 1993 IEC 1010 1 CSA C22 2 No 151 1986 CSA C22 2 No 0 4 1982 CSA C22 2 No 0 M91 EN 50082 1 1992 EN 50081 1 1992 127 9 Specifications 9 8 Isotopes defined for 1470 WIZARD ID Isotope Name Energy Eff Halflife Coverage Low W High W Res keV hours keV keV 1 I 125 lodine 29 82 1445 97 25 2 Co 57 Cobalt 122 90 6480 92 13 3 Cr 51 Chromium 320 3 7 667 80 9 4 1 129 lodine 31 65 1 49E 11 96 24 5 As 76 Arsenic 559 7 26 4 31 6 Au 195 Gold 99 75 4390 95 T Au 198 Gold 412 11 64 7 47 8 Ba 133 Barium 356 16 6 30E 04 54 9 Ba 139 Barium 166 76 1 38 87 0 Br 77 Bromine 245 11 57 74 2 Cd 109 Cadmium 22 71 11136 16 32 3 Ce 141 Cerium 145 56 780 125 167 4 Co 58 Cobalt 810 4 1711 660 930 5 In 111 Indium 416 42 67 7 150 500 6 Cs 134 Caesium 795 30 18063 25 500 890 10 7 Cs 137 Caesium 662 26 2 63E 05 62 8 Er 171 Erbium 308 26 7 52 9 F 18 Fluorine 511 8 1 83 85 20 1800 21 Ga 67 Gallium 185 70 78 22 Gd 153 Gadolinium 147 100 5808 68 26 167 23 Hg 203 Mercury 279 31 1126 88 24 1 123 lodine 159 80 13 3 10 25 1 131 lodine 360 15 193 260 430 26 In 111 Indium 416 42 67 7 320 541 27 In 114m Indium 190 42 1188 52 166 210 29 K 43 Potassium 373 14 22 6 37 30 Na 22 Sodium 511 8 2 27E 04 31 Nb 95 Niobium 766
200. rom several separate measurements The error calculation routines are not affected by this The individual repeat measurement CPM s that are used to calculate the Chi Square probability are determined from each repeat measurement s own spectrum using the counting window that has been determined from the sum spectrum An isotope normalization cannot have repeat measurements if it is done at the beginning of a 1480 multiple isotope assay 151 10 2 Isotope normalization and GLP TEST measurement 10 2 17Isotope normalization printout Here are some printout examples NORMALIZATION OF 4 I 129 26 Aug 1994 09 59 14 Total counting time 45 Repeat times 3 Nominal gain 1 00 keV channel Main peak 31 0 kev Main peak at nominal gain 31 0 channels Nominal window coverage 96 00 PEAK PEAK RESOL WINDOW keV DECAYED MEASURED DETECTOR RELATIVE SIGNIF DET CHN DEV LOW HIGH ACTIVITY COUNTS EFFICIENCY ERROR LEVEL 1 32 2 3 8 21 0 16 9 46 5 1 0000 106501 1 0008 0 28 44 494 2 32 5 5 0 19 2 18 6 46 0 1 0000 106565 1 0014 0 28 93 265 3 32 6 5 1 21 3 17 6 45 9 1 0000 107351 1 0090 0 27 6 074 4 32 5 4 8 22 0 17 7 46 1 1 0000 106443 1 0002 0 28 76 265 5 32 1 3 6 21 9 16 9 46 5 1 0000 106904 1 0047 0 28 48 776 6 32 8 5 8 20 4 17 5 45 6 1 0000 106199 0 9979 0 28 44 679 7 32 2 3 8 20 6 17 8 45 6 1 0000 107295 1 0084 0 27 43 328 8 32 7 5 5 21 1 17 5 46 6 1 0000 106034 0 9963 0 28 85 655 9 31 9 2 8 21 9 17 0 46 9 1 0000 106420 0 9999 0 28 25 735 10 32 4
201. rotocols themselves rather than just individual parameters in a protocol e g copying a protocol or ordering the list alphabetically etc you can do this by using the appropriate softkeys see the User Manual 5 5 Running an assay 5 5 1 Starting the run Make sure that the first cassette has 1ts correct protocol selection ID For more information about this see ID system in Part 2 of this manual Put a STOP ID on the last sample rack to be counted or use a STOP rack or a totally empty rack in order to stop the instrument automatically Load racks starting with the right hand conveyor lane Select the MultiCalc main menu Press COUNTER softkey F1 key Select 1470 and press ENTER MultiCalc will now store all protocols in the instrument automatically and the instrument will count all the loaded samples 5 5 2 Stopping the run The counting will stop automatically when a STOP rack or ID is found You can also press the STOP key on the WIZARD keyboard In this case the following text will appear on the built in display Continue End assay continue End assay Clear conveyor Depending on whether you want the next assay to be counted or all counting to stop select End assay continue or End assay clear conveyor respectively To override the stop instruction and continue counting select Continue 68 5 Operation with external MultiCalc 5 5 3 Result handling There are a number of options concerning the way the re
202. rtant 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 11 3 Electric power Three electrical outlets each with 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 voltage are anticipated a mains stabilizer may be necessary 11 4 Unpacking Unpack all units and accessories and check them according to the packing list also noting any possible transport damage Move the instrument to its place of operation Remove the rear panel of the instrument by loosening the captive screws Remove the two plastic bands which keep the elevator at the lowest position during transportation see the figure below REMOVE THESE BANDS m Xo x gt m BH lt _ 4 j XL Jo N gt Qu DON c oo lt 5 J 5 O y fa B lt 183 11 Installation Remove the front lower cover by unscrewing the screws at the front of the instrument see the figure below Check that all the boards in the electronic racks are firmly in their positions Check also all the cable connec
203. s The number of replicates for each isotope standard can be set independently At the beginning of a multiple isotope assay isotope standards are normalized in order to determine counting windows and spillover For each standard sample the replicate tube spectra are summed and this sum spectrum is used when that isotope is normalized see Section 10 2 3 on page 136 The replicate number for a standard can also be set to 0 This means that there are no tubes for this isotope standard in the multiple isotope assay and the latest previously done normalization for that isotope is used instead 10 4 1 Spillover correction If spillover correction has been specified in the MIA protocol the spillover correction factors are calculated in the following way During isotope normalization the spectrum of the normalization sample is stored in the instrument hard disk Latest background spectra are also copied and stored with the normalization spectra The program determines counting windows in terms of MCA channels This is done by converting the stored keV values of counting window boundaries by using the effective gain during the isotope normalization during which the counting window was determined This is discussed further below If SYSTEM Isotopes lt isotope name gt Counting window is Dynamic the window boundary keV values are determined during isotope normalization on the basis of the parameters SYSTEM Isotope lt isotope name gt Windo
204. s and average results are printed out You may switch off or on values shown in the list by selecting NO or YES respectively ES E ES E E F ES ES E ES E ES E ES E ES E ES E ES E ES E Individual standard values Replicate standard averages Dilution standard averages 2 fields after standards Individual unknown values Replicate unknown averages Dilution unknown averages 2 fields after unknowns Individual control values Replicate control averages Dilution control averages ao as e e K Dilution averages effects only if samples are divided into separate dilution groups 2 fields are those which would be the same for every item By means of a switch you can tell the program to only output such fields once after standards or unknowns respectively 4 6 10 Copy settings from template This parameter is to allow you to reuse a particular set of sample result outputs without having to enter each one individually You have the choice of selecting the Display sample result output or the output saved in one of three templates Single label RIA IRMA output i e SEQ PAT CODE TIME CPM CONC UNIT CV FLAG Dual label RIA IRMA output i e SEQ PAT CODE TIME CPM CPM B CONC CONC B UNIT CV CV B FLAG 52 4 Operation with internal RiaCalc WIZ Single label RATIO i e SEQ PAT CODE TIME CPM 4 7 Leaving the editor Press EXIT to leave the editor A choice of three possibilities is sho
205. s A MB Then the matrix M is inverted Let us denote the inverted matrix with M When this is multiplied during assay counting with a vector consisting of the measured CPM values in all isotope counting windows the result vector consists of spillover corrected isotope activities that is B MA If the matrix M cannot be inverted the message The spill matrix could not be inverted No spill correction is made is printed If the absolute value of the determinant of the matrix M is smaller than 0 1 the message Results for CCPM DPM and RATIO may be inaccurate is printed 1 72 10 4 Multiple isotope assays MIA MULTI ISOTOPE ASSAY 10 PROTO1 STANDARDS NORMALIZATION OF 1 I 125 04 Oct 1994 14 53 59 Total counting time 600 Nominal gain 1 00 keV channel Main peak 29 0 kev Main peak at nominal gain 29 0 channels PEAK PEAK RESOL WINDOW keV DECAYED MEASURED HORROCKS STANDARD CHN DEV LOW HIGH ACTIVITY COUNTS EFFICIENCY CPM 22 9 21 1 29 8 15 0 35 0 1 0000 417276 84 1 42115 E SAVED E OF ISOTOPE NORMALIZATION NORMALIZATION OF 2 Co 57 04 Oct 1994 14 54 06 Total counting time 600 Nominal gain 1 00 keV channel Main peak 122 0 keV Main peak at nominal gain 135 0 channels Nominal window coverage 92 00 PEAK PEAK RESOL WINDOW keV DECAYED MEASURED STANDARD CHN DEV LOW HIGH ACTIVITY COUNTS CPM 100 9 25 2 16 2 92 2 177 1 1 0000 1621968 165162 VALUES SAVED END OF ISOTOPE NORMALIZATION NORMALIZATION OF 3 Cr 51
206. s failed In detector 9 a peak due to Compton scattering was so high that is was regarded as an unexpected peak The normalization succeeded when SYSTEM Isotopes Cr 51 Threshold level was raised from 20 to 30 Total counting time is the time a tube is measured in one detector If SYSTEM Isotope lt isotope name gt Repeat times is greater than 1 the counting time printed is this number multiplied with SYSTEM Isotope lt isotope name gt Normalization time Repeat times is the number of repeat measurements given by SYSTEM Isotope lt isotope name gt Repeat times It is printed only if it is greater than 1 Nominal gain is printed only if SYSTEM Printout selections Extended norm printout is Yes For the 1470 and 1480 normal energy ranges it is equal to SYSTEM Active hardware Nominal gain The factory setting for 153 10 2 Isotope normalization and GLP TEST measurement it is 1 0 keV channel If the value of this parameter is changed it affects only how keV s are converted to channels and vice versa by the software not the effective gain of the electronics circuit The latter must be set by a service technician For the 1480 model the parameter Nominal gain is the normal energy range gain The nominal gain for the 1480 extended energy range gain is 2 0 keV channel and cannot be changed Main peak is the value of SYSTEM Isotope lt isotope name gt Peak pos keV It is printed on
207. s forward Position 1 when loading racks 2 Load racks onto WIZARD Make sure racks are loaded the correct way round with the ID clips facing away from you as shown in the figure Start by loading the right hand side of the conveyor 3 Press F1 F COUNTER select 1470 and press Enter Live display during counting is obtained by selecting from the main menu on the WIZARD built in display Operate and then Show cpm results Available live displays are counting parameters Counts CPM CPS and Spectrum These appear on the built in display Live display available during counting MultiCale is a very versatile data handling program which runs on an external computer You can use it to make counting protocols These will be transferred to WIZARD to control counting Results are returned to MultiCalc for evaluation They can also be sent via MultiCalc to a local area network LAN or mainframe computer Output to external computer e g a main frame Connection is from the mainframe to the PC Results go to the PC connected to port 2 Printout goes to the printer connected to the PC For System setting info e g detector de activation or clock setting select SYSTEM in the main menu 13 2 WIZARD controls 2 Wizard controls 2 WIZARD controls 2 1 Introduction This chapter describes features you need to use to control WIZARD These features are the one or two keyboards the display the ID sy
208. sample s response value with a BLANK BLANK NSB Non Specific Binding The commonly used responses are B BO and LOGIT T is the total sample response value In RIA it is presumed that T gt BO gt X gt BLANK where X is the response of any unknown or control value 4 5 7 Fitting algorithm Choose the fitting algorithm for the standard curve from the list LinInt linear interpolation LinUwReg linear unweighted regression ParUwReg parabolic unweighted regr CubUwReg cubic unweighted regression LinWgReg linear weighted regression ParWgReg parabolic weighted regr CubWgReg cubic weighted regression InSpline interpolated spline AuSpline auto smoothed spline SmSpline manually smoothed spline 4 5 8 Std outlier reject Select NO if there is to be no check for outliers but if you select CONDITION then two more parameter lines appear Diff from mean 20 and 400 Diff from curve 10 The first specifies that an outlier will be rejected if its percentage difference or the absolute difference from the mean of the replicates equals or exceeds the value given here The second uses a standard curve as a reference instead of the mean You can also specify whether the larger or smaller difference of two replicates is to be the deciding factor in the outlier rejection 4 5 9 Curve edit halt Choices are YES halt for curve edit NO do not halt for curve edit REF CURVE evaluate using reference curv
209. seen by choosing the Show evaluation results function The built in program allows counting and normalization protocols to be created and edited The built in display and keyboard is used Results are sent to the display and a printer For System setting info e g detector deactivation or clock setting select SYSTEM in the main menu The printer connected to WIZARD port 1 is used for printing corrected CPM results directly from WIZARD A disk can be used for transferring results from WIZARD External keyboard in built in drawer for extended protcol editing 11 MultiCalc Operation of WIZARD 1 Fix ID clips to racks Fix ID clip here Pax ID labels barcodes are stuck to an ID clip which fits onto a rack to tell WIZARD the function of the rack A counting protocol is a set of three parameters time max counts limit and isotope which control counting Rack number is optional and allows each rack to have its own number Normalization ensures that the counting efficiency of each rack sreca rmorocoL Protocol number or isotope detector is the same number Background ensures that the effect of the background is removed y i Rack number or NORM from the measured counts HO MM TEST BKGorSTOP Test initiates a GLP performance test normalization redon Isotope number shows the isotope to be used in normalization Stop tells WIZARD that no more racks are to be counted ID clip face
210. separated by tabulator characters This makes it easier to read several spectra at the same time into a spreadsheet program 69 5 Operation with external MultiCalc Ortec format This binary file format is called Integer Data File in Ortec documentation The file extension 1s CHN Each spectrum is stored in a file with the following path and name A EVAL lt assay name gt E lt run id number gt lt assay position gt CHN run id number gt can take the values 00 01 02 99 and assay position gt the values 000 001 002 999 If FILES Spectra Operation is Send to PC then all spectra files are sent at the same time one after another and the receiving end must separate them apart 5 6 2 GLP data This allows you to handle the GLP data obtained in GLP normalization There are four options some of which lead to other options Operation View allows you to view the GLP data Delete delete GLP data Criteria allows you to select the warning limits for different types of GLP data You can select Print criteria Item many items appear from which you can select the one which you want the GLP data PEAK BGRD EFFIC RESOL EFFICIENCY COVERAGE CHI PROB WIN CPM TOTAL CPM Isotope you can select the isotope type from those for which a GLP normalization can be done Detector Do operation 5 6 3 Waste log file A waste log file that contains the total CPS and DPS values o
211. serial ports described above WIZARD can write results to a floppy disk that is loaded into the drive in the instrument front panel The disk should be a 3 5 high density 1 44 MB microfloppy disk and it must have been formatted before using This SYSTEM parameter allows the datalogger feature to be switched on or off If YES is selected then CPM results are saved on floppy disk and can be later evaluated in a PC In particular you can use the MultiCalc program to evaluate WIZARD result files in the desired order direct from the floppy disk The results are saved in a subdirectory of the data disk called RESULT All assays are put in their own files The file names used are as follows Unknowns AXX_ YYY T where XX is the protocol number and Y YY is the assay order Background norm BKG__YYY T where YY Y is the assay order Isotope norm NZZ__YYY T where ZZ is the isotope number and Y Y Y is the assay order GLP test normalization TZZ_YYY T where ZZ is the isotope number and Y YY is the assay order Examples BKG__001 T NO1__002 T NO2__003 T A01 004 T A01__005 T A02 006 T 99 7 3 System mode The assay order number starts again from one after 999 so it is possible that a file with same name already exists In this case the old result files are overwritten The extension of the already handled result files should be changed to ZT to prevent this happening If the data disk gets full WIZARD automatically tries to delete all the files with
212. sotope normalisation only results are stored differently Data obtained in a GLP TEST normalisation is not used in assay measurements but is tested against preset limits and then stored so that it can later be compared with other TEST normalisations using the same isotope This comparison is done by presenting the values of some measured parameters as a function of time so that any systematic trends or large random deviations can easily be discerned 121 9 Specifications 123 9 Specifications 9 Specifications 9 1 Physical dimensions Height 560 mm 22 0 Width 650 mm 25 6 or 1170 mm 46 1 Depth 770 mm 30 3 or 650 mm 25 6 Weight 128 kg or 140 kg The smaller dimensions are for the 550 sample version and the larger for the 1000 sample version 9 2 Electrical requirements 230 V 10 at 50 60 Hz 115 V 10 at 50 60 Hz 200 VA maximum 9 3 Environmental requirements Temperature range from 15 C to 35 C Max humidity 85 9 4 Sample handling 9 4 1 Sample vials Max diam 13 mm 17mm in manual mode without a tray Max cap diam 14 mm Min diam no limit Max height 90 mm including cap Vial shape no shape restrictions 9 4 2 Sample racks Plastic racks 10 samples rack Length 164 mm Width 18 mm Height 57 mm Max centrifugation force 2500 G Max temp 40 C Racks are provided with individual sample carriers which can be replaced in the case of contamination 9 4 3 Sample changer
213. spectra of MIA standard replicate measurements If an isotope standard normalization is done at the beginning of a 1480 multiple isotope assay MIA the isotope standard sample can consist of several replicate tubes These are all measured and their spectra are summed When the isotope counting window has been determined from the sum spectrum the individual replicate spectra are used to determine the counts CPM and relative activity of each standard replicate tube These are included in the isotope normalization printout Otherwise all other normalization results are determined from the sum spectrum 10 2 4 Isotope counting window In order to sum counts over an isotope counting window its boundaries must be known in terms of MCA channels 136 10 2 Isotope normalization and GLP TEST measurement If SYSTEM Isotope isotope name gt Counting window is Fixed or Dynamic keV the ke V values of the boundaries are given by the parameters SYSTEM Isotope lt isotope name gt Low boundary keV and High boundary keV and they only need to be converted to MCA channels If Counting window is Dynamic the counting window is first determined in terms of MCA channels and these are later converted to keV s They are again converted to MCA channels during assay counting using the effective gain at assay counting time Counting windows in terms of MCA channels are needed during isotope normalization to calculate the following data
214. ssays The string is not included in files that are stored to datalogger disk and it is sent to MultiCalc only if results are not buffered 7 3 9 9 Read spectra from diskette To allow convenient testing of the data reduction program in WIZARD spectra can be read from a text file in the datalogger disk instead of from the multichannel analyser Special commands make it easy to write artificial spectra that have just the properties you want However you can also use real assay spectra if you want e g to test how different isotope normalization settings affect assay results The application note Testing WIZARD with artificial spectra describes the format of this text file so that you can write your own tests When this parameter is Yes the text Reading spectra from diskette appears in the main menu status area When measuring normal samples be sure to set this parameter to NO otherwise all counts values will be zero 7 3 9 10 Disable dead time correction Dead time correction can be switched off for testing purposes by setting this parameter to YES The default is NO See also section 10 1 3 7 3 9 11 Disable spillover correction If this parameter is set to YES spillover correction is not made in dual labelled RIA IRMA RATIO assays If spillover correction is disabled then so is also crosstalk correction even if isotope parameter Crosstalk correction is YES See also section 10 3 1 7 3 9 12 Disable background correction I
215. ssment is Bad spectrum Expected peak chn is in isotope normalization the main peak set by SYSTEM Isotope lt isotope name gt Peak pos keV converted to a MCA channel number using the nominal gain If an isotope uses a fixed window then expected peak channel is not used 167 10 3 RIA IRMA RATIO assay counting In RIA IRMA RATIO assay counting the expected peak channel is the MCA channel number where the isotope peak was in the latest batch where it was sufficiently high The peak position is saved at the end of each assay and if the parameter SYSTEM Operating mode Default is norm window is No the expected position is retrieved at the beginning of next assay using the same isotope Ifthe parameter Default is norm window is Yes the peak determined at isotope normalization time is taken as the expected peak position at the beginning of an assay The expected peak channel is used as a substitute for the observed peak channel if the current tube s activity is so small that the isotope peak does not exceed the threshold level see Section 10 2 4 2 1 6 on page 140 The isotope peak in turn is used to calculate the effective gain if SYSTEM Isotope lt isotope name gt Counting window is Dynamic keV or Dynamic The gain value is used to convert counting window boundaries from keV to MCA channel numbers In 1480 multiple isotope assays MIA the windows determined at isotope normalization time are alw
216. statistical nature of radioactive decay and other errors in measurement can cause the CPM values that have been corrected for background relative detector efficiency and crosstalk in multi detector counters and spillover in multiple labelled assays to be negative Max assay run id When several assays having the same protocol id number are measured one after another this parameter determines how many RiaCalc WIZ data files and assay spectra files are saved before new ones overwrite the older The assays are differentiated by the run id number The run id number is increased by one for each new assay having the same protocol id number When the Max assay run id has been reached the next run id number starts from 1 again The value for Max assay run id can be from 2 to 99 Setting the value to a small number limits the number of RiaCalc WIZ data files and assay spectra files that are stored in the instrument hard disk It also limits the number of RiaCalc WIZ data files that are loaded from diskette Note that the parameter SYSTEM Printout selections Max assay run 1d affects only new run id numbers that are being created during automatic measurement You can still view copy from and to and delete files with greater assay run id numbers than Max assay run id Also when this parameter is made smaller no files are deleted but you can always manually delete files with greater run id numbers 7 3 7 Active detectors
217. stem and the HELP function When you understand how to use these features then you can proceed to the following chapters to see how to use WIZARD to get the results you want 2 2 Keyboard WIZARD can have two keyboards a simple membrane type on the front of the instrument see the figure below and a second a complete PC keyboard which is in a separate compartment The instrument is normally operated using the small keyboard whereas the larger keyboard is used for extended protocol editing because the small keyboard does not include letters and certain other characters which are needed in some editing operations The keyboards are connected in parallel with each other and each key on the built in keyboard has its equivalent on the external one The keys on the external keyboard which correspond to those on the built in keyboard are as follows Built in keyboard External keyboard START F3 STOP FA STAT F5 HELP F1 EXIT ESC C Clear Backspace E Enter Use the EXIT ESC to escape from the operating level the program is on and go back to the previous one If you are using a PC running MultiCalc all commands involving MultiCalc are given via the PC keyboard 17 2 Wizard controls 2 3 Display 2 3 1 Main menu display When you start to work with the instrument you will see on the built in display something like 1470 Main Menu 9 ud V Vis PROTOCOL FILES SYSTEM Submenu Show cpm results Sh
218. stored on the instrument hard disk For the 1470 counter model the background measurement can also be started manually when the conveyor or rack id reader are not used The conveyor can be disabled for the 1470 counter model by setting SYSTEM Operation mode Manual mode used to Yes and the rack id reader can be disabled by setting SYSTEM Active hardware Use rack id reader to No For the 1480 counter model background is measured for both the normal and the extended energy range if there is no label stuck in the PROTOCOL area of the ID clip If an additional odd number ID label is in the PROTOCOL area the background is measured only for the normal energy range 15 1000 keV An even number label causes background to be measured only for the extended energy range 15 2000 keV Total background CPM is calculated for all detectors that are in use In addition for each normalized isotope the background CPM in the counting window of the isotope is determined If background normalization is done in 1470 using the conveyor there may be detectors that are active but nevertheless are not used for measurement since with the conveyor only active sets of adjacent 10 5 2 or 1 detectors can be used for measurement 10 1 2 Total background CPM This is calculated by summing counts from all multi channel analyser MCA channels except the first 10 This channel range is called the open window Since the last MCA channel includes high ene
219. sults are handled These depend on the type of communication protocol you have The normal situation is as follows Buffering of results is selected the SYSTEM parameter Printout selections Without buffering to PC is NO If counting is started by pressing the instrument START key or from MultiCalc by pressing FI COUNTER and selecting WIZARD measurement results are buffered in the instrument MultiCalc will evaluate the final results and print these out as determined in the counting protocol Buffering is not selected SYSTEM Printout selections Without buffering to PC is YES If counting is started from MultiCalc or by pressing the instrument START key measurement results are sent directly to MultiCalc without buffering This option is recommended only with the Resident filer installed which case you should have the communication protocol WIZARD BG instead of the normal WIZARD The following special cases are also possible 1 SYSTEM Operation mode Evaluation is RiaCalc WIZ or Cpm and SYSTEM Printout selections Without buffering to PC is YES In this case measurement results are sent directly to MultiCalc without buffering 2 SYSTEM Operation mode Evaluation is RiaCalc WIZ or Cpm and SYSTEM Printout selections Without buffering to PC is NO In this case measurement results are not sent at all to MultiCalc 5 6 The FILES function Most FILES functions are not available in CPM mode they
220. t onto the conveyor Insert an empty rack after the last normalization rack to stop the conveyor When you have loaded your normalization rack s and stop rack press START Repeat this normalization procedure for each isotope to be used Wait until the complete report for each isotope has been printed out Make sure that the efficiency is within the limits 0 9 1 1 Make sure you do not leave a background normalization or isotope normalization rack on the conveyor when you have finished with it otherwise you may start an unwanted new normalization and lose the previous results 6 1 4 Printout columns The printout column headings are DET PEAK CHN PEAK DEV RESOL ACTIVITY COUNTS EFFICIENCY ERROR where ERROR 100 Counts WINDOW keV LOW HIGH DECAYED ACTIVITY MEASURED COUNTS DETECTOR EFFICIENCY RELATIVE ERROR HORROCKS EFFICIENCY STANDARD CPM SIGNIF LEVEL these only appear if extended normalization printout has been selected these two items do not appear for single detector instruments If the instrument has several detectors installed but only one of them is active these fields are printed however Then efficiency is 1 0 and the error 0 0 See also section 10 2 17 6 1 5 Normalization sequence No special dual label normalization sequence is necessary for the following reasons During normalization each time an isotope is counted with one detector the spectrum obtained from each detecto
221. t results to be output to a printer connected to the parallel printer port 7 3 6 4 Without buffering to PC There is a result buffer on the hard disk It enables you to disconnect the PC at any time without interrupting counting It is also possible to start the counting from the keyboard of WIZARD by pressing the START key and save all assay printouts to the buffer and later connect the PC and evaluate the results If there is not enough space on the instrument hard disk the program stops If 999 assays have already been saved in the buffer then WIZARD starts writing over the oldest buffer files You should evaluate the old assays from the buffer well before the hard disk becomes full or there are 999 unevaluated assays in the buffer The size of the instrument hard disk is about 30MBytes The assay results in the buffer can also be deleted without sending them to the PC If you are using MultiCalc then results are buffered when this parameter is NO if counting is started by pressing the instrument START key or from MultiCalc with the command B S when the device parameter PC is not used Results are sent directly to MultiCalc when this parameter is YES If you are using the MultiCalc resident filer set this parameter to YES If the RiaCalc WIZ or Cpm modes are used results are never buffered In this case if this parameter is YES results are sent to PC otherwise not 7 3 6 5 Write results to file In addition to sending results to the
222. t time The exact time to the nearest 1 10 second when measurement of each sample was started can be included in CPM printouts If this starting time output has been selected a field named CLOCK appears at the right end of the CPM printout The measurement start time field can be enabled and disabled in the following way Set the SYSTEM parameter Diagnostic output Print meas start time to Yes to enable the field and to No to disable it 33 CPM operation 3 6 3 Run ID Each time a batch of samples is run it is given a run ID This will be printed at the beginning of the CPM results The run ID is specific for each protocol This enables you to distinguish multiple runs of the samples with the same protocol ID 3 6 4 Dead time factor The Dead time factor can be included in the CPM printout To do this set the SYSTEM parameter Diagnostic output Print dead time factor to Yes Dead time factor is explained in 10 1 3 3 6 5 Bad spectrum results Note the output values for counts and CPM are 0 if the spectrum is bad e g if the coincidence peak is missing or too small for I 125 3 7 Leaving the editor Press EXIT to leave the editor A choice of three possibilities is shown Save changes and exit Saves the parameter setting on file and leaves the editor Quit and ignore changes Leaves the editor without saving the changes Edit You return to the editor to do further editing 3 8 Running an assay Make sure that
223. tained in GLP test normalization There are four options some of which lead to other options View allows you to view the GLP data Delete delete GLP data Criteria allows you to select the warning limits for different types of GLP data You can select Isotope you can select the isotope type from those for which a GLP normalization has been done Item many items appear from which you can select the one which you want the GLP data PEAK BGRD EFFIC RESOL EFFICIENCY COVERAGE CHI PROB WIN CPM TOTAL CPM Print criteria select the isotope for which you want the criteria to be printed out 4 9 11 Waste log file A waste log file that contains the total CPS and DPS values of all measured assays isotope normalisations and GLP TEST measurements can be printed or stored on a datalogger disk The file can also be deleted or sent via the PC port to an external computer The waste log can contain approximately 700 entries after this the older half of the entries is deleted and the log starts growing again For isotope normalization and GLP test TOTAL CPS is the average corrected sample activity in the isotope counting window in all detectors used It is corrected for dead time background activity and isotope decay TOTAL DPS is the average corrected sample activity in the open window divided by Efficiency which is a SYSTEM parameter that appears after you select Isotope and then the isotope name For assay measurement TOTAL CPS is t
224. te_keV 30 367 1 045872 ELSE Detector_keV Absolute_keV END Detector_keV Detector_keV MCAoffsetForTheUsedEnergyRange MCA offset is set with the 1480 service program separately for both energy ranges For the 1470 counter model it is always zero The function to convert from detector keV s to absolute keV s is the inverse of the one above As was said before the effective gain value during isotope normalization using a fixed window is equal to the nominal gain 10 2 4 2 Dynamic keV and Dynamic window These window types take into account that the effective gain is not always exactly the same as the nominal gain but may vary according to e g temperature and measured activity The effective gain is calculated by dividing the main peak energy in detector keV s by its observed MCA channel number To do this the program must know the peak energy and be able to find the actual channel number of the peak The peak energy is given by the parameter SYSTEM Isotope lt isotope name gt Peak pos keV The next sections explain how the peak channel number 1s determined 10 2 4 2 1 Finding the isotope main peak Finding peaks in a spectrum is a pattern recognition problem The analogue to digital A D converter is not ideal so in reality the MCA channels are not all equally wide This together with the statistical nature of isotope decay makes the spectrum ragged In order to recognise a peak we must have some idea where to seek 1t an
225. tector block and slide the tubes protruding from the bottom of the tray into the detector wells as shown in the figure 7 2 5 Counting samples Press the START key The display will show Measure manual mode Select the type of measurement or press STOP to cancel measurement Measure an assay GLP test Measure background Normalize detectors Press START to measure Select Measure an assay and press E A list of available protocols will appear Select the one you want and press E The display will then show Ready to measure batch number 1 To start insert samples and press START key To end press STOP key When you have loaded the sample tray press START Counting will start in the normal way and results will be displayed and output When the tray has been counted you will be requested to load the next tray with the following message Ready to measure batch number 2 To start insert samples and press START key To end press STOP key When the last batch has been counted press the STOP key 7 2 6 Returning to automatic counting Remove the sample tray from the detector block and close the cover Then go to System and select Operation mode and then change the Use manual mode to NO The elevator arm will return to its normal position and WIZARD will be ready for automatic operation Note In Manual mode the counting can be started from MultiCalc only in offline mode not in online mode 90 7 2 Manual operation
226. tectors not in use 6 7 H 9 10 DECAYED MEASURED DETECTOR RELATIVE SIGNIF DET ACTIVITY COUNTS EFFICIENCY ERROR LEVEL 1 1 0000 68182 1 0138 0 31 78 252 2 1 0000 68475 1 0182 0 30 0 279 3 1 0000 65925 0 9804 0 31 68 787 4 1 0000 65654 0 9763 0 31 35 961 5 1 0000 68010 1 0113 0 31 20 969 AVG 67249 1 0000 0 31 VALUES SAVED END OF ISOTOPE NORMALIZATION Figure 11 Diagnostic info printout in conjunction with an isotope normalization Detector 6 is not active SYSTEM Isotopes I 129 Counting window is Fixed NORMALIZATION OF 4 I 129 26 Aug 1994 10 33 07 DIAGNOSTIC INFO FOR DETECTOR 1 Dead time factor 1 01552 Open window counts 35789 Spectrum assessment Spectrum OK ISOTOPE I 129 Expected peak chn 31 0 Low channel limit 20 High channel limit 46 Used peak channel 32 2 Peak begin channel 13 Peak end channel 56 Actual coverage 98 51 Window shifted Peaks in smoothed spectrum that exceed 576 Channel Counts 32 2867 END OF DIAGNOSTIC INFO Total counting time 15 Detectors not in use 6 7 8 9 10 DECAYED MEASURED DETECTOR RELATIVE Resol start chn 26 Resol end chn 38 DET ACTIVITY COUNTS EFFICIENCY ERROR 1 1 0000 35254 0 9955 0 43 2 1 0000 35478 1 0019 0 42 3 1 0000 35377 0 9991 0 43 4 1 0000 35356 0 9984 0 43 5 1 0000 35585 1 0050 0 42 VALUES SAVED END OF ISOTOPE NORMALIZATION Figure 12 Diagnostic info printout in conjunction with isotope normalization Detector 6 is not active SYSTEM
227. the ZT extension to make more space on the disk Error messages concerning the datalogger DATA DISK NOT READY 1 RETRY 2 0MIT This could happen if no data disk was in place Check this before selecting Retry DATA DISK ERROR 1 RETRY This probably means that there is a fault on the disk itself or the write protect switch was on Either change the disk or move the write protect switch as appropriate DATA DISK FULL In this case you must either change the disk or delete some of the data saved on it Notes CPM output see section 3 6 for information about CPM output This information is also valid for the CPM output to the datalogger Datalogger file name printed the name of the corresponding datalogger file is printed after each measurement for which data is saved in a file Run ID number is included also in the CPM output sent to the datalogger This can be used to link printed assay results to stored assay spectra files 7 3 6 6 Horizontal background printout Background CPM values for different isotopes can either be printed vertically or horizontally Specify the layout you want 7 3 6 7 Printer type The following two printer command languages ar supported Epson FX and HP PCL3 Epson FX Most dot matrix printers support this HP PCL3 Many Hewlett Packard printers support this 7 3 6 8 Printout selections Print page numbers If Yes a header PAGE n wwhere n is the page number is printed on to
228. the selected operation is Create then the line Counting time will appear in the file operation submenu In principle each measurement result in a data input file has its own measurement time However in practice the measurement times for all samples in an assay are often the same When a new data file is created the counting time given here is assigned to all measurements in that file This makes the creation of data files easier Edit An input data file consists of a series of records one for each stored measurement result Each record contains the measurement time measured counts CPM s and their errors for one or two channels 54 4 Operation with internal RiaCalc WIZ This editor shows only the CPM values and the measurement time of the first measurement in the file When the data file is saved this measurement time is assigned to all records in the file and for each record the counts and error fields are calculated based on the measurement time and the CPM value s You can move up or down one page by pressing the PgUp or PgDn keys respectively and to the beginning or end of the file by pressing the Home or End keys respectively To edit a measurement or to jump to a specified line number press the E key If you have created a new data file and have not added any CPM s to it yet try the following shortcut Use only the E key to move from one menu or menu item to the next appropriate one when you enter new CPM values The menu items wh
229. this problem for WIZARD 8 13 Isotope selection WIZARD has a list of isotopes see the list in the next chapter 8 2 Specifications All the isotopes in WIZARD have predefined settings The energy range of samples which WIZARD is set to cover is up to 1000 keV which means that WIZARD can handle Chromium Release tests as well as the usual single and dual labelled RIA s The settings for these isotopes can be customized by the user in the range up to 1000 keV The energy range can be extended by qualified service personnel 8 14 Multichannel analyzer There is a linear multichannel analyzer with 1024 channels calibrated for the range 1 1024 keV It has a 12 bit Analogue Digital converter and automatic dead time compensation 8 15 Automatic and manual operation WIZARD is both an automatic and a manual counter Manual counting allows you to use even bigger samples than in automatic mode It also gives you security in case you should have problems with the conveyor you can still count samples manually If you just want to count a few samples without bothering to put them into a rack and load it onto the conveyor you can do it with manual mode 8 16 GLP performance testing Instrument performance can be monitored by running GLP TEST normalizations at regular intervals These store data that can later be viewed in graphical format GLP means Good Good Laboratory Practice gt Laboratory Practice A GLP TEST normalisation is similar to i
230. time in seconds CONC concentration value This is printed as follows SEO PAT TIME CONC PERE PERE PERRE EERE HHEH where the mark is used here to define the field length Default field lengths are listed in section 4 10 Assume now that you want the SEQ printout field to start from position 15 and that there should be 5 spaces between TIME and CONC Proceed as follows Select sample result output Then select lt Left margin of paper or display gt and press E Select Make new item after this one and press E two times Select MOVE TO A menu will appear on the screen Change item type MOVE TO Change item value 10 Use the numeric part of the keyboard to change the item value to 15 Press EXIT You will see a new print selection 51 4 Operation with internal RiaCalc WIZ lt Left margin of paper or display gt 15 lt move to this column position gt SEO sequence or tube number Select TIME counting time in seconds and press E Select Make new item after this one and press E two times Select MOVE RIGHT move to the right and press EXIT A menu will appear on the screen Change item type MOVE RIGHT Change item value 10 Use the numeric part of the keyboard to change the item value to 5 then press EXIT 4 6 9 Printout switches This selection offers you further choices to modify the printout format In its initial setting both individual result
231. tion parameters you must set the baud rate number of data and stop bits parity and handshake signal for each port Select the appropriate port the choices are PC port PRINTER port Select the appropriate parameter Baud rate 300 1200 1800 2400 4800 9600 Stop bits 1 2 bit s Word length 7 8 bits Parity None Even Odd Protocol None DTR RTS XONXOFF The default items are underlined 7 3 10 3 Output device status All output devices can be switched on or off If all devices are off no results are output but the counting continues normally The user can override these settings from the PC and when on line counting is started from the PC the CPM results input data to an evaluation program are sent from the PC port to the PC in any case During counting the instrument program checks if the output device used is connected break signal and ready off line on line signal If not an error message is seen on the display At this point the user can correct the error and retry or omit the device No characters are lost in the first case and in the second case the device is omitted until the counting is stopped and another one started The same applies to the case when the output device is busy ready busy signal and the instrument cannot send data out 7 3 10 4 Error messages The following error messages concerning output devices may appear on the built in display PRINTER not connected l retry 2 omit PRINTER not ready
232. tions You can now replace the back panel and front cover PAZ 11 5 Checking the mains voltage setting Measure and note the mains voltage at the outlets to be used Locate the mains selector switch see the figure below this is on the left side when looking from the rear of the instrument If necessary adjust the mains selector switch to correspond with the measured supply Check that the fuses fitted in the fuse carriers are of the correct rating for the local supply and according to the label POWER The mains selector in older instruments has the 200 VA 50 60Hz MAINS voltage marked on it and is combined with the SV AGH fu iers To adjust the mains selector pull 230V ACH15V AC fuse carriers To adjust the mains selector pu It out turn it through 180 degrees and replace Fuses 115V AC T4A H 250V it b Fuses 230V AC T2A H 250V It FUSE S MAINS INLET W AOZL 0LL 220 240v 11 6 Connecting up the counter and peripherals Connect the counter to the PC if supplied and the printer using the cables shown in the Peripheral Installation Sheet Further details of the settings for the PC and printer are given in their respective installation sheets which follow these installation instructions WIZARD has three communication ports They are 184 11 Installation 1 PRINTER SERIAL RS 232 port to connect a se
233. tiveCpmError RelativeError1 RelativeErrorInRelativeDetectorEfficiency Here RelativeErrorInRelativeDetectorEfficiency 0 if the counter has only one detector in use CorrectedCpm1 is corrected for dead time and background but not for relative detector efficiency spillover crosstalk or isotope decay Note that if the final corrected CPM in the RIA IRMA RATIO CPM printout includes spillover or crosstalk corrections the ERROR printed still refers to the CPM value which has only dead time background and relative detector efficiency corrections but not spillover or crosstalk corrections In this case it is not possible to also output that CPM value to which ERROR refers 10 3 8 Values that are printed in diagnostic info Diagnostic info is used to show some measurement results that are not included in background or isotope normalization printouts or assay printouts Diagnostic info is printed if SYSTEM Diagnostic output Print diagnostic info is Yes For 1470 you can select the detectors for which diagnostic info is printed in the menu SYSTEM Diagnostic output Select detectors for info The parameter SYSTEM Diagnostic output Level of diag info is used to select how much data is printed Below are two examples of assay diagnostic info printout 164 10 3 RIA IRMA RATIO assay counting ASSAY 26 Aug 1994 10 38 01 Protocol id 29 PROTO1 Time limit 20 Count limit 99999999 Isotope I 129 Protocol date 22 Aug 1994 10 59 52
234. tom of the screen there is a status line This shows what is being measured In the display example above the status line is Press START to measure The status line can have the following texts Press START to measure Measuring an assay Measuring background Normalizing Clearing the conveyor Seeking assay GLP test 2 4 Live display When samples are being counted the display can show actual counting values either collected counts collected counts per minute CPM values or a complete isotope spectrum The word Live indicates that the display is working in real time values are updated at the same pace as counts are accumulated 2 4 1 How to use the Live display In order that the Live display works the counting must be actually happening Select the OPERATE menu The status line must show Measuring assay or a corresponding text which indicates that counting is active Select Show cpm results and press the E key Counting parameters are shown see the figure below 2 4 2 Display modes There are five display modes Note there are two counting windows in dual channel counting Show cpm results Measuring now elapsed time is Counting parameters Measurement Protocol 11 PROTO3 Label I 125 Preset time 60 Counts limit 9999999 Batch number 2 Change data with keys 19 2 Wizard controls Counting parameters as shown above COUNTS Accumulated counts in counting window CPM Counts per minute i
235. trum of the repeat measurements 81 7 Additional WIZARD functions 7 1 Dual label counting 7 2 Manual operation 7 3 System mode 7 4 STAT counting 7 5 Power failure 7 6 Routine maintenance 7 7 Safety information 7 8 Large Eppendorf tubes 83 7 1 Dual label counting 7 Additional WIZARD functions 7 1 Dual label counting 7 1 1 Introduction Dual labelled samples have actually two independent analytes in the same vial In order to separate the radioactive labels from each other the labels must have separate energies The most common dual label assay in practical work is the B12 Folate assay in which labels are Co 57 and I 125 Before running a dual label assay you must normalize the instrument for both isotopes as described in Part 6 The analysis for dual labelled assays requires two protocols to be set one for each isotope used The two protocols are connected by specifying in protocol A the name of protocol B Protocol A is a master protocol and B is the slave Channel A protocol Channel B protocol Refers to channel B protocol Figure showing relationship between protocols in dual label The program uses the word channel to describe the counting process controlled by a particular protocol Channel A counting is controlled by protocol A and channel B by protocol B Each channel may include counts from both labels so the dual label program has to disentangle the counts so as to arrive at a pure counts value for
236. ultiCalc In order for the printing to succeed MultiCalc must be receiving data from WIZARD and in the WIZARD communication protocol the Terminal parameter must be VT 52 and in the MultiCalc system parameters the Printer parameter must be set to Epson FX 10 2 16 How the relative error of relative detector efficiency is calculated In normalization printout this quantity has the title RELATIVE ERROR The following formulae are used Variance1 SQUARE DeadTimeFactor 60 CountingTimelnSeconds CountsInWindow RelativeErrorlnBackgroundCpm 1 0 SQUAREROOT BackgroundCountsInWindow Variance2 Variance1 SQUARE BackgroundCpminWindow RelativeErrorInBackgroundCpm Variance3 Variance2 SQUARE DecayCorrectionFactor Let CorrectedCpm be the CPM value in the counting window that is corrected for dead time background activity and isotope decay Then the relative error of CorrectedCpm is RelativeErrorOfCorrectedCpm SQUAREROOT Variance3 CorrectedCpm 150 10 2 Isotope normalization and GLP TEST measurement Let AverageCpm be the average of CorrectedCpm s for all used detectors Then for each detector the relative efficiency is equal to its CorrectedCpm divided by AverageCpm If the counter has only one detector in use then AverageCpm CorrectedCpm and from this it follows that in this case RelativeDetectorEfficiency 1 and RelativeErrorInRelativeDetectorEfficiency 0 Otherwise the following formulas are used to calculate Relativ
237. um 12 2 2 2 30 9985 19766 9 1 74 35 20 9 66 28 13 2 3 2 30 9883 19732 5 1 74 30 8 8 149 36 14 2 4 2 30 9962 19672 5 1 75 29 19 1 65 67 15 2 5 2 30 8502 17072 4 1 82 25 4 5 266 89 16 2 6 2 30 8550 17143 5 1 82 28 2 7 483 22 17 2 7 2 30 7086 14172 5 1 92 24 11 5 107 39 18 2 8 2 30 6920 13851 2 1 94 32 6 8 198 86 19 2 9 2 30 19139 38353 0 1 46 22 2 7 424 70 20 2 10 2 30 21855 44981 5 1 41 27 20 5 105 55 END OF ASSAY Figure 9 A dual label RIA IRMA RATIO assay printout when SYSTEM Operation mode Evaluation is Cpm In position 11 there was an l 125 tube 10 3 1 2 1 Neither isotope has crosstalk correction This means the for both isotopes SYSTEM Isotopes lt isotope name gt Crosstalk correction is No The spillover correction factors are calculated in the following way During isotope normalization the spectrum of the normalization sample is stored for each detector in the instrument hard disk Latest background spectra are also copied and stored with normalization spectra The program determines counting windows in terms of MCA channels This is done by converting the stored keV values of counting window boundaries by using the effective gain during the isotope normalization that produced the spectrum on which the counting window is used If SYSTEM Isotopes lt isotope name gt Counting window is Dynamic the window boundary keV values are determined during isotope normalization on the basis of the parameters
238. ummed in this spectrum over the counting 147 10 2 Isotope normalization and GLP TEST measurement window for detector L this is then corrected for dead time converted to CPM background activity is subtracted and the result is corrected for the relative detector efficiency for detector L Finally the value is decay corrected with the decay correction factor calculated for the batch having the normalization sample in detector K if decay correction has been selected The following NxN matrix is set up where N is the number of detectors that are in use CPM in detector 1 using the counting window of detector 1 when the normalization sample is in detector 1 CPM in detector 2 using the counting window of detector 2 when the normalization sample is in detector 1 CPM in the last used detector using the counting window of the last used detector when the normalization sample is in detector 1 CPM in detector 1 using the counting window of detector 1 when the normalization sample is in detector 2 CPM in detector 2 using the counting window of detector 2 when the normalization sample is in detector 2 CPM in the last used detector using the counting window of the last used detector when the normalization sample is in detector 2 CPM in detector 1 using the counting window of detector 1 when the normalization sample is in the last used detector CPM in detector 2 using the counting window of detector 2 when the norma
239. unknowns aX dX t AyxXy D Ay Xk tay t tayyXy by as the matrix equation au Ay Xi b eee eee eee eee eee Ay Any Xy by If we denote GQ Ay Ay Any x K Xy 177 10 5 Appendix Some basic mathematical formulas we can write the equation as The inverse of a matrix Mis denoted with A and is defined so that K MB for all K and B that fulfill the previous equation It is possible to find an inverse for every NXN matrix for which the determinant is not zero Matrix inversion and multiplication can be used to solve a system of N linear equations each having N unknowns In this Calculation Methods manual the following situations are modelled with a system of linear equations e A sample consists of several say N isotopes and as many counting windows are used when it is measured Each isotope is assigned one counting window When pure isotope samples are measured the spillover factors giving the relative activity of an isotope in the counting windows of other isotopes as compared to the isotope s own window can be determined and an NXN matrix can be set up as descibed in Section 10 3 1 2 1 on page 158 and Section 10 4 1 on page 171 The spillover factors can be assumed to independent of isotope activity at least as long as the isotope activities are not very large When an unknown sample containing several isotopes is measured the inverse of the spillover factor
240. ve gain is calculated as described in the previous section and then the window boundaries are converted to MCA channel numbers using this gain value 10 2 4 2 3 Dynamic window in MCA channels If SYSTEM Isotope lt isotope name gt Counting window is Dynamic the counting window is determined by the parameter SYSTEM Isotope lt isotope name gt Window coverage The window is set around the observed isotope peak so that it is as small as possible but still contains at least the preset coverage fraction of all counts in the open window Once this window has been determined its boundary channel numbers are converted to keV s and stored with isotope normalization results If SYSTEM Isotope lt isotope name gt Spectrum type is I 125 the counting window always includes the area between the isotope main peak and coincidence peak 141 10 2 Isotope normalization and GLP TEST measurement Background activity is not taken into account when window coverage is calculated 10 2 4 2 4 Calculation of Horrocks efficiency for I 125 Horrocks efficiency is calculated when SYSTEM Isotopes lt isotope name gt Spectrum type is I 125 and SYSTEM Isotopes lt isotope name gt Counting window is Dynamic or Dynamic keV In this case in extended isotope normalization printout the column title HORROCKS EFFICIENCY appears Horrocks efficiency is not calculated if Counting window is Fixed Horrocks efficie
241. w coverage For window types Dynamic keV and Fixed the boundaries are given by the isotope parameters SYSTEM Isotopes lt isotope name gt Low boundary keV and High boundary keV If Counting window is Dynamic or Dynamic keV then the effective gain is calculated by dividing the energy of the main isotope peak in keV s by the MCA channel number of the main peak If Counting window is Fixed then the nominal gain is used Next the activities of all standards in all counting windows are determined This is done in the following way For each isotope standard spectrum counts are summed over all isotope counting windows They are corrected for dead time converted to CPM and the background activity is subtracted Dead time factor background activity and possible isotope decay correction factor are all determined by the isotope spectrum used The effective gain which is used to convert counting windows from keV s to MCA channels is taken from the isotope normalization which produced the counting window and not by the isotope spectrum to which the counting window is applied In RIA IRMA RATIO dual label assays the gain is determined by the spectrum on which a counting window is applied and theoretically it should be this way But since isotope normalizations usually are done at the beginning of a MIA so that the effective gain has no time to change and because the detector unlinearity correction described in Section 10 2 4 1 2
242. weak sample is in the first batch of an assay then either the same counting window is used as was used in the most recent assay run with that isotope or else this is determined by the parameter SYSTEM Operation mode Default is norm window Window coverage Determines how wide the counting window is set in isotope normalization It is the fraction of the counts of this isotope in the whole spectrum that must fall within its counting window The window is first selected to be as small as possible but still to contain at least the specified fraction of counts Then it is further widened five channels in both directions Low boundary keV High boundary keV These are the boundaries of the counting window The unit is keV Although the editor permits you to give values up to 6000 keV in practice the hardware imposes a limit which is given by the menu line MCA high limit Note If you want to measure high energy photons set the counting window to include the last MCA channel This is because the last MCA channel collects all photons which have a higher energy than the high limit of the MCA For example to measure all photons registred by a detector set the Counting window to Fixed the Low boundary keV to 0 and the High boundary keV to e g 3000 For 1470 the MCA high limit is about 1024 keV assuming the gain set at the factory has not been changed Since the High boundary setting exceeds this value the last MCA channel is inclu
243. wn Save changes and exit Saves the parameter setting on file and leaves the editor Quit and ignore changes Leaves the editor without saving the changes Edit You return to the editor to do further editing 4 8 Running an assay Make sure that the first cassette has 1ts correct protocol selection ID and that the appropriate protocol is stored in the instrument For more information about the ID system see section 2 5 Put a STOP ID on the last sample rack to be counted or use a STOP rack or a totally empty rack in order to stop the instrument automatically Load racks starting with the right hand conveyor lane Start counting by pressing the START key The instrument will now count all the loaded samples and the RiaCalc WIZ software will evaluate the final results and output them as determined in the counting protocol During counting you can see the live results on the live display as described in section 2 4 Alternatively you can select the menu item Show evaluation results to see the output from RiaCalc WIZ on the display The counting will stop automatically when a STOP rack is found You can also press the STOP key on the WIZARD keyboard In that case the following text will appear Continue End assay continue End assay Clear conveyor Depending on whether you want the next assay to be counted or all counting to stop select End assay continue or End assay clear conveyor respectively To override the stop instru
244. ype REFER Save replicate array Yes Cut this item Paste item after this one Make new item after this one Choice use gt or ENTER In the example REFER was the item selected to be edited If you want to change the coding item type select it and press E You will then get a list of all possible item types Select the one you want and press EXIT twice 46 4 Operation with internal RiaCalc WIZ Edit protocol 11 PROTO3 Edit item New item type REFER response when conc 0 TOTAL total labelled antigen BLANK non specific binding POS positive dose limit NEG Negative dose limit STD Standard dose sample UNKN unknown sample and dilution REPEAT remainder of samples n times Select and press ENTER The main control list will then show the new selection you have made The recommended order for sample tubes is blanks NSB totals references zero sample standard samples unknowns Unknowns are either patient samples or control samples in arbitrary order the controls must be further specified on the line called Controls There is also a parameter Repeat remainder of samples n times where n can have the values 1 to 32767 or infinite This allows you to repeat count those samples which follow the point in the list at which this parameter is placed 4 5 16 Options Unkn CV flag limit A flag can be set for unknowns by typing the CV of the concentration abov
245. zation nere 119 Customized output sess 48 D Datalogger iiiter lia 98 Error messages 100 A rre intere ri t sees 93 A eerie iste er Het iei 127 Dead time correction ooooccnccocnnonocnnoncnoncnononnconoco nono nonnono 131 Dead time factor nennen dni 34 DA nennen 94 Decontamination ooooonoccnononcnooncnnonnnonccn nono nonncnnc crono nme 111 Delete protocol seen 31 41 Detector nuQ ETE P S Detector inactivation Detector number and normalization Detector Usage reor rrr rer irre eere pets Ibzcg eC AR Matching Dets only at odd positions sss Device status output Diagnostic output Disable dead time correction asserens sisi 104 Read spectra from diskette Dilution series averages cnnsneneenennennneenennennn nn Dimensions hr A Disable background correction sss 104 Disable dead time correction sess 104 Display modes ENS displasia a aia Live display valueg 5 caa aiii ca Live spectrum display Dual evaluation esses Dual labels id eia 13 Index Constraints on protocol setting neeneeen 86 Parallel evaluation cnc Successive evaluation da Dual label counting seen dual label selection Dynamic normalization parameter Dynamic keV i scissisecescscas casts ciere tpe poni os ena sacas 95 E

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