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Anthos HT-series Plate Reader operating manual

1. ceeeeeee eee ee teens nn en nn 43 6 1 1 TO COPY altesti n n DR er nu Penn 43 6 2 Print Test Printout of all programmed test parameters unensennennnnnnnnennennn nn 43 6 3 Dir Printout of the entire test directory zsensenseneeneenenn none nn nun nun nun nun nennen nennen 44 6 4 Clear Erasing individual tests or the entire test Memory uneannnnennennnnnnn nennen 44 7 RUN TEST MEASURING A PLATE WITH A PREPROGRAMMED TEST 45 7 1 Me suremMent Resultsi n n ea er en a aha hir 46 7 2 Plate Layout ar nenne 47 7 3 Qu ntitativerdisplays tra teenie seele 47 7 3 1 Standard CUVE seision i eaa ea DDE nella 47 7 3 2 Standard Dita since ni ieeiuiseineete 48 7 4 Quantitative Results uussnsennnnnnnnonnonnennennnn nn nn nun nun nun nun nun nun nun nun nun nun nun nun nenne 49 7 5 Qualitative Resultszr asien ee 49 7 6 Kinetie results displaysna eenree nie nie 49 7 6 1 Kinetic Raw Data sn a ne Ann eh Da er deeds De een 49 7 6 2 Graphit a a ha a nn na a EAA TEA EAA 50 7 6 3 Zoomed Graphics isisisi aa gees an na He a ae nnd 50 7 7 Relational Results enaa dcteea veg ides ne en ehe nn ren 50 8 PLATES PLATE MEMORY RE EVALUATION OF PLATES WITHOUT MEASURING PRINTOUT OR VIEW OF MEASUREMENT AND EVALUATION DATA ee ra nen nee 51 9 JERROR MESSAGES ann a Ra A A A 53 9 1 Error messages in the display s sssssssssesssnrsrrssssrrrrrnessnrrnrnnensrrrnnnsenerrnnnnns 53 9 2 Error messages and w
2. SAMPLE ID POS MEAS MEAS_STAT CONC CONC_STAT GROUP GROUP_STAT RELAT RELAT_STAT The data fields can contain the sample number ID the plate position POS the measurement value MEAS the status of the measurement value MEAS_STAT the concentration CONC the concentration status CONC_STAT the group GROUP the group status GROUP_STAT the relation values RELAT the status of the relation values RELAT_STAT 10 6 4 Blanks record format Display of validity range and positioning Blank PLATE COLx ROWx POS MEAS MEAS_STAT The data fields contain the blank s validity range operating manual 72 anthos ht1 ht2 ht3 e on the entire plate PLATE e for one column COL 1 12 e for one row ROW 1 8 In addition they contain the blank s position on the plate POS its measurement value MEAS and the measurement value status MEAS_ STAT 10 6 5 Control record format Display of the set controls The line shown below is wrapped Control ID POS MEAS MEAS_STAT CONC CONC_STAT GROUP GROUP_STAT RELAT RELAT_STAT The data fields can contain the sample number ID the plate position POS the measurement value MEAS the status of the measurement value MEAS STAT the concentration CONC the concentration status CONC_STAT the group GROUP the group status GROUP_STATUS the relation value RELAT the status of the relation value RELAT_STAT 10 6 6 Cutoff record format Display of the set
3. 4 By having programmed Outputchannel RS232 in the Reader menu Setup Tests and having performed a measurement using this test Note Transmission of pure ASCII data as per result data string pattern e The data are merely separated from each other by space The entire data file is divided into individual blocks or fields e Only those blocks needed for the respective test are transmitted e g no quantitative data for a qualitative test e The block length depends on the complexity of the test used e Blocks are unmistakably identified by their headers e Each block begins with a character and a key word e g header representing the block beginning e It ends with the character for the beginning of the next block or the end mark End at the end of the data file e The following key words have been defined Header Message Sample Blank Control Cutoff Standard Curve Basically two different types of blocks have been defined 10 5 1 Blocks with multiple information Example Header Format 110 They consist of one or more lines each line able to contain numerous information units Only blocks with the key words Header Message or Curve at the block beginning can contain this type of information block beginning character Header key word Format description of information 110 information Plate ID 12234781 Data line 10 5 2 Blocks with the same type of information record
4. 3 0 OD Change to extended range indication range 0 3 5 OD by pressing CE 25 anthos ht1 ht2 ht3 Note in Extended Mode the indication limit is extended to 3 50D Measurement time is doubled and the use of wavelengths under 400 nm are not permitted Turn page with F5 NEXT 5 3 Premeasurement functions optional parameters for functions being performed prior to measurement FRE SHAKING Duration Intensity PRE WAITING MOHITORING i KERI Well Positions Bal Cal Monitoring Mode ABSOLUTE D lower limit ZEa_m LC D D urrer limit 1664 mob Read Interyal 5 sec select item cursor modify CE A an A A E e Preshaking Plate Shaking to improve the fluid homogeneity at a maximum filling capacity of 200 ul per well Change to YES by pressing CE enter duration in seconds using the keyboard and select shaking intensity from three levels low medium high e Prewaiting Waiting time prior to measuring Set to YES by pressing CE enter desired time in seconds using the keyboard Maximum time 99 sec e Monitoring Automatic observation of certain wells Measurement is started upon reaching a certain limiting value Change to YES with CE Additional settings e Well positions Setting the plate positions to be observed Press CE position cursor in the menu select with F4 SELECT delete with F3 CLEAR or F2 CLEAR ALL confirm with ENTER The selected positions may be observed in
5. can however also be defined by an algebraic formula containing all the set controls standards and numerical constants Note When entering the formula make sure that the sequence for the response values is either ascending or descending Press CE move cursor to the row in the display menu with the desired character and confirm with the corresponding function key 5 6 2 How to store a standard curve Note The use of stored standard curves is not generally recommended or applicable for the quantitative evaluation of ELISA s It is explicitly the responsibility of the user to take advantage of the described software possibilities in the appropriate way Once a standard curve has been measured and OD results have been obtained by programming for standard S1 the response value as Si for the standard S2 as S2 etc the curve can be copied in the following way Make a printout of the measured test containing both concentration values and OD values Begin definitions of a new test The layout need not contain a real standard anymore As response values enter the OD values of the standards from the printout protocolof the measured test Now a new plate can be measured that will be evaluated by means of the stored standard curve To be able to correct possible deviations of the curve it is highly recommended to have at least one real standard on the plate by which a correction factor can be calculated Example A plate with 5
6. 1 low 2 medium 3 high shaking duration maximum 60 sec end of text The Reader acknowledges the command executes the shaking command but does not send an answer string back 10 4 13 B Transmission of plate identification number via barcode Command string to the Reader STX B ETX start of text key B read barcode label end of text The plate carriage enters the device and an attempt is made to read the label via plate barcode Upon acknowledgement the plate identification number is sent as an answer string to the computer Example STX 100001 Cxx ETX 10 4 14 R Repeat repeat of the last data transmission If the last data transmission from the device was faulty e g checksum error it can be repeated using the command R Command string STX R ETX 10 4 15 Error messages operating manual If the measuring process is interrupted light energy fluctuations transport system stop key communication is discontinued and an error message containing an error code number and a short description of the type of error is sent to the computer 69 anthos ht1 ht2 ht3 Example STX E 01 Plate transport Cxx ETX 10 5 Transmission of evaluation results 1 By selecting the output channel RS232 in the Reader menu Plate or Run Test 2 By entering the remote control command TR Run Test PR Plate Results 3 By selecting Transmit with the function keys in the Reader menu Plate Run Test
7. 1 4 3 POWER SUPPI ree innen a aaa a en od cand nenn he RD er tee nn Dee 10 1 4 4 Datla connection 542 ee ann ee ee aeg 10 1 4 5 Automato hesena anaE a EREE Eaa aa DE EEDI pal a aE N ENE aea EDA 11 1 4 6 Supplementary equipment zeusssensnennnnennnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnennnnnnnnne 11 1 57 Cleaning Hr Dee Eee 11 1 6 PROCOMEY settee el ne pa ehe 11 1 7 Scope Of SUPPIY rn a aan ann 12 1 8 Warning and directions u 2 2 en 12 1 8 1 Environmental operating conditions 4 4 r rannnnnnn nennen nn nn eens nets ee nennen nenn 12 1 9 Re packing to prevent damage during transport zureranrernn nenne nenne nennen ann 12 1 10 Description of operating elements ssssserseneenenn en en en none nn nun nun nun nun n nennen nennen 13 2 WARRANTY a eera N rennen ee arena Reese aes 16 3 START UP AND SELF TEST WITH ERROR MESSAGE AND SYSTEM SETUP 17 3 1 Printout of a self test protocol oo ccc nenn nn en nenn nn nun nun nun nun nun n nennen nennen nenne 17 3 2 Structure of the main MENU oo Ennis 18 3 3 Maintmenu display sn nn cas adie eds evel es iia 18 3 4 System setup and input of a password susessenennennnnnnnnnnnnnnnnnnnen nennen nennen nennen 18 4 QUICK MEASUREMENTS WITHOUT TEST PROGRAMMING AND STORAGE MONOCHROMATIC BICHROMATIC AND MULTICHROMATIC MEASUREMENTS KINETIC AND SCAN MEASUREMENT z2sssssssnannnnnnnennnnnnnnnnnnnnnennnnnn nenn 20 4 1 F1 NORMAL monochromatic bichro
8. 3 1 Printout of a self test protocol operating manual Upon completion of the self test after approx 5 sec the main menu appears If a printout of the results is desired press one of the following function keys within this waiting period Fi Report Evaluated test printout F2 Raw Data Printout of all raw data important for service technician Continue to main menu with F5 see diagram 17 anthos ht1 ht2 ht3 3 2 Structure of the main menu QUICK RUN TEST PLATES SETUP Fl F2 FA F5 Quick measurement Measurement of Plate storage Programming of tests and without programmed programmed tests file recall amp clear system data service test or storage L Norm Fl Choose View Fl Tests Fl Multi F2 plate Report F2 System F3 and Transm F3 Service FS test Re eval F4 Clear F5 3 3 Main menu display e The position of the plate and filter case is displayed Modifications are entered with the cursor keys e Only for HT3 Temperature control is switched on off with 0 1 on numerical keyboard e Cartridge Since no external memory expansion is installed no adjustment possible e Submenus available using the function keys F1 QUICK F2 RUN TEST F4 PLATES F5 SETUP 4 home t incub gt home eject Cartridse CEMTER k es3 measurement without test measurement with test Plate storage recall clear z setur of tests or system 3 4 System setup and input of a password e Inthe main menu press F
9. A the time the measurement was started T and the plate identification number optional The measurement values are displayed as transmission values multiplied by factor 100 and formatted right justified to 6 places preceded by at least one space Step size per measurement point 0 1753 mm STX W 450 A5 T 15 10 10 CR Start of text header 1029 2981 12902 91002 CR Result A1 H1 Position 2 18935 199 asirin iridir CR Result A1 H1 Position 1 ee CR etc 18393 9848 cccccceeeeeeeeeeees CR Result A1 Hi Position 2 17653 18384 65372 CR Result A2 H2 Position 2 ee CR etc 12903 28734 8273 897 CR Result A12 H12 Position 2 Cxx ETX 10 4 7 Checksum end of text K kinetic measurement with interval shaking Command string to the Reader example STX Khd 450 620 30 10 H 2 3 578 ETX start of text key K extended measurement range h optional bichromatic d optional time interval between kinetic cycles in sec number of measurements interval shaking H shaking mode 1 low 2 medium 3 high shaking duration maximum 60 sec optional plate identification number end of text The answer string to the computer contains at least two headers indicating the wavelength s W time the measurement was started T beginning with 0 and the number of measurement cycles N For general formatting see N normal measurement Example STX W 450 620 T OON 1 CR Start of text Header
10. NO to YES Mode of interpretation Set the type of threshold value subdivision 1 Normal Up to 4 threshold cutoffs can be programmed individually and up to 5 ranges selected 2 Auto ranging An upper and lower threshold are programmed in the next display menu The intermediate range can be divided into 2 to 10 equidistant groups 3 ranging An upper and a lower tolerance value are programmed in the next display menu The sample measurement values are printed also as percentages on the printout the sample measurement values are scaled in percent Number of ranges Only appears for mode of interpretation normal Enter the number of ranges to be subdivided minimum 2 maximum 5 e Equidistant groups Only appears for mode of interpretation auto ranging Enter the number of equidistant tolerance value subdivisions minimum 2 maximum 10 Normal Auto Ranging Ranging OD OD OD Range 2 Range 2 CV2 100 8 23 5 aD 8 5 Tr cs Qz 23 a 9 BS 3 ag z Range 1 Range 1 Group statistics No Yes The group division of measurement values and their correlations can be displayed statistically on the printout Change to YES by pressing CE operating manual 38 anthos ht1 ht2 ht3 e Data to interpret Only appears if quantitative and qualitative evaluations are combined The measured data can also be displayed in concentrations and evaluated qualitatively as such by using the standard cu
11. e Delta OD maximum slope Difference in mOD between the first measurement and the center point of the maximum slope Calculation of the center point of the maximum slope Center point between the four measurement points of the regression line with the maximum slope elta OD maximum slope OD A Delta t maximum Sope OD A Delta OD Maximum Slope centerpoint of maximum slope center point of maximum slope Delta OD Delta Time t t e Delta t relative Time in sec from the first measurement to reaching a set increase decrease in mOD e Delta t absolute Elapsed time from one preselected threshold value to an other ODA Delta t Absolute Delta Time Online Kinetic Graphic Parameters The kinetic reaction can be observed directly on the screen and is graphically displayed after measuring To optimize the display range of the curve the following parameters must be programmed Inaccurate inputs have no effect on the quality of the measurement data e OD range graphics Select a value in mOD by which the measurement data is expected to increase or decrease press CE and enter numbers on keyboard e Expected reaction Reaction process to be expected increasing decreasing e Col 1 12 Row A H Graphic display of the reaction process on the screen for a certain row row or column col during measurement Press CE select a column between 1 and 12 or a row between A and H wi
12. on the screen ates select test cursor storage enter A a E A A Number of the plate in the memory consecutive from 1 to 10 Plate ID Plate identification number combination of letters also possible It is entered prior to the measurement in the menu Run Test The incremental number of the last measured test is preset Test Test name Identification name of the measured test with the corresponding test number is programmed in the menu Test Definition Stat Status display There are three types of plate status 1 measured meas The plate was measured but the measured data have not been printed yet These plates have the highest priority in the memory mode Shift and cannot be automatically deleted 2 evaluated eval The data of these plates were printed In the shift mode they are automatically deleted consecutively from the plate memory as needed 3 special kinetic s kin A special two point kinetic has been programmed the first cycle has already been measured and it is stored under this status in the plate memory Right when the second cycle has been measured the two halves are set together and the plate gets the status meas or eval Note If no plate is to be deleted automatically the plate storage mode must be switched to PROTECT in the system setup operating manual Selection of plate with cursors Function keys Fi View All available results for this plate are dis
13. store all test parameters and tests already measured No further measurements can be performed Call a service technician N NO DEFINED TEST IS AVAILABLE TEST DEFINITION IN SETUP TESTS is displayed when a plate is to be measured in the Run Test menu but the test memory in the menu Setup Tests is empty See test programming in the chapter Define Tests NO SAMPLE SET ON PLATE EXIT WILL GIVE YOU ANOTHER CHANCE This warning is displayed if no samples were set on the plate during test programming Press F2 CONTINUE to ignore the warning NOT ALL FILTERS HAVE BEEN DEFINED is displayed if a certain number of filters to be measured was set in the Quick menu Multi but not all were allocated a corresponding wavelength Set the number of filters to the correct amount NOT ENOUGH MEMORY HEAP OVERFLOW only occurs for internal software problems i e after program dumping If the message recurs after repeated attempts to initialize the device print all programmed tests and plates already measured and call a service technician to replace the software NOT ENOUGH MEMORY PLEASE CLEAR TEST S The device s test memory is equipped with a dynamic memory structure so that although at least 30 tests can be programmed a maximum capacity cannot be defined The remaining memory available is displayed in the menu Setup Test on the lower edge of the screen Memory used xxxx bytes When this error message is displayed the remaining memo
14. un samen ana na nanannn anna anna nennen nn nn anna nen aan nn 73 10 6 8 Curve Block with multiple information 4 444RHRn nn nn nn nn en nn nun nenn nenn anne nenne 73 T0629 SRAUUS sn a ee ee a 74 10 7 Example of data transmission zersennennennennennnn nennen en enn nun nun nun nun nun nnn 74 TT MAINTENANCE Se ee ern 76 11 7 Generalsin dense ne een nn 76 11 2 Recommended regular inspection rursurerannennn eect eee nenne nenne nenne nenne nennen 76 11 3 Cle nindy s rn ee Eee ne Dele 76 11 4 Changing theibatteries n e ee un peak reden bet 77 11 5 Replacing the lamp block u2su2snnennnnnonnen nenn nennen nun nun nun nun nun nun nun nun nun nun nennen 78 11 6 Replacing the fuseSrisn oi ces ciara la Een 80 11 7 Self test Interpretation ussssennnnnonnonnennen tapii a pda api edana da 80 11 7 1 Performing the self test u een ana nn ii EEE aN 81 11 7 2 Self test interpretation nn nee anne 81 12 PRINCIPLES OF PHOTOMETRIC MEASUREMENT uuusseersnennnnnnnnennn nenn nennen 83 12 1 Units of measurement uzenssnannnnnnnnnnnnennennennn nennen EEE nun nun EEE nun nun nun nun nnnnnn nenn 83 12 2 Measurement at specific wavelengths 4444444HRHan nen an nenn nenne nennen nennen 84 12 3 Measurement range calibration 4s4 s4sHer nenn nennen nennen nn nn nnn nn nnn nennen nennen 85 12 4 Plate centering u an Ri ler ee a a eae 86 12 5 Reference ch nneln u een un Bike 87 13
15. 100 Response Formula S1 S2 2 linear regression Evaluation Interpolated concentration value of the standard point 1 113 last point of regression line ends at Conc 120 thus S1 lt 120 Function keys F1 Report F2 Transmit F4 Edit Standard Switch to screen display Standard Data F5 Select Screen 7 3 2 Standard Data operating manual Display of all standard data urve Fi ethod axis LINEAR Y axis LOGARITH A epee CEEE e Concentration programmed concentration e Response formula calculation formula for standard point e Response measured OD value e Interpolation real concentration value of the standard center point e Depending on the selected curve calculation method slope offset of regression line from the zero point of the x axis y int average deviation of the standard points from the curve correlation this value is between 0 and 1 the best curve fit quality is 1 and the axis scaling are also displayed Function keys F1 Edit All replicates of the standard previously selected with the cursor keys are displayed Standards can be removed from the calculation manually Move cursor to the desired position and press CE Renewed pressing of CE would include the replicate in the calculation again F2 Fit Modifying the curve calculation method and the axis scaling F3 Curve Switching the display screen to Standard Curve F4 Recalc Recalculati
16. 100 3 15 E06 F06 G06 H06 2 059 0 100 3 16 EO7 F07 G07 H07 2 247 0 100 3 17 E08 F08 G08 H08 2 434 0 100 3 18 E09 F09 G09 H09 2 623 0 100 3 19 E10 F10 G10 H10 2 814 0 100 3 20 E11 F11 G11 H11 3 000 3 100 3 21 E12 F12 G12 H12 3 000 3 100 3 STANDARD ID POS MEAS MEAS_STAT CONC CONC_STAT GROUP GROUP_STAT S1 A02 B02 C02 D02 142 010 S2 E03 F03 G03 H03 1 508 0 100 0 CURVE STATUS 0 FIT_MODE 0 LINEAR_REGRESSION AXIS LIN LIN CORREL 1 SLOPE 01379 YK 1285 END C B 75 anthos ht1 ht2 ht3 11 Maintenance 11 1 General WARNING The device s lamp block and the optical system located underneath are very sensitive to all light changes and thus also to outside influences such as dust particles soiling etc A few simple preventive measures can almost completely eliminate this risk PRIOR TO PERFORMING THE MAINTENANCE WORK DESCRIBED IN THE FOLLOWING CHAPTERS PLEASE MAKE SURE YOU WORK ACCORDING TO THE GUIDELINES GIVEN AT THE BEGINNING OF THIS MANUAL All maintenance work requiring opening of the instrument must be performed ony with mains disconnected and must be performed by a technical qualified person There are only a few parts in the anthos HT1 2 3 which need regular maintenance 11 2 Recommended regular inspection 11 3 Cleaning operating manual The Anthos HT1 2 3 is designed in a way that during appropriate use no danger for the user is caused The correct function with regards to measurement perfo
17. CCITT and the EIA RS232 C interface standard The following is a description of the pin configuration and the signals to be transmitted TOP APPLE SERIAL DB 9 connector plug pin configuration Signal level The range from 3V to 12V corresponds to space logical 0 for data signals and ON logical 1 for control signals The range from 3V to 12V corresponds to a mark logical 1 for data signals and OFF logical 0 for control signals The serial RS232 C interface and the Sub MIN D9 socket used for the HT1 2 3 correspond to the PC AT personal computers IBM XT25pin READER IBM AT9pin READER 10 1 DCD 1 1 XD 2 2 TT RXD 2 2 TXT RXD Er aes 3 RXD TXD 3 3 RXD DSR 6 4 DIR 4 4 GND 7 5 GND GND 5 5 GND DCD 8 6 DSR 6 6 7 CTS 9 s s EX 9 61 anthos ht1 ht2 ht3 Transmission format e 1 start bit e 2 stop bit e 8 data bits e even parity The data transmission rate can be adjusted from 300 to 19200 baud normal setting 9600 baud in the submenu SETUP SYSTEM press F5 in the main menu then F3 and F5 NEXT A precondition for the serial communication with an external computer is a terminal program which is able to receive data transmitted in the ASCII format The communication can begin as soon as the computer software is installed the serial cable connected the device setup correctly configured and the Reader in the standby mode display in the main menu The Reader s display shows
18. NEXT return to main menu with STOP e Press in sequence F5 SETUP Fi TEST and F4 DIR Printout All programmed tests including identification number measurement and reference filters measurement range selected evaluation type qualitative quantitative etc and test size in bytes will be printed Depending on their complexity at least 30 tests can be stored 6 4 Clear Erasing individual tests or the entire test memory e Go to main menu with STOP e Press in sequence F5 SETUP and Fi TEST e Mark the test to be erased with the cursor and press F5 CLEAR Either erase the selected test with F1 CLEAR or erase the entire test memory with F3 CLEAR STORAGE e Return to the test directory with F5 EXIT operating manual 44 anthos ht1 ht2 ht3 7 RUN TEST Measuring a plate with a preprogrammed test Go the the main menu with STOP Press F2 RUN TEST All parameters needed to identify a test will appear on the screen Test 4 gt TESTI Plate ID Last Fosition D 12 Dutert Channel DISPLAY select item 7 modify CE A A A E Test xx Press CE mark test in display screen with cursor and press ENTER The stored test identification number will be searched for automatically Plate ID Enter a plate identification number which can consist of a combination of letters and numbers The same plate identification number may not be given twice as long as the plate is in the plate memor
19. always formatted right justified to 5 places always preceded by at least one leading space hex 20 Generally leading zeros are displayed as spaces In case of a negative result the number is preceded by a The results are transmitted to the computer in the order A1 to H1 followed by a CR signal and the transmission of the next column Example STX W 405 T 9 13 12 CR Start header of first wavelength 1290 re re 2499 CR Result A1 H1 Did dendeveis AE deeb se adeanetien ens CR RE A ee etiaiees See Ss 145 CR Result A12 H12 W 550T 9_ 13 14 CR Header of second wavelength 59 102 T 2200 CR Result Al H1 PER ER ea e aE 2170 CR Result A12 H12 W 620 T 13 1 CR Result of third wavelength operating manual 65 anthos ht1 ht2 ht3 10 4 4 10 4 5 10 4 6 TO50 22er a 35 CR Result A1 A12 TOO Oea oetan dens 930 CR Result A12 H12 Cxx ETX Checksum end of text D Dual wavelength measurement bichromatic measurement Command string to the Reader example STX Dh 450 620 ETX start of text key D optional character h for measurements in the extended measurement range measurement wavelength reference wavelength end of text The answer string to the computer contains a header in front of the measurement results to indicate the wavelengths W the time the measurement was started T and the plate identification number optional For general formatting see N normal measurement Example STX W 45
20. are caused by reflection of electromagnetic waves striking the surface of material substances Substances absorb the complementary spectrum of their visual perceptible color Hence green plants look green since they absorb red light light of a wavelength perceived as red e g round about 750nm A photometer is an opto electronical measuring device to determine the amount of transmission of a substance for light of a certain wavelength 12 1 Units of measurement operating manual Transmission is defined as the ration between e the total available light energy at the detector measured through air and e the residual luminous energy after sample transmission at a specific wavelength Transmission is expressed as a percentage value that response in a nonlinear way to layer thickness and sample concentration Thus one layer of a certain material has a 10 transmission whereas 4 layers of the same material only have 0 01 For this reason the resulting values are converted internally into a linear measurement unit the absorption The unit is expressed in OD optical density OD log 1 trans The absorption indicates the amount of light absorbed by the sample Conversion is performed according to the Bouger Lambert Beer law The logarithm of the ratio of the light intensity before I and after the sample I expresses the result in so called OD values also Extinction E E OD log I I The optical den
21. background blue or white background Entering a password Input of a protection code for setup and or tests Modifications or deleting processes are only performable after input of the preset password e Password Setup press CE enter a 4 digit numerical password VERIFY input input repetition confirm with ENTER e Password Tests see password setup If password is activated the functions CLEAR and EDIT are not applicable anymore in the TESTS MENU NOTE If a password can not be remembered please call your local anthos distributor by means of a super user code unknown passwords can be deleted operating manual Leave the menu with F5 EXIT and return to the main menu by pressing the STOP or PREV key Instrument Setup is stored when instrument is switched off e Initial Shaking if ON the plate transport starts moving with a shake in order to stabilize the liquid meniscus without respect if additional shaking is set up in the program 19 anthos ht1 ht2 ht3 4 QUICK measurements without test programming and storage MONOCHROMATIC BICHROMATIC AND MULTICHROMATIC MEASUREMENTS KINETIC AND SCAN MEASUREMENT 4 1 Fi NORMAL monochromatic bichromatic endpoint measurement Measurement Filter Reference Filter Shak ina Shaking Duration Shakina Intensity Outeut Channel select item 3 modify CE Selection of tyre of measurement ie i anh ili iM A A E A Measurement filter select meas
22. cutoff values Cutoff NR CUTOFF CUTOFF_STAT The data fields contain the number of the cutoff NR its value CUTOFF and its status CUTOFF_STAT 10 6 7 Standards record format Display of measurement identification and evaluation data for the set standards The line shown below is wrapped Sample ID POS MEAS MEAS_STAT CONC CONC_STAT GROUP GROUP_STAT RELAT RELAT_STAT The data fields can contain the sample number ID the plate position POS the measurement value MEAS the status of the measurement value MEAS _ STAT the concentration CONC the concentration status CONC_STAT the group GROUP the group status GROUP_STAT the relation value RELAT the status of the relation value RELAT_STAT 10 6 8 Curve Block with multiple information operating manual Display of curve fit mode and corresponding data Curve STATUS 0 okay Curve status STATUS 1 invalid 73 anthos ht1 ht2 ht3 10 6 9 Status FIT_MODE 0 LINEAR REGRESSION Curve fit mode FIT_MODE 1 POINT_TO_POINT FIT_MODE 2 CUBIC_SPLINE FIT_MODE 3 4_PARAMETER FIT AXIS LIN LIN Type of axis for x and y axis AXIS LIN LOG AXIS LOG LIN AXIS LOG LOG SLOPE 0 2345 Curve coefficient for linear regression YK 2 4456 A 123 45 Curve coefficient for 4_parameter fit B 0 34E12 C 1 2234E 8 D 4 333E 14 The status is an evaluation key for the respective plate position It is only displayed in connection with measurement v
23. directory zuss4ese ren cesses nenn anne nenn nn 68 10 4 11 TR Test run plate measurement with predefined test z s4u4n HR nennen nn 68 10 4 12 H Shaking ues 2 280 san na ala apie fete ann ha wanna ana 69 10 4 13 B Transmission of plate identification number via barcode uneserserennenenennen een 69 10 4 14 R Repeat repeat of the last data transmission u4s4srererennnnn nennen 69 10 4 1775 Error messages aeiee Olid ates eh ei en BE ehe een 69 10 5 Transmission of evaluation results ersensensennenennennennenn nun nun nun nun nun nun n nennen nennen 70 10 5 1 Blocks with multiple information 4ussssrernnnnnnnnn nennen nennen nn nennen nen nennen 70 10 5 2 Blocks with the same type of information record format z2z r r nnn nennen nn 70 10 6 Description of individual DIOCKS icc cee nennen en en nun nun nun nun une nun nennen nennen 71 10 6 1 Header block with multiple information z 4 4444HHn nn nn nenn nun nn nenn ann nennen 71 10 6 2 Message block with multiple information ccccceceeeee eee e eee eee nn nn nn nennen nennen 72 10 6 3 Sample record format erri cinsi 254 Ras ance seed coon san eb ea ana nahen seated Hana nn ang 72 10 6 4 Blanks record format u ee ige De tina a panda udn 72 10 6 5 Control record format u a ea a Hude 73 10 6 6 Cutoff record format ea an aa anne ara nu 73 10 6 7 Standards record format
24. format operating manual The key words Sample Blanks Control Cutoff and Standard are used at the beginning of this type of block Depending on the number of samples these blocks always contain the same sequence of information A row of field names 70 anthos ht1 ht2 ht3 10 6 10 6 1 defining the contents of the individual record follows after the key word A record is always one line in length and contains data fields Data fields contain the information defined by their field names relating to one sample at a time Example block begin character key word for the block beginning Field names SAMPLE_ID POSITION MEAS MEAS_STAT CONC CONC_STAT Schematic transmission format blockbegin keyword 1 datadescription 1 data 1 1 datadescription 2 data 2 1 datadescription 3 data 3 1 data 3 2 etc blockbegin keyword 2 datadescription 1 data 1 1 datadescription 2 data 2 1 data 2 2 datadescription 3 data 3 1 etc blockbegin keyword 3 fieldname1 fieldname 2 fieldname 3 datafield 1 1 datafield 1 2 datafield 1 3 datafield 2 1 datafield 2 2 datafield 2 3 datafield 3 1 datafield 3 2 datafield 3 3 datafield 4 1 datafield 4 2 datafield 4 3 datafield 5 1 datafield 5 2 datafield 5 3 blockbegin keyword n fieldname 1 fieldname n etc end keyword Description of individual blocks Header block with multiple information Example Header FORMAT 110 Version number of data format PLATE_ID 123457 Plate i
25. the mains socket see drawing with a screwdriver e Remove fuse e Insert spare fuse included in supply 2 ampere slow e Close fuse carrier e Check set mains voltage next to fuse carrier turn device on and check functioning In case of malfunction call a service technician IN open cover of mains plug pull out fuse holder Y ic carefully insert 2A fuse in fuse holder lt replace fuse 250 V 2 A slow Interpretation A diode printout should be printed each time an error occurs for self interpretation and to facilitate error detection by the service technician A example for this printout is shown on the end of this chapter The HT1 2 3 by ANTHOS LABTEC has a light source consisting of 8 individual measurement channels and one reference channel which regulates the internal light control Nine individually selected tungsten halogen lamps are located on the lamp block for this purpose After the light beam has penetrated all lenses microwells and filters the residual light final transmission is focused on 9 photodetectors Their analog signal is transformed into digital values on the ADC Analog Digital Converter 80 anthos ht1 ht2 ht3 The ADC can resolve up to 2 values resulting in a theoretical range between O and 4095 We call these values Diode values because they are measured by the light detecting photodiodes on the ADC Among other things the test measurement provides the diode va
26. the provisions for the nominal range of use e the equipment has been modified or parts exchanged by a person other than ANTHOS LABTEC personnel or an authorized servicing agent e the equipment s serial number is no longer legible has been removed or altered e the equipment has not been installed in accordance with the instructions supplied by ANTHOS LABTEC e the equipment has been damaged during return transport due to false packing e g not in original packing material Only those persons may operate the equipment who are technically qualified and can interpret the data properly If a warranty is brought into operation ANTHOS LABTEC shall repair or replace any defects which have resulted from faulty material or during production as it deems fit No costs shall arise for the client All components found in the original equipment shall be available for a period of 5 five years after production This warranty contains all the obligations which ANTHOS LABTEC must fulfil and can only be amended upon the written consent of ANTHOS LABTEC operating manual 16 anthos ht1 ht2 ht3 3 Start up and Self test with error message and system setup e Place the switched off device on a suitable working surface e Connect the Epson ESC P or IBM compatible 9 or 24 matrix printer to the Centronics plug and or computer to the RS232 interface e Check the mains voltage and plug on the back of the device e Remove filter case fr
27. to an external computer e F3 LOC Moving the location cursor in the graphic display to the left F4 LOC Move the location cursor in the graphic display to the right e F5 Print Graphics Printout of all 96 grid profiles of a certain plate in the selected unit absorption transmission 23 anthos ht1 ht2 ht3 5 Test Definition Programming of tests evaluation modes operating manual DIRECTORY F1 Define Test Test Identification Pre Measurement Functions Factor Dilution Kinetic Measurement Kinetic Data Reduction Plate Layout Template Setup Report amp Quantitative Evaluation ae OO 1 Standard Points I Quantitative Evaluation Qualitative Evaluation 1 Cutoff amp Ranges L Control Test Validation y Control Validation Cee Since all programming options together are seldom used during one test optional settings may be skipped If a special point e g quantitative evaluation is required the question regarding the use of such function must first be answered with YES press CE before the corresponding menu e g standard points and quantitative evaluation is displayed This chapter will describe all of the display menus The flow diagrams will serve as an easy reference to select the program sections required 24 anthos ht1 ht2 ht3 5 1 Programming a new application test Memory free 73360 bytes us
28. two ways e Monitoring mode relative Input of a certain OD value in MOD OD increase OD decrease At the point of time when at least one of the selected wells has reached the defined decrease or increase measurement is started e Monitoring mode absolute Input of an upper and a lower threshold in MOD Again measurement is only started when at least one of the selected wells has reached this threshold Note Upper threshold value must be higher than the lower one e Read interval Select time interval for observation measurements in seconds Turn page with F5 NEXT operating manual 26 anthos ht1 ht2 ht3 5 4 Factor dilution amp kinetic measurement plate dilution factor and basic setting for kinetic measurements Kinetic Mo Interval Time Humber of Cycles ETHETIC READINGS e IHTERVAL SHAK IMG Shaking Duration Shaking Intensity select item cursor modify CE A A A e Factor dilution Press CE and enter multiplication factor to be used automatically for all measurement values OD values or data reduction values of the plate including blanks controls and standards Use the numerical keyboard The original measurement results will not be displayed any longer e Kinetic readings Change to YES with CE e Kinetic mode normal special In the normal mode up to 40 consecutive kinetic cycles may be performed automatically The special mode change with CE is only suited for two p
29. wavelengths min 2 max 8 is entered using the numerical keyboard e Wavelength 1 wavelength 2 etc press CE to display menu select filter with the cursor and press ENTER e Output channel choose from three output channels Display Measurements are displayed on the screen subsequent printout or transmission of measurement data also possible after measurement e Printer Immediate printout highest measurement of each well will be marked wavelength evaluation e RS232 Transmission of measurements to external computer Starting the measurement with F5 START MEASUREMENT 4 3 F3 KINETIC kinetic measurement k E Measurement Meas Filt Outrut DISPLAY Refr Filt nm kanae Ob Ranse ZAG mo L Interyal Reaction INCREASING Cucles Shaking VE Duration 1 Intensitas LOW select item modify CE e Measurement filter to select filter press CE move cursor to desired wavelength and press ENTER e Reference filter to select filter press CE move cursor to desired wavelength and press ENTER Usually no reference filter is used for kinetic data since a reference reduction only shifts the relevant reaction process up or down which has no effect on the results of the data reduction e Measurement range basic setting Normal Mode in Extended Mode indication limit is extended to 3 5 OD Note Wavelengths under 400 nm are not permitted using Extended Mode Measurement time is d
30. 0 620 T 12 30 01 CR Start header 1230 ne ne 2610 CR Result A1 H1 9998 werner 2003 CR Result A12 H12 Cxx ETX Checksum end of text M scan measurement of 35 measurement points per well Command string to the Reader example STX M 450 ETX start of text key M measurement wavelength end of text The answer string to the computer contains a header indicating the wavelength W and the time the measurement was started T Subsequently the measurement results are transmitted 35 transmission values per well These values are multiplied by factor 100 and formatted right justified to 6 places always preceded by at least one leading space Step size per measurement point 0 1753 mm For general formatting see N normal measurement Example STX W 450 T 14 30 20 CR Start of text header 50430 52790 52900 CR Result Al 35 measurement points Ta da NEE RR CR A900 ee 34500 CR Result H12 35 measurement points Cxx ETX Checksum end of text A Agglutination measurement with adjustable number of well positions 1 to 31 Command string to the Reader example STXA4505 2 1012ETX start of text key A measurement wavelength number of operating manual 66 anthos ht1 ht2 ht3 Example measurement points position of measurement points 15 to 15 in relation to the center of the well The answer string to the computer contains a header indicating the wavelength W the number of measurement points per well
31. 3210123456 75901234567 ttttttt set f clear CE select item modify CE e Measurement filter press CE move cursor to desired wavelength and press ENTER e Result units Measurements in transmission or absorption units change by pressing CE e Output channel Choose from three output channels Display Measurements displayed on the screen a subsequent printout or transmission of measurement data also possible after measurement e Printer Immediate printout of all values of the predefined well positions RS232 Transmission of measurements to external computer e Select locations 35 points are scanned in the well s profile 17 from the centre of the well These 35 points may be individually selected for printouts of the measurement Press CE individual move cursor down one row press CE and move point by point to the desired locations with the right cursor and select with the upper cursor Start measurement with F5 START MEASUREMENT Output channel display After each measurement the absorption or transmission profile of the well is displayed on all 35 grid points also indicating the absorption value in OD or the transmission value in for the locations selected Basic setting centre point 0 Other wells can be chosen by using the cursors The function keys provide F1 Print Printout of measurements for all selected locations F2 Transmit Transmission of measurements
32. 5 NEXT 5 8 Control amp test validation Program to evaluate the control standard and blank replicates and to automatically recognize and eliminate outliers Press CE if an automatic replicate elimination and control test validation is desired Turn page with F5 NEXT operating manual 39 anthos ht1 ht2 ht3 5 8 1 Replicate elimination Select the conditions for the automatic elimination of individual replicates operating manual This function serves to eliminate individual replicates which do not fulfil a certain programmed condition After the first elimination the mean value of the remaining replicates is recalculated and the condition re examined If necessary the elimination cycle is repeated If aminimum number of replicates to be determined is still available in the end the test is valid if not it is marked invalid on the printout If the mean value of the replicates remaining after an elimination constitutes are the basis for the test evaluation the necessary conditions are to be defined according to the next chapter 5 8 2 Control validation Condition to validate the mean value of standards controls and blanks 1 1 1 eCT Control H min number to remain A A A Program entries Column CT Definition of the standard control or blank which is to be evaluated with the following condition Press CE select the corresponding position in the display menu with the function key
33. 5 SETUP and then F3 SYSTEM 6 MAY 1996 Time S258 32 Time Mode 24h Key Click OFF Plate Storage SHIFT Internal Barcode HO select item cursors modify CE e Press F2 SLIDER EJECT remove filter case from the left side operating manual 18 anthos ht1 ht2 ht3 Check setting of e Filter position and wavelength To change delete wrong entry with CE and enter new value using the numerical keyboard Put filter case into reader e Press F1 FILTER INSERT automatic insertion of filter case e Date and time important for printed protocols Time mode 12 or 24 hour cycle e Key click to turn key clicking on or off e Plate storage shift mode a plate file already printed will be deleted automatically when the plate memory overflows 10 plates Protect mode plate files can only be deleted manually e Internal barcode NO not installed YES hardware installed To turn the page press F5 NEXT Setur Syste Printer Tare Baud Rate CRS2329 Parity RS2323 DisFlay Backaround Password Setur Passyord Tests Initial Shaking select item cursor modify CE A A E A A e Printer type choose between EPSON and IBM 9 or 24 matrix compatible printer Many ink jet printers are IBM 24 compatible except HP Deskjet which cannot be used e Baudrate proceed to selection display by pressing CE select value from 300 to 19200 bauds e Parity RS232 even or odd parity e Display
34. A E Units of concentration Definition of concentration units e g mg l important for the documentation on the printout Press CE The alphabet and a number of special characters appear in 6 rows and 5 columns in the display menu Move cursor to the row with the desired character and confirm with the corresponding function key e Curve fit method The following mathematical functions may be used to calculate the standard curve 1 Linear regression Construction of a straight line according to the least squares method with the highest possible approximation to all standard points Formula y k x d Minimum number of defined standard points 2 Linear Regression OD 0 5 0 1 0 0001 0 001 0 01 conc operating manual 35 anthos ht1 ht2 ht3 2 Point to point Direct connection of all standard points Minimum number of defined standard points 2 Point to Point OD 0 01 0 1 0 0001 0 001 conc 3 Cubic spline All standard points are connected by the best fitting curve Can only be used for nonlinear and non sigmoid functions Minimum number of defined standard points 3 Cubic Spline OD 0 0001 0 001 0 01 0 1 conc Definition of the natural cubic spline as found in G Engeln M llges F Reutter Formel Sammlung f r numerische Mathematik mit Pascal Programmen Bibliographical Institute Mannheim Vienna Zurich Wissenschaftsverlag 1985 p 146 ff 4 Parameter fit This procedure can only be used t
35. GHER THAN LOWER LIMIT appears when the wrong limit was set for either the monitoring mode or for the kinetic data reduction Delta t absolute during test programming The correct OD range must always be defined by a higher upper limit and a lower limit p PLATE DATA IS CORRUPTED CANNOT BE LOADED The plate data cannot be loaded due to a checksum error in the device s memory module program dumping If the error recurs after repeated attempts to initialize the reader please measure new plate Turn off device after 1 hour check if the complete data are still available If not replace batteries after incomplete plate memory was printed see chapter Maintenance and erase entire memory If the error occurs again call a service technician PLATE HOME POSITION NOT REACHED appears when the plate carriage does not reach its initial position on the right side of the transport system Check if transport is hindered for mechanical reasons sticky or bent guide bars Otherwise call service technician to check the home position sensor of the plate transport system and the positioning of the reflex strip PLATE STORAGE IS FULL The plate memory can store up to 10 plates including measurement data If the SHIFT type of memory was selected under Setup the evaluated plates will be erased from the memory automatically If not type of memory PROTECT the plate memory must be erased in part or completely from time to time after th
36. INTENDED USE en neh 88 13 1 Quality CONtrO lanes a e a EI BREI RR ZU 88 13 2 Galilbration na A I INES Te 88 13 2 1 Instrument Calibration veces sisia siss siaaa ak na innen han aeener 88 13 2 2 Reagent Calibration Blanking cece cece teeter ee ee nenn namen en een nn nn 88 13 3 Interpretation of measurement FESUIES cece cece cece eect estes nun nun nennen nennen 89 operating manual 4 anthos ht1 ht2 ht3 Introduction Picture 1 1 anthos ht2 The ht by ANTHOS LABTEC is a fully automated microprocessor based microplate absorption photometer Its range of application covers the universal measurement and evaluation of colorimetric endpoint specifications and kinetic tests The ht 1 covers the visible range starting at 400nm wavelength range 400nm 750nm The ht 2 covers also the UV range starting at 340nm wavelength range 340nm 750nm The ht 3 can additionally temperature control the plate to 30 C or 37 C to the accuracy required in the specification Note The temperature controlled photometers can hold micro plates at a certain temperature which can only be attained by pre incubation they are not incubators and may not be used as such operating manual anthos ht1 ht2 ht3 Warnings and Hazards operating manual General If Inflammable toxic or Biologically Hazardous substances are used when operating the equipment please observe the instructions and precautions enclosed with
37. NG5 Schott BK7 operating manual 9 anthos ht1 ht2 ht3 1 4 1 4 1 1 4 2 1 4 3 1 4 4 340 399nm lt 0 7 and 0 005 OD from 0 1 OD to 2 OD ht2 ht3 400 750nm lt 0 5 and 0 005 OD from 0 1 OD to 1 4 OD lt 0 75 from 1 3 O D to 3 0 O D e Local reproducibility at 1 O D measuring device Schott NG3 1mm 510nm lt 1 0 e Short time reproducibility at 1 O D 340 399nm lt 0 5 measuring device Hoya LA100 ht2 ht3 400 750nm lt 0 3 measuring device Schott NG3 1mm e Temperature instability for ht3 After the device is pre heated for 3 hours at 22 1 room temperature the temperature in the incubation chamber is 0 1 stable at 30 and 0 2 stable at 37 upon reaching the control point 30 minutes per temperature setting The tolerances of the measuring devices are based on the manufacturer s information General technical data Weight and Dimensions e Weight hti ht2 13 5 kg ht3 18 kg net e Charging space width x length 1700 cm bench model e Dimensions width x height x length 34 cm x 17 cm x 44 cm ht1 ht2 34 cm x 23 cm x 44 cm ht3 Environmental influences e Radio interference according to DIN VDE 0871 threshold class B for interference voltage magnetic and electric interference field force e Acoustic power level according to DIN 45 635 part 19 40 dbA e Outer lighting influences Precaution avoid direct sunlight Power supply e Consump
38. Operating Manual anthos ht I ht 2 ht 3 software version 1 23E manual version 05 02 date 06 05 2002 anthos labtec instruments Ges m b H Lagerhausstra e 507 A 5071 Wals Salzburg AUSTRIA Tel 43 662 857220 Fax 43 662 857223 email anthos anthos labtec com homepage www anthos labtec com antnes anthos ht1 ht2 ht3 Table of Contents TPABEE OF CONTEN TS iaa en ee ee een re ER Tue 2 INTRODUCTION 2a RE nme aa a Ea 5 WARNINGS gt ANB HAZARDS 2 8 Hi LE IN RL 6 1 SPECIFICATION ACCORDING TO DIN 58960 PART 4 zsssnanneennnnnnnnnnnnnnennnenn 7 1 1 General information regarding the absorption photometer ceeeeeee nenn 7 1 2 Declaration Of Conformity ccc niin aaia 7 1 3 Metrological specificati Narren An Ra e nenne nee 8 1 3 1 MEASUNING SYSTOM zu 000 anna a a a ne en an aan 8 1 3 2 Measuring processisse a na EA 8 1 3 3 DETINIELONS 32 raus ae a ee nn a Re Ra mr Tre Te 8 1 3 4 Measuring range specified according to DIN 58960 uuennesensnennnnnnnnnennnn nun nenn nennen 9 1 3 5 Rated operating Conditions 4 2 22 nsanra nenn nn nn an nn enden 9 1 3 6 Limits of error and tolerance limits eee en nn nennen nenn nenn nenn nenn nenne nennen nen 9 1 4 General technical data 2 RD ceca 10 1 4 1 Weight and Dimensions u s ususar susanne nn aan aaa ann ana nn anna 10 1 4 2 Environmental influences u esersseennneonnnnnnnnennnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnunnnnnnnnunnnnnn nenn 10
39. Setting printout format relational measurement calculation and setting the basic data for quantitative evaluation sssssssssssssrsssrrrrrrrsssr 32 5 6 1 Standard points definition of the concentration and OD values of the individual standard POINTS aier nn a er ee sete wed anal eher 33 5 6 2 How to store a standard curve eenereneeneenennnnnnnnnnnnnnnnnnnnnnnsnnnnnnnnnnnnnnnnnnnnnnnnnnnnn nenn 34 5 6 3 Quantitative evaluation Settings for quantitative evaluations such as curve calculations coordinate scaling extrapolation etC eunesenseeenennenennnn nennen une nun nenn 35 5 7 Qualitative interpretation Set to classify samples according to programmable threshold values ansehe erh eerie hes 38 5 7 1 Cutoffs and ranges Setting the tolerance values and naming the ranges 39 5 8 Control amp test validation Program to evaluate the control standard and blank replicates and to automatically recognize and eliminate outliers 39 5 8 1 Replicate elimination Select the conditions for the automatic elimination of individual replicates u chee spay digs san Bor ni nen devours rende Deere 40 5 8 2 Control validation Condition to validate the mean value of standards controls and blankssan ee na a en nenne eg 41 5 9 Safeguarding the test in the test Memory nerneanenneneen nenne een none nn nun nun nun teat nennen 42 6 EDITING PRINTING AND CLEARING OF DEFINED TESTS 43 6 1 Edit Test Changing a test already programmed
40. Y DAT SIRING START OF TEXT N A Together with the setting of the parity bit device setup this type of error control is so efficient that only a 4 fold symmetrical bit error would cause an undiscovered transmission error 64 anthos ht1 ht2 ht3 10 4 Description of commands 10 4 1 V transmission of software version number Command string to the Reader STX V ETX Answer string to the computer example STX Anthos Labtec HT2 Version 1 22 E Cxx ETX 10 4 2 F transmission of filters installed in setup Command string to the Reader STX FETX Answer string to the computer example STX 620 540 492 450 405 340 Cxx ETX Up to eight filter values can be transmitted 10 4 3 N normal measurement with up to 8 wavelengths Command string to the Reader STX Nh 3 405 550 620 ETX start of text key N optional character h for measurements in the extended measurement range number of measurement filters filter wavelengths end of text Note remember to space between individual parameters The answer string to the computer contains one or more so called headers in front of the measurement results to indicate the wavelength W the time the measurement was started T the plate identification number optional and a CR signal at the end of a line carriage return The measurement results are multiplied by factor 1000 i e transmitted in mOD Overflow values are displayed as 9999 A measurement value is
41. able and the computer s ready to receive state DO NOT USE BLANKS IN THIS FORMULA Blanks can only be used in formula calculations if their range of validity has been defined for the entire plate in the screen Plate Layout Blank Validity Plate E EXTENDED RANGE IS NOT ALLOWED FOR 340 400 NM FILTER is displayed if a filter under 400 nm is used in the extended range Either change to normal range or program another filter EXIT WILL QUIT PROGRAMMING WITHOUT SAVING THE TEST is displayed if PREV is pressed in an attempt to enter the test directory during a test definition or test edit The test editor will remain active by pressing F2 CONTINUE the test definition will be discontinued by pressing F4 EXIT modifications or any other inputs made will not be stored F FILTER HOME POSITION NOT REACHED is displayed if the filter case does not reach its home position for mechanical reasons Possible source the transport system is caught the optical sensor which recognizes the case is disaligned or defective In any case call a service technician FILTER WITH SELECTED WAVELENGTH IS NOT INSTALLED The selected wavelength is no longer installed in the device s setup Also check the correct positioning in the filter case when reprogramming I 54 anthos ht1 ht2 ht3 operating manual INTERNAL BARCODE READER Communication error between the internal barcode reader and the reader Check data format and other
42. alues MEAS_STAT concentrations CONC_STAT groups GROUP_STAT and relational values RELAT_STAT and can have one of the following values 0 okay 1 invalid 2 rejected 3 overflow 4 underflow 5 extrapolated 6 extrapolated 7 auto rejected 10 7 Example of data transmission operating manual Via the command TR a test was run remote controlled on the reader The following parameters have been set Plate layout with 4 vertical replicates horizontal filling direction and 4 blanks on position A01 to DO1 Two standards on the positions AD2 DO2 and EO3 H03 with the concentration value 1 ref 100 As a curve fit method linear regression has been selected 74 anthos ht1 ht2 ht3 operating manual Data output HEADER FORMAT 110 PLATE_ID 1000 TESTNAME TEST DATE 2 DEC 1992 TIME 14 21 52 EVALUATION QUANTITATIVE BASIS_QUANTITATIVE MEASUREMENT_DATA SAMPLE ID POS MEAS MEAS_STAT CONC CONC_STAT 1 A03 B03 C03 D03 282 0 11 110 2 A04 B04 C04 D04 420 0 21 13 3 A05 B05 C05 D05 561 0 31 4 A06 B06 C06 D06 699 0 4 5 AO7 B07 C07 D07 839 05 0 0 1 0 1 0 6 A08 B08 C08 D08 977 0 61 7 8 34 34 53 50 7 AO9 B09 C09 D09 1 118 0 71 7 0 8 A10 B10 C10 D10 1 256 0 81 72 0 9 A11 B11 C11 D11 1 394 0 91 75 0 10 A12 B12 C12 D12 1 534 0 100 3 11 E01 F01 G01 H01 1 141 0 73 39 12 E02 F02 G02 H02 1 326 0 86 82 13 E04 F04 G04 H04 1 690 0 100 3 14 E05 F05 G05 H05 1 877 0
43. ard Plate transport system Temperature display ht3 only tz Rear view operating manual 13 anthos ht1 ht2 ht3 operating manual D D DOD 1 Parallel interface Centronix 2 Serial interface RS232 3 Type plate with serial number 4 Warning For continued protection against fire hazard replace only with the same type and rating of fuse 5 Mains connector 6 fuse holder 250V 2 A slow 2 pieces 7 Mains switch 115V 230V with integrated mains filter 8 Symbol for Notice user manual 9 Warning Let instrument cool down before opening 10 Warning Before opening disconnect power cord Opening by authorized personnel only Side view Keyboard 14 anthos ht1 ht2 ht3 operating manual A A A A e e OOOO gen HOG OOO Function keys Fi F5 selection of menu and input on monitor Numerical keyboard with decimal point CE clear entry deleting and change key Cursor key ENTER Confirmation of entries PREV previous one step back to previous function or page STOP return to main menu 15 anthos ht1 ht2 ht3 2 Warranty A warranty period of 12 months shall be granted to the original buyer of the HT2 HT3 This warranty shall lose effect if e the equipment is not used in the defined scope of application e the equipment has obviously been damaged by external influences which are not in accordance with
44. arnings in the printout ensessenseneenennnnnnnnnnnnnnn nennen nennen 59 10 COMMUNICATION BETWEEN THE HT1 2 3 AND AN EXTERNAL COMPUTER 61 10 1 Principles of the communication Process nensenseneenennennnnn nun nun nun nun nun nn n nennen nenn 62 10 2 Command Keys nn sn BREI RE ini 63 10 3 Generation and transmission of checksum urssrernnnernn nenne nenn nenne nenne nennen 64 10 4 Description of commands resia aain EEE EEE Ens 65 10 4 1 V transmission of software version number uesausssnnnenennnnnnnnnnnnnn nenn nennen nn 65 10 4 2 F transmission of filters installed in setup 2z2s sasan nennen nenn nn nennen nenne nennen 65 10 4 3 N normal measurement with up to 8 wavelengths 4s4 4 4HHHn nn nn nn nn nennen 65 10 4 4 D Dual wavelength measurement bichromatic measurement eceeeeee eee ees 66 10 4 5 M scan measurement of 35 measurement points per well eseressnnnnnnnnnennennn nn 66 10 4 6 A Agglutination measurement with adjustable number of well positions 1 to 31 66 10 4 7 K kinetic measurement with interval shaking ccceeeeee settee ee eee eee ee nn en nenn nn 67 10 4 8 PD Plate directory transmission of plate memory directory c eeeeeee eee eee 68 10 4 9 PR Plate results transmission of plate results uzs4s4HHHRnnn nn nn nn nn ee nn nenn 68 operating manual 3 anthos ht1 ht2 ht3 10 4 10 TD Test directory transmission of test
45. ays on the outer casing without unplugging e Loosen air filter screw on the front of the lamp block remove screw and lock washers remove air filter with filter lattice and insert new filter e Screw filter firmly into place e Return lamp block casing into place and fasten with screw and corresponding plastic washer FILTER CLEANING EXCHANGE AIRFILTER WITH COTTON RODS SD AND OPTICAL PAPER ce 11 4 Changing the batteries operating manual The batteries which provide the turned off device with power to store tests and plate data should be replaced every two years The following signs can indicate also weak batteries Error message Selected test contains invalid data Error message Plate data is corrupted cannot be loaded Error message Not enough memory heap overflow 77 anthos ht1 ht2 ht3 e Sudden changes in the settings under the device s SETUP wavelengths time and date setting e Difficulty in printing plate data tests and memory contents Tools required Philips no 1 or Pozidrive crosshead screwdriver 3 spare batteries 1 5V alkaline format LR6 AA AM3 If instrument data is already corrupted make a printout of all remaining tests and all stored plates and delete them afterwards Note When changing the batteries as a preventive measure it is not necessary to delete the stored data e Connect a printer and switch on both instruments e Go
46. control samples only e Control well Position of the control wells before above first or after under second the sample respectively Filling direction Filling pipetting direction horizontal vertical EINEN Bil CO a apc 2 C 4 JQ 5 7 DES a Wy nn qo _ A a AL OOO OC a 2C 17 gt Coz DOC o OCL we O lt 3 ac N JC is gt 4 Gs oo Note Groups of replicates are sorted according to their replicated direction AND the filling direction Press F4 VIEW to display the current plate allocation Press ENTER to return to the display menu Plate layout functions Turn page with F5 NEXT 5 5 1 Template setup individual plate allocation with blanks standards controls and samples A E C D E F G H BLANK STANDARG CONTROL Fosition cursor set enter C e Set blanks Move cursor to desired position and press ENTER e Set standards Press Fi SET TYPE until Standard is marked The standard number is displayed at the upper right operating manual 31 anthos ht1 ht2 ht3 screen corner up to 12 different standards may be set on the plate For higher standard numbers press F2 SET for lower standard numbers press F3 SET Move cursor to the desired position and confirm with ENTER Set controls Press F1 SET TYPE until Control is marked The control number is
47. dentification number TEST_ID HIV Test identification DATE92 JAN 09 Date TIME13 01 14 Time of measurement Evaluation quantitative qualitative Evaluation mode In addition the following information depending on the test program can be displayed in a separate line Basis_quantitative measurement_data To indicate the basis of Basis_quantitative relational_factor the quantitative evaluation Basis_qualitative measurement_data To indicate the basis of Basis_qualititative concentration the qualitative evaluation Basis_qualitative relational_factor operating manual 71 anthos ht1 ht2 ht3 10 6 2 Message block with multiple information For the transmission of error messages and warnings Message message_type message_number message_text Example Message Warning 101 Standard_Status Summary of error messages and warnings Error 1 Interpretation_not_possible Error 2 Calculation_of_curve_not_possible Error 3 Quantitative_evaluation_not_possible Error 4 Calculation_of_relational_factor_not_possible Error 5 Test_invalid_Auto_replicate_elimination Error 6 Test_invalid_Control_validation Warning 100 status_of_blank_abnormal Warning 101 control_status Warning 102 standard_status Warning 110 COEFFICIENT_OF_CORRELATION_TOO_LOW 10 6 3 Sample record format Display of measurement identification and evaluation data for individual samples The line shown below is wrapped
48. der the right front corner of the block only a few rotations with the screwdriver are sufficient Evenly raise lamp block approximately 5 cm Note Do not touch lamps Loosen the two cable connections on the end of the block Note pull off on plug only Lay lamp block aside with the lamps facing up Remove new lamp block from packing without touching lamps Join plug connections Note Do not connect plugs displaced laterally and insure correct lock latches on the plug operating manual Place lamp block evenly on guide bolts and press down uniformly Tighten two screws on the top of the lamp block do not forget washers Tighten earthing screw under the right front corner of the block firmly Fasten air filter Place lamp block cover back on device Turn device on and wait for initialization Print diode values according to manual In case of malfunction check cable connections and firm fit of earthing screws If the values do not improve call a service technician Tighten screws on lamp block cover do not forget plastic washers Place the old lamp block annotating the device s 79 anthos ht1 ht2 ht3 serial number and send to the service technician in its original packing 11 6 Replacing the fuses 11 7 Self test operating manual In case of malfunction the fuses in the mains filter next to the mains socket on the rear of the device can be checked and replaced e Open fuse carrier next to
49. displayed at the upper right screen corner up to 4 controls may be set on the plate For higher control numbers press F2 SET for lower control numbers press F3 SET Move cursor to the desired position and confirm with ENTER Set samples Press F1 SET TYPE until Sample is marked The sample number is displayed at the upper right of the screen For higher sample numbers press F2 SET for lower sample numbers press F3 SET Move cursor to the desired position and confirm with ENTER For automatic sample positioning press F4 SAMPLE FILL The vacant positions are filled with samples in the prescribed filling direction If samples have been positioned manually their numbers will no longer be used To subsequently delete individual samples press CE To delete all samples press F4 SAMPLE CLEAR Turn page with F5 NEXT 5 6 Report amp Evaluation Setting printout format relational measurement calculation and setting the basic data for quantitative evaluation operating manual Wation MATRIX RELATIVE CALCULATION YES Cmodify each well by formulas CF ormula Input ur A ais E Report Format Printout Matrix Measurement printout for each plate position in 12 x 8 format Mean value calculation of blanks standards and controls only e Table Tabular printout of mean values variation coefficients deviations of individual replicates in percent concentration values and classification groups of b
50. e avoided but if necessary it must be done by qualified service personnel who are aware of the hazards involved anthos ht1 ht2 ht3 1 Specification according to DIN 58960 part 4 1 1 General information regarding the absorption photometer e Type 8 channel single beam absorption photometer with auxiliary beam path reference and discontinuous wavelength feed filter according to DIN 58960 part 2 1988 1 2 Declaration of Conformity EU Certificate of Conformity Document Nr Month Year QW 16 01 12 08 Manufacturer anthos labtec Instruments Ges m b H Address Lagerhausstra e 507 A 5071 Wals Salzburg Product Name Absorption Photometers Anthos ht1 ht2 ht3 Above mentioned product intended for General Research Lab Use Only Above mentioned product complies with the regulations defined in the following European guidelines Number 73 23 EWG Title Low Voltage Directive EN 61010 1 1993 A2 1995 EN 1658 1996 Number 89 336 EWG Title Electromagnetic Compatibility Conforms to a Minimum of Performance Criteria B EN 61326 1 1997 A1 1998 EN 61000 3 2 1995 A1 1998 A2 1998 EN 61000 3 3 1997 Corrigendum 1997 This declaration certifies the compliance with above mentioned guidelines Safety instructions included with the supplied product information have to be adhered to Issued by anthos labtec Instruments Ges m b H Place Date Wals Salzburg 21 02 2002 operating manual 7 anthos ht1 ht2 ht3 1 3 Metr
51. e data has been printed This also applies if only those plates whose status is measured remain in the memory PLATE TRANSPORT INITIALIZATION is displayed if the connection to the device s lamp block is interrupted for 57 anthos ht1 ht2 ht3 operating manual hardware or software reasons In any case call a service technician to check cable connections temperature shut down board and lamp block for proper functioning PLATE WITH SELECTED ID NUMBER EXISTS ALREADY is displayed if a plate identification number is selected in the Run Test menu that has already been allocated to a previous test For an unequivocal allocation it is absolutely necessary to assign an individual number to each plate in the plate memory which is to be measured Press ENTER and set a new ID The display is automatically preset to the next consecutive identification number PRINTER NOT READY Turm on printer check that there is enough paper in the paper feed and check for proper functioning R REFERENCE CHANNEL TOO LOW The values of the lamp block s reference channel are too low Print the diode values as described in the chapter Maintenance Measurements in the normal measurement range can usually be performed without error Nevertheless inform a service technician S SELECTED TEST CONTAINS INVALID DATA The contents of the selected test are incomplete or invalid due to errors in the device s internal memory components checksum er
52. e indication range of the instrument The specified and 0 005 OD must be taken into consideration as un linearities of the gray scale plate itself Local Reproducibility The local reproducibility proves that the measurement result of a sample is completely independent of its location in the microplate Short time reproducibility The short time reproducibility proves that the measurement result of a sample with a stable value is completely independent of the moment the measurement takes place and therefore reproducible when measuring it several times Accuracy The accuracy of the photometric measurement result proves that also the absolute photometric measurement result of a sample is within the specifications 8 anthos ht1 ht2 ht3 The specified and 0 005 OD must be taken into consideration as inaccuracy of the reference instrument 1 3 4 Measuring range specified according to DIN 58960 e Spectral absorption factor A A Normal mode 0 2 0 O D lt 400nm ht2 ht3 0 2 5 O D gt 400nm Extended mode 0 3 0 O D 400nm The limits of error and tolerance limits listed in Section 1 2 6 may be exceeded beyond this effective evaluation measuring range and for rated operating conditions other than the following section 1 3 5 Rated operating conditions e Measured wavelength 1 400 lt A gt 750 ht1 340 lt gt 750 ht2 ht3 e Blank value is deducted depending on the application under
53. ed 862 butes select test cursor storage enter A A A E E Press F5 SETUP in the main menu then press F1 TEST to continue to the directory of all tests previously programmed If a password has been programmed in the system setup enter password and confirm with ENTER or skip by pressing ENTER Press F1 DEFINE to begin programming a new test 5 2 Test identification naming filter selection and range selection operating manual select item cursors modify CE e Test ID Select an individual test identification number The first vacant position in the test memory is preset Change by pressing CE and enter number using the keyboard Press ENTER e Test name Select an individual test name May be skipped Begin setting by pressing CE When the alphabet appears on the screen use the cursors to select the row with the desired letter Press the function key F1 F5 to select the respective column Delete characters with CE Confirm the complete test name with ENTER e Measurement filter Select a measurement filter Go to menu with CE move cursor to desired wavelength and confirm with ENTER The preset filters are those set in the SETUP menu Check for accuracy e Reference filter Select a reference filter optional may be skipped Go to menu with CE move cursor to desired wavelength confirm with ENTER e Measurement range The measurement range is preset to normal range indication range O
54. f all wells according to the limits set in the test program Online Graphic Kinetic Parameters Function keys F1 Print Printout of all graphics for a certain plate F2 Col Row Choice of column or row of graphics to be displayed Move cursor to desired position and press ENTER F3 Limits Change of set display range OD Range Reaction increasing decreasing F4 Zoom Enlarging graphic display of marked well F5 Select Screen 7 6 3 Zoomed Graphics Enlarged graphic display of selected plate position Function keys F1 Print Printout of enlarged graphic F3 Zoom out Graphic display of the entire row or column F5 Select Screen 7 7 Relational Results operating manual If the option relational calculation has been selected in the test definition this screen is available in any case independent of the programmed evaluation methods The results have been calculated according to the programmed formula Function keys Fi Report F2 Transmit F5 Select Screen 50 anthos ht1 ht2 ht3 8 PLATES Plate memory re evaluation of plates without measuring printout or view of measurement and evaluation data After each measurement with a preprogrammed test all measurement and evaluation data all test parameters and the plate identification number are stored in the plate memory It can permanently store the data of up to 10 plates If the memory is full the respective error message will be displayed
55. f device software version installed filters barcode reader status and temperature control status If measurements are to be performed in the extended measurement range display range 0 3 5 OD the letter h high range will be inserted after of the command character Example STX Kh 450 15 20 H2 5 ETX kinetic measurement in the extended measurement range 450 mm measurement filter 15 seconds time interval 20 measurement cycles with interval shaking of medium intensity shaking duration 5 seconds 10 3 Generation and transmission of checksum operating manual The checksum is only transmitted from the Reader to the computer and not vice versa If it contains an error the last data string transmission can be repeated by using the R command The next command cannot be carried out until all data have been received Example data string 12345 1 char 31 hex 0011 0001 2 char 32 hex 0011 0010 3 char 33 hex 0011 0011 4 char 34 hex 0011 0100 5 char 35 hex 0011 0101 0011 0001 XOR logic Transmission of the checksum in the ASCII code 0100 defined supplement plus 0011 checksum first part gt 0100 0011 43 hex C char 0100 defined supplement plus 0001 checksum second part gt 0100 0001 41 hex A char Thus the resulting checksum is CA Thus the transmitted string is STX 12345 CCA ETX amea END OF TEXT CHECKSUM CHECKSUM KE
56. for ist cycle 1290 2343 Sieve nat 23 CR Results A1 H1 een CR etc nee 9999 22 ee eeeeee eee L45 CR Results A12 H12 W 450 620 T 30 N2 CR Header for 2nd cycle 1359 389 0 ies aana 36 CR Results A1 H1 re Vuk ewe dvaee viet CR etc W 450 620 T 300 N 10 CR Header for last cycle 2524 67 2 enenenennnnnnenennnnnn 832 CR Results A1 H1 AE A le A A CR etc 2983 782 cccceceeeeeeeeeaes 943 CR Results A12 H12 Cxx ETX operating manual Checksum end of text 67 anthos ht1 ht2 ht3 10 4 8 PD Plate directory transmission of plate memory directory Example Command string to the Reader example STX PD ETX start of text key PD end of text The answer string transmitted to the computer contains the plate status E evaluated M measured K special kinetics the plate identification number the test identification number according to test directory and the test name of each plate in the directory STX EA123 2 HERPES CR Start of text 1st plate E 1000001 12 HIV 2 CR 2nd plate M 2214991 1 RUBESTAT CR 3rd plate M 1000002 12 HIV 2 CR 4th plate Cxx ETX Checksum end of text 10 4 9 PR Plate results transmission of plate results Command string to the Reader example STX PR 100001 ETX start of text key PR plate identification number according to plate directory end of text The answer string to the computer transmits measurement and evaluation data Refer to the chapter Transmis
57. g system prior to transport for the following reasons e to minimize the influence of the sample meniscus that causes variations of the sample layer thickness within the cavity e to minimize its effect as a concave lens while determining the optical density The centering preciseness is independent of the type or the manufacturer of the used plate The plate is scanned in steps of 0 175 mm This corresponds to approximately 35 measurements per well The resulting OD values are calculated as an average of two measurement values of the 5 center points of each well 86 anthos ht1 ht2 ht3 12 5 Reference channel operating manual The device s lamp block consists of 9 independent tungsten halogen lamps 8 measurement channels 1 reference channel Reference measurements compensate possible influences on the lamp s energy during measurement as voltage fluctuations from the mains and the wear of the lamps The reference channel also serves to regulate the lamp voltages according to the used wavelength Any change of the reference value during course of measurement appears as a correction factor in the measurement 87 anthos ht1 ht2 ht3 13 Intended Use This device is intended for photometric measurements in microplates in all kind of laboratories including clinical laboratories 13 1 Quality control The quality control criteria of a diagnostic kit have to be described in the package insert of the kit For the performance
58. h of the substance Due to this fact interference filters of a narrow banded wavelength spectrum are employed in this instrument Each substance has a certain absorption spectrum A measurement should be performed by selecting the correct filter for the maximum absorption of the sample because in this way the best differentiability of various sample concentrations can be reached We can also state that the differing absorption amplitude of two uniformly composed samples is a measure for the concentration For this reason measurements in the flank area of the spectrum are not very accurate and therefore to be avoided In this 84 anthos ht1 ht2 ht3 connection it has to be taken into consideration that when using interference filters wavelength tolerances of 2 nm from the nominal wavelength value may also result in inaccuracies Each sample has an absorption minimum not specific to the measurement value which can be deducted from the measurement value automatically by choosing a suitable reference wavelength This kind of measurement is referred to as bichromatic TYPICAL ABSORBANCE SPECTRUM MEAS AND REF WAVELENGTH MEAS WAVELENGTH RER WAVELENGTH oO L ji L Ep tae LAS oo i i 4 1 L 1 L 1 J 0 T Tz T T 500 700 WAVELENGTH nm w Q Absorption spectrum measurement and reference wavelength 12 3 Measurement range calibration operating manual The device can resolve the lumi
59. he individual negative control values must be less than 0 2 OD Furthermore they may deviate from the mean value of all replicates by no more than 30 Formula X lt 0 2 AND 0 7 R1 lt X lt 1 3 R2 5 The absorption of the positive control R1 must be greater than the mean value of the first two standards S1 S2 and less than the mean value of the last two standards S3 S4 Formula S1 S2 0 5 lt X lt S3 S4 0 5 To turn page press F5 NEXT 5 8 2 Control validation Condition to validate the mean value of standards controls and blanks operating manual If a programmed condition is not fulfilled the entire test is marked invalid Press CE The following characters shall appear in the display menu e all special positions defined in the plate layout standards controls blanks e arithmetic functions Q lt gt e logical combinations NOT true if condition is not fulfilled AND true if all conditions are fulfilled OR true if one of two conditions is fulfilled XOR true if none of the entered condition is fulfilled Move cursor to select a row confirm with the corresponding function key Delete the last entry with CE Finally reconfirm the formula with ENTER z a 4 5 6 i 3 a 1 2 mmm select tyre and enter formula A A ai A E Application examples 1 The test is only valid if the absorption value of the positive control R2 i e the mea
60. he numerical keyboard and confirm with ENTER e Extrapolation The concentration of samples having a higher lower measurement results than the highest lowest standard can only be determined mathematically A continued calculation of the curve can be performed up to 40 outside the real curve fits This percentage corresponds to the concentration range between the lowest and highest standard 100 Extrapolated values are marked with an x on the printout If a curve loses its uniqueness in the extrapolation range it reaches its maximum or minimum extrapolation is blocked Press CE enter a number between 0 and 40 on the numerical keyboard and confirm with ENTER Factor conc Select a multiplication factor of up to 10 digits All concentration values of a plate can be displayed in a certain display range e Graphics report Set Yes NO to determine whether a measurement printout should contain a standard curve graph or not Turn page with F5 NEXT 37 anthos ht1 ht2 ht3 5 7 Qualitative interpretation Set to classify samples according to programmable threshold values eT ine st Muali INTERFFETATIOH Mode of interpret AUTO FAHGING Esuidistant srours 168 Grour statistics IHTERPR Data to interprete aa a vais RELATIVE VALUES select item cursor modify CE A A A A E e Interpretation If a qualitative evaluation of the samples is desired press CE The display changes from
61. ic bichromatic and multichromatic measuring with continuous scanning Types of measuring Endpoint measuring kinetic measuring and scan measuring Internal programs Memory capacity for at least 30 application protocols tests and 10 plate files identification data and test values Evaluation potential qualitative flexible threshold value input up to 4 threshold values individual group descriptions quantitative various curve fit modes point to point linear regression cubic spline 4 parameter fit kinetic data reduction computation of average slope maximum slope delta OD delta OD maximum slope delta t maximum slope delta t relative delta t absolute Relational calculations with free formula input etc Combination of all evaluation possibilities quantitative and or qualitative evaluation of concentrations values for data reduction and relation values Plates sample validation and elimination of replicates operating manual 11 anthos ht1 ht2 ht3 1 7 1 8 1 8 1 1 9 Scope of supply Warning 3 filters 620 nm 450nm 405nm with ht1 4 filters 620 nm 450nm 405 nm 340 nm with ht2 and ht3 1 filter case with lock bar 1 dust cover 1 user s manual 2 spare air filters 2 spare fuses and directions If inflammable toxic or biologically hazardous substances are used when operating the equipment please observe the instructions and precautions enclosed with such substances Never
62. is connected to the battery holder If no error can be found make a test printout and call a service technician 11 5 Replacing the lamp block Since the problems in and around the light source can have a variety of causes the lamp block should only be exchanged operating manual 78 anthos ht1 ht2 ht3 after consulting a service technician The following signs are an indication of lamp problems Error message Lamp high Maximum detection Error message Lamp low Maximum detection Error message Reference channel too low Error message during initialization test Low energy lamp energy is still sufficient to perform the measurement however a check of the lamp block is recommended OV energy lamp energy is too high check diode values of the reference channel No energy lamp energy is too low check the lamp block If the lamp block must be replaced please follow the following directions Tools required Philips no 1 or Pozidriv crosshead screwdriver spare lamp block with original packing Switch off instrument Loosen screw A on rear of lamp cover see drawing Slide lamp block cover forward and lift off Lay sideways on the outer casing without unplugging Loosen air filter screw on the front of the lamp block remove screw and lock washer lift off air filter and filter screen and lay aside Loosen two unvanished screws on the top of the lamp block and remove them Loosen earthing screw un
63. lanks standards controls and samples Combined Printout of measurement positions matrix format with separate table for blanks samples standards and controls table Relative calculation Setting all plate positions in relation to a predefined value This value is calculated according to an algebraic formula which may contain all the standards controls and blanks defined in the plate layout as well as the fundamental operations of arithmetic and numerical constants Press CE to change the relative calculation to YES 32 anthos ht1 ht2 ht3 e X Enter algebraic formula for the relative calculation X corresponds to the new value for each individual plate position Press CE The display menu shows all the standards controls and blanks defined in the plate layout as well as a variable X This variable represents all the original measurement values It must appear exactly once in each algebraic formula Move the cursor to select a row in the screen display and select the desired character with the corresponding function key Numbers may be entered on the numerical keyboard Example A negative control R1 that represents 100 binding has to be set in relation to all samples of the plate in order to deduce their amount of binding X X R1 100 Now the relational values correspond to percentage values showing the binding degree of each sample B BO calculation Quantitative evaluation Method to obtain sample co
64. lues for all 9 channels The lamp function the state of the optical system the measurement linearity etc can be checked in this manner 11 7 1 Performing the self test An automatic self test is performed during each initialization The same routine can be initiated in the service menu by pressing F1 SELF TEST The raw data is printed by pressing F2 PRINT RAW DATA 11 7 2 Self test interpretation operating manual see sample printout at the end of this chapter e ADC check Check of the proper functioning of the Analog Digital Converter board Lamp controller check Check of the proper functioning of the lamp regulation e Check of the energy for all installed wavelengths Eprom checksums Check of the software chips by checksum calculation e Diode values The diode values for all 8 measurement channels A H and the reference channel R are shown Remove the filter case from the device press the right cursor in the main menu Check the number of filters filter position and wavelengths by referring to the protocol Vacant positions in the filter case should not be programmed 0 to avoid error messages such as Division by Zero better position 5 5 position 6 6 etc The first part of the diode values indicates the so called bright values They should lie between 2500 and 4000 Vacant positions must be displayed as 4095 i e Overflow since the maximum lamp energy white light strikes the detect
65. matic endpoint measurement s es 20 4 2 F2 MULTI measurement with several wavelengths simultaneously 21 4 3 F3 KINETIC kinetic measurement ernerseneeneenennennenn none nn nun nun nennen nennen nennen 21 4 4 F4 SCAN scan and agglutination measurements eceeeeeeee eee nenn nenn 23 5 TEST DEFINITION PROGRAMMING OF TESTS EVALUATION MODES 24 5 1 Programming a new application test ccc ceeeee eee nenn nenn nennen nennen nennen 25 5 2 Test identification naming filter selection and range selection 25 5 3 Premeasurement functions optional parameters for functions being performed prior to measurement are br nr nenne 26 5 4 Factor dilution amp kinetic measurement plate dilution factor and basic setting for kinetic meas rements nn rl een 27 5 4 1 Kinetic data reduction Selection of data reduction method and parameters for screen graphics during kinetic measurement unusesesnenennnnnnnnnnnnnnnn nennen nun nennen nenn nenn 28 5 5 Plate layout functions Setting parameters for the validity of blanks filling direction replicates and control strips ursssssnennnn nun nun nun eee eset nennen nennen nennen 30 operating manual 2 anthos ht1 ht2 ht3 5 5 1 Template setup individual plate allocation with blanks standards controls and SAMPIES cur steady ces san ae des and cob kanes BE biodata code dees Estab sees a ee 31 5 6 Report amp Evaluation
66. menu Setup Template B BARCODE READ ERROR is displayed if the barcode was set to YES in the device s setup a measurement was started in the menu Run Test but the barcode label for the plate identification cannot be read Check if the plate in the plate transport has a barcode if the label has been properly attached and if the hardware for an internal barcode reader has actually been installed Only the following high density codes can be read EAN8 EAN13 CODABAR CODE 39 UPC A UPC E INTERLEAVED 2 5 CODE 128 only on special request Either restart with F2 REPEAT enter the plate identification number manually with F3 MANUAL INPUT or return to the main menu with F4 SKIP 53 anthos ht1 ht2 ht3 operating manual C CONTROL MEASUREMENTS WRONG MIN CAL MAX VALUES signifies an error during a measurement in the extended range Check the filter inputs the diode values of the lamp block and the cleanliness of the grey glass at the end of the plate transport If the error recurs call a service technician CUTOFF FORMULA MUST BE DEFINED is displayed if a certain number of cutoffs was programmed for the qualitative interpretation but no valid cutoff constants or formulas were programmed Refer to the chapter Test Definition Qualitative Evaluation D DEVICE NOT READY Upon being activated externally the computer does not send a Clear to send CTS signal Check hardware connections c
67. n value of its replicates is less than or equal to 0 8 OD Formula R2 lt 0 8 2 The test is only valid if both controls lie within the linear range of the photometer According to the specification the linearity of READER 530 as of 0 1 OD is defined as 1 at 450 mm Formula R1 gt 0 1 AND R2 gt 0 1 41 anthos ht1 ht2 ht3 The test validation is completed Entries may still be modified by turning back the pages with PREV Turn page with F5 NEXT 5 9 Safeguarding the test in the test memory To safeguard the test just programmed in the test memory press F2 The test is to be found in the test directory now If the test is not to be stored press F4 NO This display window does not permit turning back the pages with PREV A new test has_been defined es Test name TEST bo you want to store this test Cyes noo operating manual 42 anthos ht1 ht2 ht3 6 Editing printing and clearing of defined tests 6 1 Edit Test Changing a test already programmed With the reader turned off Turn on the reader After completion of the self test press F5 EXIT in the main menu press F5 SETUP and then F1 TEST The screen will display a directory of all the programmed tests Mark the test to be edited with the cursor Confirm by pressing F2 EDIT With the reader turned on Press STOP The main menu will appear on the screen Press F5 SETUP and then F1 TEST to display all tests already pr
68. ncentrations using a standard curve Number of standards Depending on the type of curve calculation a minimum number of standards must be set Linear regression 2 standards Point to point 2 standards Cubic spline 3 standards 4 parameter fit 4 standards Maximum number of standards 12 Enter number on numerical keyboard and press ENTER A quantitative evaluation can be performed independent of the standards defined in the layout e Basis for quantification Appears only if relative calculation YES was selected Set according to whether the original measurement values or the results of the relative calculation relative values are to be used for the quantitative evaluation Turn page with F5 NEXT 5 6 1 Standard points definition of the concentration and OD values of the individual standard points operating manual e Standard number Only the number of standards set under Report and Evaluation may be used for evaluation The number of standards defined in the plate layout is not relevant for the actual number of standard points of a curve 33 anthos ht1 ht2 ht3 Concentration value x Enter the concentration value to be plotted on the x axis of the curve graphic Constants of up to 12 digits may be entered on the numerical keyboard Response value y Enter the absorption value to be plotted on the y axis of the curve graphic It usually corresponds to the OD values of the standards on the plate
69. nded range are too low Measurements in the normal range are possible Check the calibration glass on the left rear edge of the plate case for dirt collection and streaks Clean as well as possible Call a service technician LAMP LOW MAXIMUM DETECTION The lamp power on the detector is too low to be able to perform a precise measurement Print the diode values as described in the chapter Maintenance check the installed filter and its program in the setup if necessary the filter and the lamp block must be replaced Call a service technician Check the optical system for dirt collection liquid residues particles etc LAMP LOW MINIMUM DETECTION During a calibration measurement the minimum darkness value has not been attained This is usually due to dis adjustments on the ADC board amplifier offset detector diodes and can only be rectified by a service technician 55 anthos ht1 ht2 ht3 operating manual LAYOUT PARAMETERS MODIFIED CONTINUE WILL CLEAR TEMPLATE is displayed when the automatic plate layout functions are to be modified during a test edit This also involves a new individual plate layout Press F2 to delete the previous plate layout press F4 EXIT to discontinue the modification attempt M MEASUREMENT NOT POSSIBLE TRANSPORT SYSTEM NOT INITIALIZED occurs when the plate transport could not initialize properly If this message recurs after repeated attempts to start or self tests in the service menu
70. nous energy received by the detector into a maximum of 4095 values diode values Prior to each measurement a calibration is performed automatically by measuring the dark value for 0 transmission and the bright value for 100 transmission blank measurement against air The resulting values correspond to the resolution limits which can be obtained in practice The device has been pre adjusted such that the diode values are greater than O for the dark value measurement and lower than the theoretical maximum for the bright value measurement 4095 This prevents an underflow or overflow of the theoretically possible limits This initial calibration always guarantees a 100 measurement range even if there is a change of the luminous energy 85 anthos ht1 ht2 ht3 4095 100 MAX Bra ow O 3 uw a ie 2 s MEAS MIN MEAS 2 Z MAX MIN 2 Z lt A k 0 MIN o0 The formula to calculate a transmission value is as follows T Meas Min Max Min T Transmission Meas Diode value during measurement Min Diode value at 0 measurement Max Diode value at 100 measurement Example 1 Max 3600 Min 20 Meas 30 30 20 3600 20 0 0028 Log 1 0 0028 2 552 OD Example 2 Max 3600 Min 20 Meas 3000 3000 20 3600 20 0 754 Log 1 0 754 0 122 OD 12 4 Plate centering operating manual The microplate is centered precisely under the optical axis of the measurin
71. ntensity from three levels low medium high Maximum filling capacity per well 200 ul maximum shaking duration 99 sec Turn page with F5 NEXT 5 4 1 Kinetic data reduction Selection of data reduction method and parameters for screen graphics during kinetic measurement operating manual Data reduction Selection of data reduction method Go to display menu with CE Choose from the following parameters e Delta OD Difference between two kinetic measurements in MOD OD OD 2 Delta OD OD 1 e Average slope Slope in mOD min between the first and last kinetic cycle Only suited for linear reaction progressions with few cycles and measurement points e Maximum slope Maximum slope of the curve in mOD min Maximum reaction speed e Type of evaluation Linear regression lines connect four measurement points at a time line through measurement points 1 2 3 4 then through 2 3 4 5 through 3 4 5 6 etc The line with the highest slope is calculated The accuracy of this calculation depends on the number of measurement cycles selected Number of regression lines number of measurement cycles v 3 see diagram Maximum Slope mOD min center point of maximum slope 28 anthos ht1 ht2 ht3 operating manual e Delta t maximum slope Time difference in sec between the first measurement and the occurrence of the center point of the maximum slope
72. ntrols and standards on the plate Set the following parameters e Direction Direction horizontal vertical of replicates Sad S48 Sap SABE SIE S4BL 5 S Ss 5 ss ss 5 58 A A A A A A A A A A A SdBL Sde SIBL SABL SAAL SSB By BL BL BL BL BL BL BL BL BL BL BL S4BL S4BL SBL SdBL S4BL S lt 4BL 6S ES EEE S 8 8S 5 A A A A amp A A A A A A A SdB SdB SIBL Sd SOB S lt 4BL BL BL BL BL BL BL BL BL BL BL BL BL SJB SIR Sda SAaBL SARL SI s 55 8 5 8 8 8 A A A A amp A A A A A AA SABL 54B SAAL S4BL S lt ABL S lt IBL BL BL BL BL BL BL BL BL BL BL BL BL SABL S48 SAB SABL SdH SSB 5s 5 5 S 8 8 A A A A A A amp A A A AA SAaBL S lt 4BL S4BL S4BL SARL SSB BL BL BL BL BL BL BL BL BL BL BL BL Number of wells Number of replicates maximum of 12 replicates for horizontal direction maximum of 8 replicates for vertical direction e AG control strips Antigen control strip setting A control sample which is automatically deducted is assigned to each sample The sample and control sample have the same numbering on the printout The control well sample positioning depends on the filling direction and the sample control setting combination of the reader 30 anthos ht1 ht2 ht3 Note AG control strips cannot be used together with the automatic replicates Set the following parameters e Direction Direction horizontal vertical i e one row column consists of samples only the next row column consists of
73. nvalid since a condition for control validation was not fulfilled An attempt to evaluate is made nevertheless Find error by means of the printout WARNING BLANK STATUS At least one replicate of a blank is faulty overflow rejected invalid auto eliminated The test is evaluated A check of the printout is recommended WARNING CONTROL STATUS At least one replicate of a control is faulty overflow rejected invalid auto eliminated The test is evaluated A check of the printout is recommended WARNING STANDARD STATUS At least one replicate of a standard is faulty overflow rejected invalid auto eliminated The test is evaluated A check of the printout is recommended WARNING QUANTITATIVE EVALUATION COEFFICIENT OF CORRELATION TOO LOW The correlation coefficient has been defined as a measure for the fit of the individual standard points in the calculation mode of the linear regression lines The best curve fit is defined as 99 points Iying on the line In the test definition Quantitative Evaluation a minimum coefficient was defined which falls short of the previously run test A possible remedy is to strike out replicate outliers manually if no automatic replicate elimination or an insufficient automatic replicate elimination was programmed 60 anthos ht1 ht2 ht3 10 Communication between the HT1 2 3 and an external computer operating manual The serial interface is specified according to the
74. o characterize sigmoid curves Minimum number of defined standard points 4 The curve is calculated according to the following formula y a d 1 X c d 4 Parameter Fit OD 0 1 0 0001 0 001 0 01 conc operating manual 36 anthos ht1 ht2 ht3 operating manual Definition The dose level which results in a response exactly half way between the zero dose response a upper asymptote and the infinite dose response d lower asymptote is designated as the midrange of the assay and is designated c In addition the curve is characterized by a slope factor or exponent designated by b Definition according to Radio Immunoassay and Related Procedures in Medicine International Atomic Energy Agency Vienna 1974 Volume 1 p 165 ff The axial scaling is preset in the 4 parameter fit and cannot be adjusted e x axis scale Scaling the x axis linear logarithmic Change with CE e y axis scale Scaling the y axis linear logarithmic Change with CE e Minimum correlation Only valid for curve fit method linear regression Method to control a certain curve quality Setting a divergence coefficient between 1 and 99 of the individual measurement points from the real curve to control a certain curve quality Best fit point lying on the curve is 99 If the preset limit of individual points is exceeded the respective warning appears on the printout Press CE enter number between 1 and 99 on t
75. o be deleted and confirm with F4 SELECT If previously marked well is to be included in the computation again move the cursor to the desired position and press F2 CLEAR or F1 CLEAR ALL Confirm with ENTER 46 anthos ht1 ht2 ht3 F5 SELECT SCREEN Depending on the programmed evaluation methods different ways of displaying the results are available This function is available for all other program sections of this submenu 7 2 Plate Layout A B D E F G H The plate layout with all special positions blanks standards controls and the numbered samples set in selected test are shown on the display Function keys F1 REPORT F2 TRANSMIT F5 SELECT SCREEN 7 3 Quantitative displays For the test results of a quantitative evaluation the standard curve is always displayed first Its computation may take a few minutes above all in the case of the 4 parameter fit A counter on the display shows that the reader is still working 7 3 1 Standard Curve operating manual First the curve is computed according to the programmed Curve Fit Mode and displayed on the screen In the case of a linear regression the calculation limits for the measurement values are established by the calculated straight line Thus concentration values can be displayed as lt Minimum although they are greater than the programmed minimum concentration 47 anthos ht1 ht2 ht3 Example Standard point 1 Concentration
76. of any reagent quality control it has to be referred exclusively to those instructions With an internal Control Validation see chapter 5 8 the software provides optionally a mean to program such quality controlling processes in the HT1 2 3 13 2 Calibration 13 2 1 Instrument calibration In some kit instructions the instrument calibration is referred to as blanking against air This is equivalent to the automatic instrument calibration described in chapter 1 3 2 Measuring process 13 2 2 Reagent Calibration Blanking operating manual Generally reagent calibration is done by deducting a blank value from sample control and standard values See chapter 1 3 5 Rated operating conditions The programming of blanks has to be done according to the instructions given in the package insert of a diagnostic kit 88 anthos ht1 ht2 ht3 13 3 Interpretation of measurement results operating manual The interpretation of measurement values is subject to the limits of error and tolerances See chapter 1 3 6 Limits of error and tolerance limits Measurement data in the field of vertical photometry are dependent on liquid volumes meniscus and microplate shapes Therefore they are relative values and evaluation is possible only within one microplate For the interpretation of results it has to be referred to the instructions given in the package insert of a diagnostic kit It is the user s responsibility to check evaluated
77. ogrammed Mark the test to be edited with the cursor and confirm by pressing F2 EDIT Screen display input and delete functions of the Edit test are identical to test programming Modifications will not be executed until the storage procedure has been carried out 6 1 1 To copy a test Select the test to be copied in the test directory The next display screen will show the parameters for the test identification Select the test ID current identification number in the test memory with the cursor Press CE and enter a new unused identification number for the test copy on the keyboard confirm by pressing ENTER If the test copy should not be stored under the same name move the cursor to Test Name press CE enter the new name and confirm with ENTER Edit the copy and press F5 NEXT up to the inquiry Store changes YES NO Press F2 YES 6 2 Print Test Printout of all programmed test parameters operating manual Go to main menu with STOP Check printer settings under System Setup Press F5 SETUP and then F1 TEST Use cursor to select the test whose parameters are to be printed Turn on printer and press F3 PRINT Printout All parameters programmed for the test including plate layout printing date and time will be printed 43 anthos ht1 ht2 ht3 6 3 Dir Printout of the entire test directory e Go to main menu with STOP e Check printer inputs under System Setup F5 SETUP F3 SYSTEM F5
78. oint kinetics with long interval time The plate is stored with the status s kin in the plate memory submenu PLATES after the first kinetic cycle The reader is immediately available for other measurements A second cycle can be run after any period of time by recalling the test in the menu RUN TEST Note Performing special kinetics ensure identical plate ID number of both cycles Both measurements will automatically be evaluated as one special kinetic reading The measurement results will be in Delta OD values mOD The following parameters will only be visible if kinetic readings YES was selected e Interval time Note interval time setting in seconds from the beginning of one measuring cycle to the beginning of the next cycle maximum time 36000 Minimum Interval Time Normal Mode Single Wavelength 5 sec Single Wavelength Shaking 7 sec Shake Duration Dual Wavelength 10 sec Dual Wavelength Shaking 12 sec Shake Duration Extended Mode Single Wavelength 10 sec Single Wavelength Shaking 12 sec Shake Duration Dual Wavelength 20 sec Dual Wavelength Shaking 22 sec Shake Duration e Number of cycles number of measurement cycles to be performed max 40 Press CE enter number on keyboard confirm with ENTER operating manual 27 anthos ht1 ht2 ht3 e Shaking Shaking of plate before each measuring cycle Change to YES with CE enter duration in seconds using keyboard select shaking i
79. ological specification 1 3 1 Measuring system Pinhole Well Detector IR filter Beam path see diagram Radiator preadjusted automatically controlled Tungsten halogen lamps 1 lamp per channel spectral apparatus dielectric interference filter filter cartridge expandible for up to 8 filters standard components 620 nm 450 nm 405 nm 340 nm only ht2 and ht3 Other wavelengths available upon request Radiation receptor silicon photodiodes Store for objects to be analyzed all 8 x 12 microtitration plates with or without vertical subdivision Indication size A X spectral absorption factor Indication range 0 2 5 A lt 400 nm only ht2 ht3 0 3 0 A gt 400 nm normal mode 0 3 5 A gt 400 nm extended mode Indication type Digital display Speed per measurement monochromatic 7 sec bichromatic 10 sec kinetic cycles 5 sec each 1 3 2 Measuring process 1 3 3 Definitions operating manual Measuring class index absorption corresponds to O D optical density measuring geometry see diagram 1 Zero point setting self calibrating Concentration test comparison value process Measured value test reference value process Control measurements automatic prior to each measuring operation self test after switching on Linearity The linearity is measured with a gray scale plate with linear increasing OD values This is done to prove the linearity of the measurement results within th
80. om the additional carton and check firm support of filters and lock bar e Connect power cable to standard mains plug e Switch on main switch left bottom The Reader performs the complete initialization including self test in approx 40 seconds e When the display No plate carriage detected appears press ENTER e When the display Please insert filter slider appears push the filter case into the right side filter carriage to the end stop and press any key e This automatic self test re calibrates the device and checks if the electronic and mechanical components are in the proper service condition The following error display may appear e System halted call service technician Lamp controller check not OK call service technician Lamp controller check error check power supply call service technician ADC check not OK call service technician Plate check not OK Check microtitration plate in the plate transport otherwise call service technician e Check filter slider Check filter slider in the device for position and completeness otherwise call service technician No filter detected No filter in this position check filter adjustments OV energy low energy no energy check filter adjustments and positions otherwise call service technician For further information on error interpretation and correction press F3 after the self test has been performed By pressing F5 the reader goes to the main menu
81. on of the test If test parameters have been modified this will be performed automatically upon switching the display screen F5 Select Screen Choice of all available display screens 48 anthos ht1 ht2 ht3 7 4 Quantitative Results Display of real concentration values for all plate positions Blanks are marked Concentration values less or greater than the calculated minimum or maximum value respectively are marked lt or gt if no extrapolation value was programmed Extrapolated values are marked with an x in front of the value Instead of automatically eliminated values A REJ auto rejected is displayed Function keys Fi Report F2 Transmit F5 Select Screen 7 5 Qualitative Results ne SE en 1 2 5 6 A D E F G H Group layout of the individual plate positions Blanks are marked instead of automatically eliminated values A REJ auto rejected is displayed Function keys F1 Report F2 Transmit F5 Select Screen 7 6 Kinetic results displays 7 6 1 Kinetic Raw Data operating manual For kinetic measurements the internal absorption values for each measuring cycle are only stored up to the next kinetic or scan measurement therefore a printout should be made The cycles may be paged by using the cursor Function keys Fi Report F2 Transmit F5 Select Screen 49 anthos ht1 ht2 ht3 7 6 2 Graphics 8 12 Display of the reaction course o
82. or Thus the first position without a filter does not indicate an overflow value since normally no light can reach the detector due to the filter s lockbar Usually the diode values decrease slightly for filters with shorter wavelengths This is due to the fact that for filters with shorter wavelengths more energy is needed This has no effect on the measurement results e The second part of the diode value printout indicates the so called dark values They are attained by measuring with turned off lamps and correspond to the minimum measurement values For wavelengths greater than 400nm they must lie between 10 and 50 for wavelengths less than 400nm between 30 and 100 This difference is due to a built in amplification switch e Check ADC A further check of the proper functioning of the amplifier switch for wavelengths less than 400 nm The 81 anthos ht1 ht2 ht3 values for the first line should lie between 10 and 50 for the second line between 30 and 100 e Check lamp controller The values of the first line must clearly deviate from those of the second line at least 100 diode values Example for a selftest printout of the anthos HT Anthos HTII Reader Version 1 22 Self Test Date 30 APR 1999 Time 7 32 29 Page 1 SEr Tal Nr ar A ar Mara OM arana naar ADC check OK Lamp controller check OK 1 Filter 620nm Energy for filter OK 2 Filter 540nm Energy for filter OK 3 Filter 492nm Energy for filter OK 4 Filter 450nm Energ
83. oubled e Interval interval setting in seconds from the beginning of one measurement cycle to the beginning of the next cycle maximum time 36000 seconds operating manual 21 anthos ht1 ht2 ht3 operating manual Minimum Intervals Normal Mode Single Wavelength 5 sec Single Wavelength Shaking 7 sec Shake Duration Dual Wavelength 10 sec Dual Wavelength Shaking 12 sec Shake Duration Extended Mode Single Wavelength 10 sec Single Wavelength Shaking 12 sec Shake Duration Dual Wavelength 20 sec Dual Wavelength Shaking 22 sec Shake Duration e cycles number of measurement cycles to be performed max 40 press CE and enter number using keyboard e Shaking plate shaking during each measurement cycle press CE and change to YES enter duration in seconds using keyboard choose shaking intensity from three levels low medium high e Output channel choose from three output channels Display Measurements are displayed on the screen subsequent printout or transmission of measurement data also possible after measurement e Printer Immediate printout for the printout format select either raw data printout of measurement data for each cycle or maximum slope maximum reaction slope in mOD min e RS232 Transmission of measurements to external computer The kinetic reaction progress can be viewed directly on the screen and is displayed graphically after measurement To optimize the di
84. played on the screen The data may also be printed or transmitted to an external computer This submenu is identical to the menu appearing after completion of a measurement Run Test under Output channel Display 51 anthos ht1 ht2 ht3 F2 Report Printout of a report in the matrix table or combined format depending on the test program setting F3 Transmit Transmission of measurement values to an external computer F4 Re eval Repeated evaluation of the raw plate data without measurement This function makes it possible to have the measurement values of a plate evaluated with another test or to have two or more tests performed on one plate If a plate is to be evaluated with a new test make sure the setting of blanks controls standards and replicate numbers etc conform with the layout programmed for the new test This function is normally used to edit small programming errors in the test and to have the plate data re evaluated without having to remeasure the entire plate Move cursor to mark the plate to be re evaluated press F4 Re eval press CE to select a new test from the display screen and confirm with ENTER Set the last position to be measured on the plate under Last Position and select an output channel Press F5 START EVAL After the plate has been evaluated the data can either be displayed on the screen printed or transmitted to an external computer In the plate memory the plate will remain linked
85. results with the corresponding raw data values 89
86. rmance is assured by an automatic check of the relevant technical parameters prior to each individual measurement In case of a problem the instrument reports an error message Anthos recommends to have the instrument inspected by an authorized service partner at an interval of about 2 years During this inspection the correct function of the instrument can be verified Additionally Anthos offers special test plates which may be used by the user to regularly verify the performance of the instrument 1 Always cover the turned off device with the dust cover included in supply 2 Clean filter Turn on device press right cursor remove filter case from the right insert Press sides of locking bar together carefully slide bar and filter from the case Blow with compressed nitrogen and clean manually with a cotton 76 anthos ht1 ht2 ht3 DUSTCOVER a swab or optical paper Do not use alcohol or detergents containing water Return filter to its case and check the order in the device s setup Push filter case into the filter transport system till it reaches the end stop and press left cursor The filter case moves to its operating position Change air filter every four to six months Tools required Philips no 1 or Pozidrive crosshead screwdriver spare dust filter as included in supply e Loosen screw A on rear of lamp casing see drawing next page e Slide lamp block casing forwards and lift off Lay sidew
87. ror Nevertheless try to print the test parameters If only one test is affected by this error message do not erase otherwise the defective memory area could spread to other tests SERIAL COMMUNICATION The device is having trouble in serial communication with a computer Check if the baud rate parity bit and data format conform in both devices data is received with frame or overflow errors SERIAL COMMUNICATION BUFFER FULL The maximum length of the data string sent by the computer may not exceed 192 bytes SHAKING DURATION TOO LONG FOR DEFINED MEASUREMENT Either the shaking duration must be shortened or the interval time extended See drawing Minimum interval time in chapter Factor dilution amp kinetic measurement T TRANSPORT STEPS LOST The measurement value evaluation on the microtitration plate is defined by a certain number of steps Steps can be lost due to an inaccurate plate transport and a plate evaluation is no longer possible If the error message recurs call a service technician 58 anthos ht1 ht2 ht3 W WRONG X DEFINED CANNOT USE X The formula in the validation part of the test definition must not contain an X incomplete erasing of XOR WRONG X DEFINED MUST BE USED ONCE A formula must be entered for the evaluation of relational values in the test definition in which the variable X representing all samples controls standards and blanks on the plate occurs exactly once This i
88. rve If a relational calculation was programmed these values can also constitute the calculation basis Turn page with F5 NEXT 5 7 1 Cutoffs and ranges Setting the tolerance values and naming the ranges e Cutoff values for measurement results concentration values relational values Enter the formula or contents for the programmed tolerance values Normal up to 4 cutoffs can be programmed auto ranging or ranging two cutoffs must be programmed Press CE All standards controls and arithmetic functions which can be used in the cutoff formula appear in the display menu Enter numbers with the keyboard Move the cursor to select a row in the display menu and confirm the desired character with the corresponding function key Confirm complete formula by pressing ENTER Naming the groups defined by the preset cutoff values e Range 1 x lt CV1 All values falling in this range must be less than the first cutoff CV Press CE to display the alphabet in 6 rows and 5 columns Move the cursor to select a row and confirm the desired character with the corresponding function key Enter numbers with the keyboard Confirm again by pressing ENTER etc If mode of interpretation auto ranging or ranging the intermediate range between the two cutoff values cannot be individually named since either the group number or a percentage is automatically printed as the name This range is referred to as special Turn page with F
89. ry is not sufficient for a new test definition After printing the test parameters erase one or more tests move cursor to mark desired test press F5 CLEAR confirm with F1 ID xxx NOT ENOUGH STANDARDS DEFINED FOR THIS CURVE FIT METHOD The minimum number of standard points for various curve evaluation methods linear regression 2 points point to point 2 points cubic spline 3 points 4 parameter fit 4 points refer to the chapter 5 5 Report amp Evaluation 56 anthos ht1 ht2 ht3 operating manual NO PLATE DETECTED No microtitration plate was detected in the plate transport Insert a plate and start again If this error message occurs frequently for no apparent reason inform a service technician NO TEST WITH THE SELECTED ID NUMBER IS AVAILABLE appears when a test was selected with a test identification number for which no defined test is yet available in the Run Test menu Press ENTER move the cursor to the right on the test name and press CE A list of all the programmed tests with the corresponding identification number will appear Set the cursor to the desired test and press ENTER NO WELL POSITIONS SET FOR MONITORING During test programming at least one plate position whose value is to be compared with the defined parameters must be selected in the Monitoring Mode Go to Well positions press CE and select the desired position on the plate matrix displayed on the screen O OD UPPER LIMIT MUST BE HI
90. s confirm with ENTER e Column N Enter the minimum number of remaining replicates Use keyboard for numerical entries and confirm with ENTER Column Condition Enter one or more conditions which must be fulfilled by each replicate to avoid elimination Press CE a display menu appears on the screen with the following character options for formula selection all special positions defined in the plate layout standards controls blanks arithmetic functions lt gt e logical combinations NOT true if condition is not fulfilled AND true if all conditions are fulfilled OR true if one of two conditions is fulfilled XOR true if none of the entered condition is fulfilled e X represents each individual replicate of a certain control blank or standard Move cursor to select a row and confirm with the corresponding function key Delete the last entry with CE Finally reconfirm the formula with ENTER Application examples 1 The absorption of a blank value may not exceed 0 020 OD Formula X lt 0 02 2 The absorption of the individual negative control value R2 must be less than or equal to 0 150 OD otherwise the 40 anthos ht1 ht2 ht3 replicate must be eliminated Formula X lt 0 15 3 The deviation of the individual standard values of S1 may exceed the mean value of all standards of S1 by no more than 20 Formula S1 0 8 lt X lt S1 1 2 4 The absorption of t
91. s necessary to limit the relational calculation to a linear model Also refer to the chapter Report amp Evaluation WRONG X DEFINED YOU MUST USE X For the automatic elimination of replicates in the test definition a formula must be defined for each control each blank and each standard to be evaluated in which the variable X represents the individual replicate of the control the standard the blank must be used at least once 9 2 Error messages and warnings in the printout operating manual ERROR CALCULATION OF RELATIONAL FACTOR NOT POSSIBLE The relational factor is calculated with a formula programmed in the test definition If an element of such formula becomes invalid after completion of the replicate elimination or test validation etc e g S1 was rejected or R1 is overflow no relational factor can be calculated ERROR INTERPRETATION NOT POSSIBLE No qualitative evaluation can be performed This error message occurs if the values of the cutoffs are not in ascending order or if the cutoff formula cannot be calculated at all e g since an element in the formula is nonexistent overflow rejected etc ERROR INVALID CURVE SHAPE is displayed if the standard curve cannot be calculated unequivocally Possible remedy Select another curve fit mode ERROR NO BLANK SUBTRACTION POSSIBLE If all replicates of a certain blank are faulty overflow invalid a blank subtraction is no longer possible No evalua
92. setup inputs on the Barcode Board INTERVAL TIME TOO SHORT FOR DEFINED MEASUREMENT For the minimum time interval setting refer to the drawing in the handbook Factor Dilution amp Kinetic Measurement INCOMPLETE RESULTS CONTINUE EVALUATION This warning is displayed when a kinetic measurement was aborted before all measurement cycles could be completed The evaluation can either be aborted or continued with the incomplete data L LAMP HIGH CALIBRATION FILTER Calibration filter measurements in the extended range are too high Measurements in the normal range are possible Check the calibration glass at the rear left edge of the plate case for mechanical damage Call a service technician LAMP HIGH MAXIMUM DETECTION The lamp power on the detector is too high to be able to perform a precise measurement Print the diode values as described in the chapter Maintenance check the installed filter and its program in the setup if necessary the filter and the lamp block must be replaced call a service technician LAMP HIGH MINIMUM DETECTION During a calibration measurement the maximum darkness value has been exceeded This can either be due to outside light open device or to dis adjustments on the ADC board amplifier offset Print the diode values as described in the chapter Maintenance and call a service technician LAMP LOW CALIBRATION FILTER The values obtained during calibration filter measurements in the exte
93. sion of Evaluation Results 10 4 10 TD Test directory transmission of test directory Command string to the Reader STX TD ETX start of text key TD end of text The answer string to the computer indicates the test identification number and the test name for each test in the directory Example STX 1 HIV 2 CR Start of text 1st test 2 HERPELISA CR 2nd test 3 RUBELLA CR 3rd test HIV SCREEN CR 4th test Cxx ETX Checksum end of text 10 4 11 TR Test run plate measurement with predefined test operating manual Command string to the Reader example STX TR 5 100001 ETX start of text key TR test identification number plate identification number optional according to directories end of text 68 anthos ht1 ht2 ht3 If the plate identification number is specified it must be different from the numbers already programmed in the directory since if the numbers are identical a no acknowledgement signal will be transmitted to the computer If no particular plate identification number is specified the last ID number is automatically incremented A measurement is started and the raw data evaluated in accordance with the programmed test The measurement and evaluation data are transmitted in the answer string to the computer Refer to the chapter Transmission of Evaluation Results 10 4 12 H Shaking Command string to the Reader example STX H 2 5 ETX start of text key H shaking mode
94. sity OD is determined according to the following formula 83 anthos ht1 ht2 ht3 12 2 operating manual E OD c d e c the concentration of the sample to be measured d the layer thickness of the sample to be measured e the molar extinction coefficient OPTICAL DENSITY VERSUS TRANSMITTANCE OPTICAL DENSITY VERSUS TRANSMITTANCE 100 10 20 a 20 s 70 T B0 ex 0 5 40 4 20 3 20 2 I 10 1 o _ o o as 1 15 2 25 3 35 4 1 11 12 13 14 15 16 17 18 19 OPTICAL DENSITY OPTICAL DENSITY OPTICAL DENSITY VERSUS TRANSMITTANCE bila DENI VERSLS THA NET RANGE 108 010 2 0 09 28 0 08 one 007 ppm 005 IR 005 En 004 ope 003 u 0 02 oe ont Bes 28 000 2 21 22 23 24 25 28 ar 28 28 3 3 33 31 32 34 35 38 ar OPTICAL DENSITY OPTICAL DENSITY Measurement at specific wavelengths Each substance to be measured has a specific transmission profile that indicates its concentration and composition A measurement with white light renders different sample concentrations in a certain spectral range only to a small amount and accordingly inaccurate A higher significance can be obtained by using only that part of the light spectrum which is relevant to prove the respective wavelengt
95. spill fluids in or on the equipment Wash your hands thoroughly after handling test fluids If the equipment has been in contact with hazardous substances it must be disinfected prior to shipment in accordance with the effective provisions Do not touch the plate during movement of the plate transport risk of injury The equipment may not be opened other than as described in the chapter 11 since it contains live parts danger to life Environmental operating conditions e Flat dry clean and vibration proof working area 44 cm x 34 cm length x width Additional room for cables connections printer etc No direct sunlight Clean ambient air free of corrosive vapors smoke and dust Ambient temperature between 15 C and 40 C for HT3 15 C 22 C in temperature mode e Humidity between 15 and 85 Re packing to prevent damage during transport operating manual The original HT2 packing has been specially designed for this equipment The drop and vibration tests described in the specification cannot be guaranteed with any other packing 12 anthos ht1 ht2 ht3 It is therefore recommended to save the original carton with its foam parts 1 accessories box 2 and plate transport brace 3 1 10 Description of operating elements Front view ht1 ht2 ht3 Main switch Display with keybo
96. splay range of the curve both of the following parameters must be set e OD range the range in MOD by which the test fluid will increase or decrease press CE and enter number using the numerical keyboard e Reaction expected reaction process increasing decreasing Inaccurate inputs have no effect on the quality of the measurement data To start the measurement press F5 START MEASUREMENT For the output channel Display press F5 SELECT SCREEN after the measurement has been performed and choose one of the following screens e Maximum slope the measurements are reduced to the value of the maximum slope of the reaction in mOD min e Raw wells the measurement for each cycle is displayed retrieve measurement for various cycles with cursor e Graphics 8 12 Curves per well displayed in series or rows switch with F2 The OD range and the reaction can be changed subsequently using F4 limits e Zoomed graphics The reaction of a well is enlarged on the screen 22 anthos ht1 ht2 ht3 The data of a kinetic test remains in the Quick Menu memory F3 KINETIC DATA until another kinetic or scan measurement is performed even if the device is switched off in the meantime Switch Output channel to display if kinetic data are to be viewed 4 4 F4 SCAN scan and agglutination measurements operating manual Measurement Filter Result Units Qutrut Channel Select Locations left 1 1 risht 76543210957654
97. ss was successfully concluded in the Reader the results are transmitted to the computer as defined In case of an error an error message is transmitted message key E 10 2 Command keys operating manual v display of software version installed F display of filters installed in setup N normal measurement also possible with several wavelengths 4 D bichromatic measurement measurement and reference wavelengths 5 M transmission of 35 transmission values per well agglutination and scan measurements 6 A agglutination measurement with adjustable number of well positions to be transmitted 0 31 7 K kinetic measurement combined with shaking function 8 PD transmission of plate directory plate identification numbers tests status of individual plates 9 PR transmission of the evaluated results of a specific plate in the plate memory 10 TD transmission of the test directory all tests defined in the test memory 11 TR measurement of a plate according to the parameters specified for a certain test in the Reader evaluation of results and transmission 12 H shaking 13 R repeat of the last data transmission only possible if no other remote control or stand alone command has been executed in the mean time 14 B transfer of the plate identification number via barcode reader ad a 63 anthos ht1 ht2 ht3 15 setup information transfer of type o
98. standards is programmed S1 S2 S3 S4 S5 Concentration Value 0 00001 0 0001 0 001 0 01 0 1 Response Value Si S2 S3 S4 S5 All the standards are put on the plate The measured result for the response values are S1 S2 S3 S4 S5 Response Value 0 3 0 6 0 8 0 9 1 1 A new test is programmed Only standard S3 is physically on the plate and its measurement value will be the correction factor for all other standards S1 S2 S3 S4 S5 Concentration Value 0 00001 0 0001 0 001 0 01 0 1 Response Value 0 3 S3 0 8 0 6 S3 0 8 0 8 S3 0 8 0 9 53 0 8 1 1 S3 0 8 operating manual 34 anthos ht1 ht2 ht3 If the response value of standard S3 should deviate from 0 8 all other standards are corrected Si S2 S3 S4 S5 Response f r 0 32 0 64 0 85 0 96 1 06 erste neue Kurve Response f r 0 27 0 54 0 72 0 81 0 90 zweite neue Kurve Driginal Curve Istnew curve 2nd new curve o 000001 0 0001 0 001 0 01 0 1 5 6 3 Quantitative evaluation Settings for quantitative evaluations such as curve calculations coordinate scaling extrapolation etc of concentr 1 MGML fit method CUBIC SFLIHE x axis scale LINEAR y axis scale LOGARITHMIC Extrarolation 204 Factor conc 3 1 Grarhics gt Rerort gmg select item cursor modify CE A A A
99. such substance Never spill fluids in or on the equipment Wash your hands thoroughly after handling test fluids If equipment has been in contact with hazardous substances it must be disinfected prior to shipment in accordance with the effective provisions Do not touch the plate during movement of the plate transport risk of injury The equipment may not be opened other than as described in chapter 11 since it contains live parts danger of life gt Electrical Voltages dangerous to human life are present in this device Before removing any covers disconnect the device from the power source Ensure that the power cord supplied with the unit is used The power cord may only be inserted in a socket outlet provided with a protective ground earth contact The protective action must not be negated by use of an extension cord without a protective grounding contact Do not replace fuses without first removing the main power cord Ensure that only fuses with the required rated current and of the specified type are used for replacement The use of makeshift fuses and the short circuiting of fuse holders is prohibited When the apparatus is connected to the main power source the opening of the covers or removal of components is likely to expose life parts The device shall be disconnected from all voltage sources before it is opened for adjustment or repair Any adjustment or repair of the opened apparatus under voltage should b
100. th the cursor and the respective function key confirm with ENTER Turn page with F5 NEXT 29 anthos ht1 ht2 ht3 5 5 Plate layout functions Setting parameters for the validity of blanks filling direction replicates and control strips operating manual ETHEM z PLATE AUTO REPLICATES WES Direction VERTICAL Humber of Rerlic 2 wells AG CONTROL STRIPS HO FILLING DIRECTION ee select item cursor modify CE A A A E E e Blank validity Validity of the blank value positions on the plate e Plate Mean value of all individually positioned blanks is automatically deducted from other measurement values e Row 12 Free configuration of the blanks on the entire plate The mean value of all blanks in each row is deducted from each sample of this row e g Setting blanks on plate position AO1 and A08 Mean value of AO1 and A08 A01 A08 2 This mean value is deducted from each sample of this row A02 A03 A04 AOS A06 A07 A09 A10 A11 A12 e Col 8 Free configuration of the blanks on the entire plate The mean value of all blanks in a certain column is deducted from each sample of this column e g Setting blanks on A01 BO1 and D01 Mean value of A01 BO1 and DO1 A01 BO1 DO1 3 This mean value is deducted from each sample of this column C01 E01 FO1 G01 H01 Automatic replicates automatically set replicates valid for all samples blanks co
101. the current transmission status e g data transfer etc does not however indicate the remote controlled status Communication can be discontinued by pressing the STOP key 10 1 Principles of the communication process operating manual All transmissions including the acknowledgement signal begin with ASCII STX start of text hex 02 and end with ETX end of text hex 03 1 Receipt of command a command string defined by a command key with a maximum length of 192 bytes is sent to the Reader 2 Transmission of answer an answer string with checksum Cxx is sent to the computer Either a successful command transmission is acknowledged ack or a transmission error message is displayed nack Transmission errors can have the following reasons The Reader did not recognize the command the parameters were not completely transmitted the parameters were not within the specified range etc Example STX Cxx ETX Positive acknowledgement STX Cxx ETX Negative acknowledgement 3 Execution of process measurements access to Reader data Only if the command received a positive acknowledgement 62 anthos ht1 ht2 ht3 1 Command to Reader 2 Acknowledgement Ce 3 Process Execution measurements 4 Data Tansmission lt _ accessto reader data 4 Transmission of result data After the specified proce
102. the testing regulation to be defined by the user from the measured value of the samples available on the microplate Since this value greatly depends on the type of plate used a blank cell is placed at a position on the micro titration plate to be specified by the user and measured together with all other samples in one operation Warming up time Ready for operation immediately after switching on and automatic initialization ready for measurement after 10 minutes pre warming time ht3 30 minutes after switching on in temperated mode otherwise like ht1 and ht2 e Operating voltage 90 130V and 180 260V system voltage Ambient temperature 15 C 40 C operation non temperated mode 22 C 1 C operation temperated mode 25 C 50 C storage e Relative humidity 15 85 non condensing operation lt 95 non condensing storage e Air pressure tolerance 54 000 106 000 Pascal e Height over NN operation to 5000 m 1 3 6 Limits of error and tolerance limits e Wavelength instability A 2 nm filter e Spectral width at half maximum intensity 12 nm at 3450 nm 15 nm at lt 450 nm e Integral portion of misradiation blocking measured with all installed filters and band stop filter RG850 3mm Result is outside of measurement range e Accuracy at 510 nm measuring device Schott NG3 1mm 1 and 0 005 O D at 1 O D e Linearity at 620 or 550 540 492 510 measuring device Grey Scale NG4
103. tion input 140VA max at 230V operating voltage e Adjustable operating voltage 90 130V 180 250V e Mains frequency 47 63 Hz Data connection e Data interfaces Serial RS232 V24 9 pole plug parallel Centronics printer interface 25 pole socket e User interface keyboard with function keys 16 line LCD display for 40 characters per line in the normal text mode 240 x 128 pixels with graphics capabilities operating manual 10 anthos ht1 ht2 ht3 1 4 5 Automation Remote controlled via RS232 1 4 6 Supplementary equipment 1 5 Cleaning Barcode spacing scanner optional Printer all EPSON ESC P or IBM compatible parallel 9 and 24 matrix printers with parallel interface Transfer rate of reader selectable between 300 and 19200 baud Cleaning for all accessible housing parts we recommend cleaning with a disinfecting agent commonly used in laboratories effective against hepatitis B or a 70 alcohol solution Decontamination If the equipment has been in contact with infectious substances it may be disinfected with 37 formaldehyde steam at a temperature of 40 C 4 for 6 to 8 hours in an air volume of approximately 160L and a relative humidity of 85 95 The filters may be disinfected with a 96 ethyl alcohol solution and optical paper by means of short manual cleaning Note Warnings regarding the use of toxic or inflammable ligids page 6 Take care on proper ventilation 1 6 Photometry Monochromat
104. tion takes place the test is invalid ERROR QUANTITATIVE EVALUATION MORE STANDARD POINTS REQUIRED The minimum number of standard points depends on the selected curve fit mode If not enough points were defined in the test definition or if plate evaluation points are lost rejected overflow auto eliminated the curve can no longer be evaluated Possible remedy repeat the test or select another curve fit mode also refer to the chapter Report amp Evaluation ERROR QUANTITATIVE EVALUATION INVALID CURVE DATA A standard curve calculation is no longer possible 59 anthos ht1 ht2 ht3 operating manual Neither standard curves nor concentration values can be obtained Find error by means of the printout ERROR QUANTITATIVE EVALUATION INVALID POINT SEQUENCE The points of the standard curve are not available in an uninterrupted ascending or descending order No concentration calculation is performed Find error by means of the printout Try to re evaluate after eliminating the standard outlier or re evaluation after defining the test without including the faulty standard in the curve calculation ERROR TEST INVALID AUTO REPLICATE ELIMINATION Due to the auto elimination the minimum number of replicates for at least one standard one control or one blank was not attained The test is considered invalid although an attempt to evaluate was made ERROR TEST INVALID CONTROL VALIDATION The test becomes i
105. to its original test F5 Clear To erase individual plates or the entire plate memory Note Make sure printouts are available before starting to erase operating manual Move cursor to the plate to be erased press F5 CLEAR To erase a preselected plate F1 CLEAR ID to erase the entire memory F3 CLEAR STORAGE F5 EXIT to abort 52 anthos ht1 ht2 ht3 9 Error messages 9 1 Error messages in the display A operating manual ABORTED BY USER STOP KEY PRESSED is displayed when a measurement was aborted by pressing the STOP key ABORTED BY USER REMOTE CONTROL lt STOP gt is displayed when a measurement was aborted by an external command A D CONVERTER TIMEOUT The AD converter or one of its driver modules on the ADC Board is defective Call a service technician A D CONVERTER TOO SLOW The AD converter or one of its driver modules on the ADC Board is defective Call a service technician ALL DATA IN PLATE STORAGE WILL BE LOST is displayed when the entire plate memory is to be erased PLATES CLEAR CLEAR STORAGE Erase entire memory irreversibly by pressing F2 CONTINUE or return to the plate directory by pressing F4 EXIT AUTO REPLICATES AND AG CONTROL STRIPS CANNOT BE COMBINED is displayed if both functions were selected in the test program or test edit Plate calculation must be performed by means of an automatic layout function whereas the second function can be simulated manually in the
106. to the Main Menu by pressing STOP or F5 EXIT e Press in sequence F5 SETUP and F1 TEST e Press F4 DIR to make a printout of the test directory e Press F3 PRINT to make a printout of the parameters of the first test e Mark with cursor the second test and repeat procedure e Press in sequence F5 CLEAR and F3 CLEAR STORAGE to delete all tests e After having performed a printout of all tests press STOP to go back to the Main menu e Press F4 PLATES and F2 REPORT A printout of the first stored plate is performed e Mark with cursor the next plate and repeat procedure After having performed a printout of all stored plates Press F5 CLEAR and F3 CLEAR STORAGE to delete all plates e Loosen screw A on rear of lamp casing see drawing e Slide lamp block cover forwards and lift off Lay sideways on the outer casing without unplugging e Loosen tape and remove batteries from their holder e Insert three new 1 5 V alkaline batteries The holder is marked for the direction of inserting the batteries Note Incorrectly inserted batteries will cause storage problems and result in a loss of data e Put lamp cover into place and fasten with screw and corresponding plastic washer To check Program new setup and several tests Note corresponding data or print test parameters Turn device off Check after approx 24 hours If the data does not conform check if the batteries are inserted in the right direction and if the cable
107. ual wells will be displayed on the screen A printout will be necessary for the mean values of replicates 45 anthos ht1 ht2 ht3 The display of measurement data and plate layout is independent of the type of test RUN TEST Screensavailable according to selected test TI Plate Layout Measurement Results Kinetic Data Reduction gt Quantitative Results H gt Standard Data 2 Standard Curve gt Qualitative Results leise Kinetic Raw Data L_ Kinetic Raw Data Graphics 2 Kin Raw Data Graph Zoom gt Relational Results 7 1 Measurement Results operating manual Press F5 SELECT SCREEN move cursor to Meas Result press ENTER Only half of a plate can be displayed on the screen at one time Switch to the other plate half with the cursors For kinetic measurements the values for the programmed data reduction will be displayed instead of measurement results in OD TETMIFOMD Function keys Fi Report Printout of measurement data evaluation results and plate layouts This function is also available in other program sections F2 Transmit Transmission of measurement data to an external computer This function is also available in other program sections F3 Reject Deleting measurement values that are not to be included in the calculation can be revoked Press CE to display a plate matrix on the screen Move the cursors to select the well whose measurement value is t
108. urement filter press CE and move cursor to desired wavelength ENTER Reference filter select reference filter as above if no reference filter is required press NONE Measurement range set measurement range basic setting normal mode Note Using extended mode the indication limit is extended from 3 0 OD to 3 5 OD Wavelengths under 400 nm are not permitted Measurement time is doubled operating manual Shaking plate shaking selectable by pressing CE to YES prior to measuring enter time in seconds using keyboard choose shaking intensity from three levels enter menu by pressing CE select low medium high Output channel three output channels available e Display Measuring values of the measurement filter M reference filter R and their difference M R are displayed on the screen printout of difference or transmission of measurement data also possible after measurement e Printer Immediate measurement printout printout format selectable between M R M R output of three values per plate position and Delta OD output of differences for measurement and reference filters only e RS232 Transmission of measurement data to external computer Start the measurement with F5 START MEASUREMENT 20 anthos ht1 ht2 ht3 4 2 F2 MULTI measurement with several wavelengths simultaneously OQuteut channel Deam select item 3 modify CE Selection of tyre of measurement ar m A A e Number of
109. y The default number is the next consecutive number after the last measured plate Press CE and the alphabet will appear on the display screen in 5 rows and 6 columns Select the row with the cursors and the column with the respective function key Enter numbers with the keyboard Confirm the entire identification number with ENTER Last Position If the entire plate has not been allocated set the last used position on the plate for which the measurement values are to be evaluated Press CE and the microtitration plate pattern will appear on the screen Move cursor to the desired position and confirm with ENTER Output Channel Select the output channel after measuring NONE The measurement values are immediately stored in the plate memory with the status meas measured and can be retrieved at any time DISPLAY The data are displayed on the screen They may be printed or transmitted afterwards PRINTER RS232 Output of measurement values via printer or external computer Note Check printer setup in Setup System and turn printer on Place the plate to be measured in the plate transport system Note The plate position A01 must be in the upper left corner The plate will not fit tightly in the transport system until measuring is started and the automatic centering is activated operating manual Press F5 START MEASUREMENT The plate is being measured For the output channel DISPLAY the results of the individ
110. y for filter OK 5 Filter 405nm Energy for filter OK 6 Filter 340nm Energy for filter OK EPROM CHECKSUM IC 12 1B03 IC 15 0EBB IC 13 C9EO IC 16 5E34 IC14 32F4 IC17 9068 Diode Values RA B D E F G H 1 620nm 3532 3480 3357 3524 3355 3143 3452 3294 3524 2 540nm 3541 3501 3463 3542 3386 3229 3548 3361 3603 3 492nm 3455 3470 3475 3507 3330 3218 3509 3326 3511 4 450nm 3530 3470 3358 3501 3363 3142 3599 3302 3424 5 405nm 3291 3270 3416 3463 3394 3138 3471 3267 3498 6 340nm 3143 3075 3274 3258 3041 3034 3286 3060 3220 7 007nm 63 62 60 4095 61 55 57 61 60 8 008nm 4095 4095 4095 4095 4095 4095 4095 4095 4095 Lamp off 1 620nm 21 23 20 23 22 21 23 20 21 2 540nm 22 23 20 23 22 21 23 20 20 3 492nm 21 24 20 23 22 20 22 20 21 4 450nm 20 22 21 23 21 20 20 24 22 5 405nm 21 23 21 23 22 20 22 21 21 6 340nm 60 56 61 59 59 54 56 59 60 7 007nm 59 55 60 60 58 55 59 59 59 8 008nm 58 57 61 59 58 53 58 60 59 Check ADC 1 23 21 20 23 23 20 22 22 23 2 60 56 51 59 59 51 54 56 59 Check Lamp controller 1 3543 3503 3467 3543 3385 3226 3551 3369 3606 2 1070 1058 1054 1053 1023 969 1075 1017 1080 operating manual 82 anthos ht1 ht2 ht3 12 Principles of photometric measurement Light is electromagnetic wave radiation Rays from 100nm to 400nm are defined as the ultraviolet spectrum of light Only rays in the range from 400nm to 780nm are visible for the human s eye rays with longer wavelengths are called infrared Color impressions

Anthos HT-series Plate Reader operating manual

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