Operating Manual
Contents
1. oiochrom Zenyth 340 Microplate Reader User s Manual Biochrom Ltd 22 Cambridge Science Park Cambridge UK CB4 OFJ Tel 44 1223 423723 E Mail enquiries biochrom co uk olochrom Zenyth 340 User s Manual v 1 0 August 2010 2 Table of Contents Safety imomacoO a e ERE 5 a O AAA A I EID PETS DT TEC PED EE IE SAS 7 1 1 SYSTEM Desc o 7 1 1 1 ANA susci rure ren quete ni exuere aya mic lev Ra Rx a ewe FR RR RR WV ER NEN E NA EA 7 1 1 2 Options 7 1 4 3 A ON 7 1 1 4 Scope ot SUPPIV n do nee een 7 1 2 Technical Dala IERREURRHEEREEERHERFEREEHEREEEEFHEEERERFHEREEEERREERERRELEREUERREEREERRHERRERSEHEEEEEREELEEEESHREREEERRRER 7 124 Table of Specifications 8 1 2 2 Measurements and Weight us gear 8 1 2 3 Power Supply nen ann ne ernennen E nee een 8 1 2 24 Sound Pressure Level ii a hear ea 8 1 2 5 Environmental Conditions ccccceceeeceeeeeeeeeee neces aaeeeeeaeeeeesaaaeeeeaaeeeeeeaaeeeeaeaaeeeeaaeeeeseageeeeags 9 1 2 6 Defects and Abnormal Stresses szusi iaria aaae e NAE AAR E AAi 9 V MET D NORTE 9 1 3 InterfacihnB eE E E 10 1 3 1 Zenyth 340r and 340Frt iiie tena e torte tn ete er nane dt tan aa Ra ra Dieux cu e ee 10 2 Start up and System SO atada 11 2 1 Unpacking and Installation ccooocccconnnococnnnonnnonannnononononcnnnnnnnonnnnnnncnnnnnnnonnrnnonannnnnos 11 2 2 Calibration and Status ChecKk oooccconcconcconoconocononononononononanonononanonanonanonan2. 9 Complete a disinfection certificate and make a copy of the certificate po Enclose the disinfection certificate in the reader package and attach the copy to the outside of the package so that it is clearly visible Zenyth 340 User s Manual v 1 0 August 2010 29 olochrom 3 3 Exchanging the fuse In case of malfunction the fuses may be blown They are positioned in the mains plug on the backside of the instrument Remove the fuse holder with a little flat screw driver and exchange the fuses 2A T Two spare fuses are in the accessories box Put the fuse holder back into the mains plug 3 4 Exchanging the air filter 1 Remove the cover of the fan on the backside of the instrument 2 Exchange the fleece 3 Put on the cover again 3 5 Lamp and Interference Filter Exchange 3 5 1 Required tools Screwdriver for recessed head screws Screwdriver flat 2 5mm Cotton buds for filter cleaning Zenyth 340 User s Manual v 1 0 August 2010 30 3 5 2 3 5 3 biochrom Open the upper housing Disconnect the instrument from the power source Open the two screws M3x10 that fix the upper and the lower housing and open the upper housing now you have the lamp and filters directly in front of you Lamp exchange To remove the lamp unscrew the two screws in the coupler terminal that fix the cables labeled with 1 Remove the distance that fix the complete lamp block labeled with2 Take out the o
3. A CAUTION calls attention to a condition or possible situation that could damage or destroy the product or the operator s work Electrical Safety To prevent electrically related injuries and property damage properly inspect all electrical equipment prior to use and immediately report any electrical deficiencies Contact an Anthos service representative for any servicing of equipment requiring the removal of covers or panels High Voltage This symbol indicates the potential of an electrical shock hazard existing from a high voltage source and that all safety instructions should be read and understood before proceeding with the installation maintenance and servicing of all modules 1 Voltages dangerous to human life are present in this device Before removing any covers disconnect the device from the power source 2 Ensure that the power cord supplied with the unit is used 3 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 4 Donot 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 5 When the apparatus is connected to the main power source the opening of the covers or
4. Chg changes the name of the plate lt enter gt confirm name selection of new plate Name Esc Sel This screen is used to enter plate name of new plate Useable keys Esc leave the menu Sel Selects the letter or number scroll through alphabet and numbers lt enter gt confirm selection after whole name is edited PlateFormat lt lt esc This screen is used to select plate format 6 12 24 48 96 384 well plate Useable keys Esc leave the menu toggle between the different plate formats 6 well 12 24 48 96 384 lt lt back to previous menu item lt enter gt Confirm selection Zenyth 340 User s Manual v 1 0 August 2010 20 olochrom In the diagram all parameters for a new plate definition are displayed In the following section documentation is shown how to define a new plate 1 2 3 AO Y lores AO X A Well diameter 1 O0 O Al Pos X lt lt Esc 0 01mm This screen is used to Define X position of A1 well on plate in mm Useable keys Esc leave the menu Decrements Increments the position lt lt back to previous menu item lt enter gt Confirm selection Al Pos Y lt lt Esc 0 01mm This screen is used to Define Y position of A1 well on plate in mm Useable keys Esc leave the menu Decrements Increments the position lt lt back to previous menu item lt enter gt Confirm selection Delta X Zenytthy lt lt
5. ESC August 2010 21 olochrom This screen is used to define the difference between the center of wells Al to B1 in mm Useable keys Esc Leaves the menu Decrements Increments the position lt lt back to previous menu item lt enter gt Confirm selection Delta Y lt lt Esc This screen is used to define the difference between the center of wells A1 to A2 in mm Useable keys Esc Leaves the menu Decrements Increments the position lt lt back to previous menu item lt enter gt Confirm selection Well Diameter lt lt Esc 0 1mm This screen is used to Define diameter of well Useable keys Esc Leaves the menu Decrements Increments the position lt lt back to previous menu item lt enter gt Confirm selection and store values in flash 2 3 5 Service Menu 2 3 5 1 Adjust Lamp Zenyth 340 User s Manual v 1 0 August 2010 22 olochrom This screen is the main screen of service Useable keys lt lt gt gt Select lt lt or gt gt to change to main screen lt gt lt gt Use these keys to choose service items lt enter gt Use this key to select service items This screen is used to adjust all installed filters automatically If this menu is chosen the filter adjust procedure starts and all filters will be adjusted On display the current filter being adjusted is shown Useable keys lt F1 gt lt F2 gt select adjust lamp YES
6. or not NO lt gt lt gt not used lt enter gt not used 2 3 5 2 Reset count Reset Count No YES This screen is used to reset cycle count After 10000 measurements the user is told to Do Service After the user confirmed the warning the warning will be repeated every 100 cycles until the user or service resets count Useable keys lt F1 gt lt F2 gt reset YES do not reset No x not used enter not used 2 3 5 3 Key click Keyclick ESC On Zenyth 340 User s Manual v 1 0 August 2010 23 olochrom This screen is used to turn key click on or off Useable keys lt F1 gt lt F2 gt used to go back to service main menu lt gt lt gt used to change between on or off lt enter gt confirm selection 2 3 5 4 Auto calibration Autocalibration No YES This screen is used to confirm auto calibration In this procedure zero position for plate and fork transport are calculated and measured Afterwards the new parameters are written to eeprom Useable keys lt F1 gt lt F2 gt Yes No lt gt lt gt not used lt enter gt Use this key to change to main menu If calibration was ok it is displayed on screen Press any key to continue afterwards 2 3 5 5 Change Air filter AirFilt changed No YES This screen is used to reset air cycle count After 500 hours the user is told to change air filter After the user confirmed the warning the warning wil
7. 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 Zenyth 340 User s Manual v 1 0 August 2010 4 olochrom 6 Any adjustment or repair of the opened apparatus under voltage should be avoided but if necessary it must be carried out by qualified service personnel who are aware of the hazards involved 7 Use the equipment only in the intended manner and as specified by the manufacturer otherwise the protection provided by the equipment may be impaired 8 Only the IBM mouse model No 1233618 is tested for EMC compliance and is recommended to be used with the instrument High voltage peaks in the mains supply may disable the function of a computer mouse connected to the reader Unplug and re plug the mouse and eventually re boot the instrument for resuming its function Use a voltage peak filter between power cord and mains socket if repeated failures of the mouse indicate the occurrence of voltage peaks in the mains supply Chemical and Biological Safety Normal operation of the Zenyth 340 Absorbance Detectors may involve the use of materials that are toxic inflammable infectious or otherwise biologically harmful When using such materials observe the following precautions 1 Handle infectious samples according to good laboratory procedures and methods to prevent the spread of disease Wear protective gloves 2 Obs
8. 25 4 Maintenance 28 3 1 Electrical Safety Considerati0ON coooccccocncccocncononcnnncnnonononnnnnnnncnnnoncnnnnrnnnnnnnnnnrnnnnnnnnnnns 28 3 2 Cleaning and DisinfectiOn iia 28 A ET 29 Zenyth 340 User s Manual v 1 0 August 2010 2 ASTM A A 29 3 3 Exchanging the fuse ise ee 30 3 4 E eee ee RENNES 30 3 5 lramprandiliTenrerencelisiiteniExchangemeeemtRttRRRRHWRMRM A 30 So NRedquitedstoolsseeee eere A e TREE 30 S OS A ERES 31 SEN ETF TE 31 Sa 9SFilter Installationsee mee 32 3547 Zenyt340r TE Ss iere ee negetur dee e aere odia 32 3542 Using ADAP ur asien ee 32 4 Appendix re 33 4 1 Principles of Photometry Measurement uuessssnenennenennennennenennnnennenennennnnnenenne nennen 33 44 1 Absorbance Measurements orestes insinua ea at 33 4 12 Measurement at Specific WavelengthS ooooooocococococonononononononononcnnnnnnnonononononnnnnnnnnnncnnnnnnnnnnanana 34 4 2 Evaluation applications cccccescccseeecceececececseueececesseueeseaeesseeessueesseeesseeeessaeessages 35 4 2 1 Quick reference for test programming seeesesesssssssssesssssesese nere rennen nennen rrr rer r nnne rene 35 4 2 1 1 Bichromatic reading sense nennen dann a han anna arena 35 42 1 2 Blankagainst Ally ee en ran area 35 42 1 3 Shaking Funciona alo iio eub fente 35 AQAA Plate Layout cc 35 4 2 1 5 Blank Subtraction scisso inniinn e a eaa aaa ENA a un 35 4 2 1 6 Single Point C
9. 6 Single Point Calibration Index Calculation B Bo Calculation The Transformation Formula allows calculating a ratio to a particular control for all other controls standards and samples Acommon application is the calculation of the percentage of binding with competitive assays e g X X C1 100 where C1 is the maximum binding control With the use of additional concentration factors and correction factors this formula provides also single point calibration and cut off ratios On the printout the result of this formula is reported with its given name 4 2 1 7 Use of Variables Lot dependent values such as calibrator concentrations or correction factors may be programmed as variables Enter default values for the number of variables required During programming you can use the names V1 V2 etc which refer to the list of defined variables Zenyth 340 User s Manual v 1 0 August 2010 35 olochrom At the beginning of each measurement the variables are displayed for review At this time permanent changes can be made so that the values are kept according to current kit lots 4 2 1 8 Quantitative Evaluation Concentrations are calculated by means of the calibration curve Linear regression is specifically applicable if the calibration curve is really expected to be straight on a linear or logarithmic concentration axis Point to Point and Cubic Spline curve fits are generally suited for non linear curves However the C
10. Standard Curve Graph Kinetic reading for extended range ELISA s With photometry endpoint readings the logarithmic scale of optical density may be a limiting factor for some ELISA applications Quantification of TSH for instance would benefit from a broader dynamic range than normally available If the substrate is TMB then the measurement range of the assay can be extended by kinetic reading The result of the kinetic data reduction e g mOD min may be used as the basis for any further quantitative and or qualitative evaluation The maximum slope maximum reaction speed of the color development at 620 nm corresponds to the amount of bound enzyme In fact stopping of the color reaction means setting of an artificial endpoint and uses also the different reaction speed of different enzyme concentrations Example of Test Parameters Measurement filter 620 nm Interval time 20 seconds Number of readings 30 up to 40 Interval shaking 3 5 seconds medium speed Data reduction Maximum slope Most probably the total kinetic measurement time can be set shorter than the normal substrate incubation time TMB measured in the blue phase at 620nm has about one third of the optical density than the same concentration measured after stopping in the yellow phase Samples and standards which would reach overflow in an endpoint reading will remain within the good measurement range during kinetic readings Therefore the dynamic range and
11. source not found Error Reference source not found and to the ADAP user manual Parallel Interface to connect a printer Note All printers which have PCL transmission protocol can be used PS2 port to link mouse or keyboard or both 2 USB ports for connection to an external floppy drive or memory stick VGA port to link to a PC screen LAN port for connecting to a network See chapter 5 2 USB Ports Parallel Port Serial Port VGA port LAN Port PS2 Port Zenyth 340 User s Manual v 1 0 August 2010 10 biochrom Start up and System Setup 2 1 Unpacking and Installation The original Zenyth 340 packing has been especially designed to protect this equipment during transportation It is therefore recommended to keep the original carton with its foam parts and the accessories box for re use in case of future shipments Warranty claims are void if transport damages are caused by improper packing Unpacking of Instrument Check the box for any visible damage during transportation In case of damage inform your supplier immediately and keep the damaged packing Place the device on a suitable working surface Connect the power cable Connect the serial cable to the PC used for Switch on main switch rear left side The Anthos Zenyth 340 performs the complete initialization and shows the main menu after approx 30 seconds 2 2 Calibration and Status Check Before the first use of the instrument after shipment a few automate
12. OM Port select port or Auto Installed Plates cent El Sense COST6 J GRE In Instrument Type select Zenyth in NUNC12 Serial Number 6 6 12 24 COST24 Type the drop down menus 48 48 COST48 NUNCA8 m Temperature Firmware Version V1 0 01 03 2003 Read Temperature l Set Temperature Service Code 1 Type Celsius v OK 3 To measure a plate Go to Reading Quick or the R button in the task bar To confirm that the instrument is connected with the computer select the Read Configuration button The serial number of the instrument should now appear in the Setup gt Instrument dialogue box along with compatible plate types Quick Read Load Plate Eject Plat v Confirm that the correct format and plate type Format Microplate X Plate Type b2 are selected Measurement Postion Al M Sesame y Select All in Measurement Position to read the Measurement Mode Endpoint Photometric y entire pl ate Y Select Endpoint Photometric for basic readings using a measurement and reference filter from In the Quick Read dialogue box Measurement Filter Reference Filter the drop down menu Zenyth 340 User s Manual v 1 0 August 2010 25 olochrom Y Note It is important to use a reference filter to account for optical inference from the plate Place plate with A1 in the upper left corner of the plate transport Select Start Absorbance measurements will appear in the open matrix in ADA
13. P Data can be exported to data analysis software using the Copy icon Data will paste as a matrix with filter wavelength time and date Zenyth 340 User s Manual v 1 0 August 2010 26 olochrom Zenyth 340 User s Manual v 1 0 August 2010 27 blochrom 4 Maintenance ATTENTION Read these pages entirely and carefully All maintenance work requiring opening of the instrument must be performed only with mains disconnected and must be performed by a technical qualified person 3 1 Electrical Safety Consideration See also Safety Information at the beginning of this manual ATTENTION Voltages dangerous to human life are present in this device Read these pages entirely before opening any seal or unscrew the instrument When the instrument has to be opened for a lamp or filter exchange the device must be disconnected from the power source Ensure that only fuses with the required current rate and of the specified type 2 A T are used for replacement The use of makeshift fuses and the short circuiting of fuse holders are prohibited When the instrument is connected to the main power source the opening of covers or removal of components is likely to exposed live parts The device must be disconnected from all voltage sources before it is opened for any exchange or cleaning Any adjustment or repair of the opened instrument under voltage should be avoided but if necessary it must be performed by qualified service
14. accuracy of ELISA s may be significantly improved with kinetic readings A main advantage of kinetic readings is the elimination of the critical timing between adding of substrate and stopping solution for all wells Zenyth 340 User s Manual v 1 0 August 2010 38
15. ading of a standard series In order to correct plate to plate variations of the assay it is highly recommended to have at least one real standard on the plate by which a correction factor is calculated The correction factor is the actual measurement value divided by the initial measurement value of the repeated standard Example A plate with 5 standards is programmed Standard Points S1 S2 S3 S4 S5 Concentration Values 0 1 1 10 100 1000 Response Formulas S1 S2 S3 S4 S5 A new test is programmed The measured OD s of the standards are entered to the variables Standard Points S1 S2 S3 S4 S5 Response Values OD 0 3 0 6 0 8 0 9 1 0 Put to Variables V1 V2 V3 V4 V5 Only standard S3 is used on the plate layout of the new test It is named C1 control and its measured value is used to calculate the correction factor for all other standards Standard Points S1 S2 S3 S4 S5 Concentration Values 0 1 1 10 100 1000 Response Formulas V1 C1 V3 V2 C1 V3 V3 C1 V3 V4 C1 V3 V5 C1 V3 If the actual measurement value of the standard 3 C1 deviates from the original measurement value V3 all standards are re adjusted by the correction factor C1 V3 corrected v 1 0 August 2010 37 4 2 3 olochrom Original Curve Ist new curve OD nd new curve 0 0 00001 0 0001 0 001 0 01 0 1 Fig 5
16. alibration Index Calculation B B Calculation oooocococncnnnnnnnnnnncnononons 35 42 L7 UsSofVarables kissen a E desi A EE E E E 35 4 2 1 8 Quantitative Evaluation missaro nia ie ee 36 4 2 1 9 Set up of Standards and Axis Division ooooococcccccncncncnononononononcnonononcnnnnnnnnnonnnoncnnnnncnnnnons 36 4 2 1 10 Wse of Interpretation declina 36 4 2 1 11 Validati E EE 36 4 2 2 Storing a Standard CUM Cs cceveccccctstatecanevanzcent Conca totae t obe ae anda E RARE Fe gne DRE a RR EK OUR Dew dO 37 4 2 3 Kinetic reading for extended range ELISA s uuununenenenenenenenenenenenenenenenenenenenenenen nennen namen 38 Zenyth 340 User s Manual v 1 0 August 2010 3 olochrom Safety Information All Warnings and Cautions in this document include an exclamation point a lightning bolt or a light burst symbol framed within a triangle Please pay special attention to the specific safety information associated with these symbols Warning and Caution Definitions The exclamation point symbol is an international symbol which serves as a reminder that all safety instructions should be read and understood before installation use maintenance and servicing is attempted When this symbol is displayed in this manual pay special attention to the specific safety information associated with the symbol WARNING A WARNING calls attention to a condition or possible situation that could cause injury to the operator CAUTION
17. apter 2 3 3 2 Install Filters for adding the new filter to the list See chapter 2 3 5 1 Adjust Lamp for lamp adjustment 3 5 4 2 Using ADAP Refer also to the ADAP manual for further information Connect the reader with the PC PC PC cable turn the reader ON and start the software on the PC Enter the menu SETUP READER and click the button READ CONFIGURATION Type in the new filter wavelength and click the button SET FILTER Confirm the next message to adjust the lamp energy of all installed filters with YES Zenyth 340 User s Manual v 1 0 August 2010 32 olochrom 4 Appendix 4 1 Principles of Photometry 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 to the human s eye rays with longer wavelengths are called infrared Color impressions 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 750nm A photometer is an optical electronic measuring device to determine the amount of light absorbed at a specific wavelength 4 1 1 Absorbance Measurements Experimental measurements return the value of transmission T Transmission is defined as the percentage ratio betwee
18. ction of the movement Zenyth 340 User s Manual v 1 0 August 2010 6 olochrom 1 General Information 1 1 System Description Zenyth 340 is a visible and near UV range instrument that uses a tungsten filament halogen lamp and automated wavelength selection via a filter wheel Its mechanical concept is based on Front loading one directional plate transport Fast moving scanning optics This allows flexible adaptation to all plate formats from 6 to 384 wells 1 1 1 Model Versions Zenyth 340r Remote Controlled Filter Photometer Zenyth 340rt Remote Controlled Filter Photometer with Temperature Control 1 1 2 Options Temperature Control ADAP Software 3 modules available ADAP Basic ADAP Plus and ADAP Expert A wide range of Filters 1 1 3 Intended use Zenyth 340 is intended for general laboratory and research use only 1 1 4 Scope of Supply Filters 340 405 450 492 620 Manual Power cable Serial cable to PC Dust cover ADAP BASIC Software Adapter for keyboard and mouse only S model Spare air filters for fan 2 Spare fuses 2 1 2 Technical Data Zenyth 340 User s Manual v 1 0 August 2010 7 1 2 1 Table of Specifications Parameter Specification Measurement Method Transmission Photometer Light Source Tungsten Halogen Lamp Detector Silicon Photodiode Wavelength Range 340 750nm Filter wheel Positions 8 Available Wavelength 340 405 450 492 550 595 620 oth
19. d filter installation is done immediately If wavelength isunchanged the following screen will appear Adjust Lamp ESC No This menu is used whether lamp adjust to specified filter should be done or not Useable keys esc go back to previous filter menu select yes or no lt enter gt confirm selection and adjust lamp for filter V Filterl This menu appears when select yes above V value of Digipoti G value of pre amplifier esc go back to previous filter menu V and G are increased until ADC values reach required value This process can t be interrupted by user Lamp adjust can take up to 30 seconds per filter After the lamp is adjusted the filter adjust procedure for the next filter is started The user has to follow the whole submenu until he has reached filter number 8 After the user pressed enter the data is updated and written to EEPROM Lamp Adjust AJUSTE Zenyth 340 User s Manual v 1 0 August 2010 15 olochrom After lamp is adjusted result of adjustment is displayed If lamp energy is too low so that the energy needed for measurement can t be reached NOK is displayed Check lamp afterwards or optic system Useable keys all keys are used to confirm 2 3 3 3 Set plate SetPlate Esc This screen is used to select standard plate for measurements If HT protocol only 96 well plates are displayed This feature is implemented especially for HT protocol users Useable ke
20. d accordingly inaccurate A higher significance can be obtained by using only that part of the light spectrum which is relevant to prove the respective wavelength 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 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 Zenyth 340 User s Manual v 1 0 August 2010 34 blochrom 4 2 Evaluation applications 4 2 1 Quick reference for test programming 4 2 1 1 Bichromatic reading It is highly recommended to use a reference reading for subtracting uns
21. d re adjustments and a status check shall be performed in the following sequence 2 2 1 Lamp Adjustment The brightness of the halogen bulb is adjusted for each filter installed For Zenyth 340r rt refer to chapter 2 3 5 1 Adjust Lamp Activate Adj Lamp and wait until the automatic procedure is finished Zenyth 340 User s Manual v 1 0 August 2010 11 olochrom 2 2 2 Auto Calibration The movement of the plate transport and the optics transport is adjusted For Zenyth 340r rt refer to chapter 2 3 5 4 Auto calibration Scroll to Auto calibration and activate the function Wait until the automatic procedure is finished 2 2 3 Status Check For Zenyth 340r rt refer to the ADAP software instruction manual Press function key F1 lt Quick Check gt A self test of the instrument is performed which reports the parameters stored after Lamp Adjustment and Auto Calibration Press function key F2 lt Save gt The report file is stored in the selected path 2 3 Zenyth 340r rt remote controlled version 2 3 1 Zenyth 340r rt Menu Structure R E Temperatur E Define E n if plate i Load plate inserted 2 Eje ct Plate 2 3 2 Main Menu Cur XX X Set XX lt lt gt gt EJECT This menu is standard menu if temperature control is installed If temperature control is not installed Zenyth 340 User s Manual v 1 0 August 2010 12 blochrom ANTHOS ZENYTH lt lt gt gt EJECT N
22. e reader with lint free tissues that have been moistened in ethanol or a chlorine solution 4 Put lint free tissues that have been moistened in ethanol or a chlorine solution onto the plate transport mechanism and let it soak for 30 minutes 5 When a chlorine solution has been used carefully wipe off the entire reader with lint free tissues that have been moistened in water 6 Dry the reader by wiping it off with lint free tissues 3 2 2 Disinfection Before the reader is returned to the distributor it must be disinfected and a disinfection certificate must be completed The following procedure must be used for disinfecting the reader 1 Switch off the reader and disconnect it from the mains power supply 2 When used disconnect the reader from any accessories printer and PC 3 Carefully wipe off the entire reader with lint free tissues that have been moistened in a protein degrading mild detergent or a saline solution 4 Carefully wipe off the entire reader with lint free tissues that have been moistened in ethanol or a chlorine solution 5 Put lint free tissues that have been moistened in ethanol or a chlorine solution onto the plate transport mechanism and let it soak for 30 minutes 6 When a chlorine solution has been used carefully wipe off the entire reader with lint free tissues that have been moistened in water 7 Dry the reader by wiping it off with lint free tissues Pack the reader in its original packaging
23. er wavelengths on request Resolution 0 001 OD 0 1m OD in 0 1 OD Measurement Range 0 3 0 OD specified Indication Range 0 4 00D Accuracy lt 1 5 at 1 OD Linearity lt 0 5 and 0 005 OD from 0 1 up to 1 5 OD lt 0 75 from 1 5 to 3 0 OD 400 750nm lt 0 7 and 0 005 OD from 0 1 up to 2 0 OD 340 399nm Reproducibility lt 0 3 and 0 005 OD at 1 000 OD 400 750nm lt 0 5 and 0 005 OD at 2 000 OD 400 750nm lt 0 5 and 0 005 OD at 1 000 OD 340 399nm Measurement Speed for 96 Well Plate single wavelength 10 Seconds dual wavelength 20 Seconds Wavelength Interval 10 Seconds kinetic dual Wavelength Interval kinetic single 20 Seconds Assumed Workload 5000 Measurements Year Temperature Control from 4 C above Ambient up to 45 C in 1 C Steps Measurement Modes single wavelength dual wavelength multiple wavelength scan measurement kinetic measurement olochrom Shaking three speeds 1 2 2 Measurements and Weight Weight 14 4 kg Dimensions width x height x length 47 3 cm x 25 0 cm x 39 8 cm 1 2 3 Power Supply Operating voltage 100 240 V frequency range 50 60 HFuses 250VAC 2 A T 5x20 mm 1 2 4 Sound Pressure Level Maximum sound pressure 71dBA Maximum sound pressure at one meter 62dBA Zenyth 340 User s Manual v 1 0 August 2010 8 olochrom 1 2 5 Environmental Conditions The working area has to be fla
24. erve all cautionary information printed on the original solutions containers prior to their use 3 Dispose of all waste solutions according to your facility s waste disposal procedures 4 Operate the Zenyth 340 Absorbance Detectors in accordance with the instructions outlined in this manual and take all the necessary precautions when using pathological toxic or radioactive materials 5 Splashing of liquids may occur therefore take appropriate safety precautions such as using safety glasses and wearing protective clothing when working with potentially hazardous liquids 6 Use an appropriately contained environment when using hazardous materials 7 Observe the appropriate cautionary procedures as defined by your safety officer when using flammable solvents in or near a powered up instrument 8 Observe the appropriate cautionary procedures as defined by your safety officer when using toxic pathological or radioactive materials 9 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 Moving Parts Do not touch the plate during movement of the plate transport risk of injury Make sure that the lid is always closed prior to a measurement Zenyth 340 User s Manual v 1 0 August 2010 5 olochrom Keep the Zenyth 340 Absorbance Detectors work area clear to prevent obstru
25. l be repeated every 10 hours until the user or service resets count The default interval is adjustable in eeprom Useable keys lt F1 gt lt F2 gt reset YES do not reset No 2 3 5 6 Service password This area is restricted to service technician and is therefore protected with password Service lt lt gt gt Serv Pwd Zenyth 340 User s Manual v 1 0 August 2010 24 olochrom 3 Quick Start Guide 1 To turn on the instrument Connect instrument to a power source using the appropriate power cord Y Check user s manual for important safety information 2 To connect the instrument to a PC Connect to a PC via serial port to serial port or a serial to USB port adaptor Determine the communication port com used by the instrument In the Start menu of the PC go to Control Panel System Hardware Device Manager Ports 3 Toconnect instrument to ADAP software Insert CD supplied with the instrument into PC install ADAP Open ADAP ADAP will prompt for a user ID and password Use the pre set ID and passwords are sadmin sadmin Once logged as sadmin set specific user IDs passwords and administrative rights Select Setup gt Instrument in the menu bar A dialogue box will open Instrument File Instrument Functions 1 Functions 2 Down Upload m Mae Under the Instrument tab Position Filter 1 v Baudrate 3600 X Wavelength 620 Uo BES Y In Baudrate select Auto Sense Send to Instrument Mies Y In C
26. ld lamp block and insert the new one by following the steps from behind toward the front 2 2 l rubber band for filter 7 T T FA 5 7 ATTENTION Take care not to touch the bulb of the new lamp The position of the filament is pre adjusted and when the position changes not enough energy is present Re assemble the instrument and plug it to the mains After the new lamp is installed and the upper housing is fixed again the lamp energy for the filters has to be readjusted See chapter 2 2 1 Lamp Adjustment Zenyth 340 User s Manual v 1 0 August 2010 31 3 5 4 olochrom Filter Installation Next to the filter motor you can see the filter wheel which is surrounded by a rubber band Choose the position where you want to change or add a new filter by turning the filter wheel the numbers of the filter positions are indicated Do not leave a filter position empty between installed filters Pull the rubber band carefully and insert exchange a filter Pay attention that the arrow on the filter frame shows to the right direction of the light The filter must be clean Avoid dust or fingerprints Re assemble the instrument and plug it to the mains After the new filter is installed and the upper housing is fixed again the filter wavelength has to be added to the list of installed filters and the lamp energy for the filters needs to be readjusted in order to enable the new filter 3 5 4 1 Zenyth 340r rt See ch
27. n the total available light energy at the detector measured through air and the residual luminous energy after sample transmission at a specific wavelength T I lo I Light intensity after passing through the sample lo Initial light intensity Transmission has no linear relation to concentration therefore Absorbance Optical Density is calculated from Transmission by the formula A OD log T or A OD log 1 T Absorbance is related in a linear way to layer thickness and sample concentration according to the Beers law The following graph illustrates the relation between absorbance sample concentration and the light transmitted and measured at the detector Absorbance A Molar Absorptivity e pathlength b concentration c Zenyth 340 User s Manual v 1 0 August 2010 33 olochrom Relation OD Transmission E z E 5 a 15 Optical Density It is important to keep in mind that the best measurement range of absorbance is from 0 1 to 1 OD or 90 to 10 of transmission Measurements above 2 OD deal with less than 1 of the original light and will therefore have lower resolution and accuracy 4 1 2 Measurement at Specific Wavelengths A useful physical characteristic of molecules is specific transmission profile which is indicative of its concentration and composition A measurement with white light renders different sample concentrations in a certain spectral range only to a small amount an
28. nnanonanenaninaninos 11 2 2 1 E T To PG DESI ERI 11 222 Autahi bration secca tito eae Eo sco se E Ea eiii 12 223 Status CHECK E 12 2 3 Zenyth 340r rt remote controlled version essseeeseeeeeee nennen 12 2 3 1 Zenyth 340r rt Menu Structure ccccceeeeeeeeeeeeseeseeeseeseeeeseseseeeesseeeeeeeeeseeeeeeeeseeeeeeeeeeeseseeees 12 2 3 2 ETT ia 12 A sais SAEHEIN SEPEBBESCEEFTEITUEEEEEFSEFFELETTLEEEEEFEFFETLETELEFERSSSTEERDEIELEFESEFFLFELLERTEFERTFILTERLETER 13 23341 Str cture of the Setup MENU acciona ic datada 14 2 3 32 Install Filter coincidan rr a ea dial 14 A NS 16 2334 LANE ii ta ee eee es 16 2 3 3 5 SerialsCOMMUMICAEIOM resedit ls A is 16 23 3 6 CHECK Plate soca sicosciueadearcencbausdseadessdcdeaeeaaveddanhdewaevauavasilsidavsge anveadsas docsbaxuaseddlasaideateuaeeddns 17 2337 Temperature Control dis 17 2 3 3 8 NEW Temperatura ande dadas 17 234 Define plate Me Unison iii 19 2 3 4 1 Define plate menu structure uneessnnsnsennnnenennnnnennnnnnnnnnnnnennennenennnnnnnnennnnen nnne nnns nnns 19 2 342 JDefinejplate roter detto de a didas 19 2 3 5 Service MENU octets tee eat 22 23 51 Adjust Lamp wists een 22 2 3 5 2 NT 23 PECSSEC MEME adque 23 2 3 54 Auto callbratiOni i2i eror intecis croce a di 24 235 5 Change All cs Sasse ae 24 2 3 5 6 Service DassWOFd nda da 24 3 Quick Start Guide coimas
29. ote Only at this menu point the Zenyth 340r rt reader can establish a communication with an external PC via serial interface RS232 Useable keys lt lt gt gt Scroll through the menus Eject the plate if plate is not inserted Load plate lt enter gt Eject plate if plate is not inserted Load plate 2 3 3 Setup Menu In the setup menu the following features of the Zenyth 340r rt reader can be performed Filter installation Set Plate Language settings Communication protocol ASTM or HT protocol Plate check Zenyth 340 User s Manual v 1 0 August 2010 13 olochrom 2 3 3 1 Structure of the setup menu w gt gt SetUp l Esc Filter2 Esc Filters lt Exc Esc P Install Filter Filter1 Esc SetP late lt Plate i H Esc Language Language i rH 8 4 Esc ASTM1381 Protocol HT Protocol PlateCheck i 2 3 3 2 Install Filters Install Filter lt lt gt gt Filter This menu is shown when selecting filters Useable keys lt lt gt gt back to setup menu lt enter gt confirm selection and go to submenu filter install Zenyth 340 User s Manual v 1 0 August 2010 14 olochrom Filterl Esc 1 Useable keys lt lt select filter esc go back to install filter menu select wavelength 340 800 nm lt enter gt confirm selection If wavelength is change
30. ow a warning is displayed temperature too low Confirm warning and enter higher temperature Useable keys Esc go back to main menus gt gt change between On or Off Zenyth 340 User s Manual v 1 0 August 2010 17 olochrom change temperature lt enter gt confirm selection Zenyth 340 User s Manual v 1 0 August 2010 18 olochrom 2 3 4 Define plate menu The Anthos Zenyth 340r rt reader can measure the standard plate formats 6 12 24 48 96 384 Additionally the software can manage a tuning of the above mentioned well formats 2 3 4 1 Define plate menu structure Def Plate gt gt __IMIMIMM lt SAS Esc Name lt lt lt lt Al Pos X0 lt A1 Pos YO lt Esc Well Diameter lt lt lt 2 3 4 2 Define plate This menu is used for the definition of different plate types and parameters A plate definition determines the center position of the wells for optimized adaptation of the measurement system to mechanical plate parameters Plates must not have the same plate name A maximum of 90 plates can be defined Def Plate lt lt gt gt Useable keys lt lt gt gt Scroll through the menus Zenyth 340 User s Manual v 1 0 August 2010 19 olochrom Scroll through the Def Plate menu lt enter gt Enter the selected Def Plate menu Name Esc Chg This screen is used to edit new plate name Useable keys Esc leave the menu
31. pecific signals Suited measurement and reference filters are Substrate Measurement Filter Reference Filter PNPP 405 nm 620 nm TMB 450 nm 620 nm OPD 492 nm 620 nm ABTS 405 414 nm 492 nm 4 2 1 2 Blank against Air An automatic self calibration determination of 100 and 0 transmission signals is performed by the reader for each measurement channel and each particular filter previous to readings This is equivalent to the blanking against air which is often mentioned in kit instructions 4 2 1 3 Shaking Function Itis highly recommended to use a short shaking cycle e g 5 seconds at medium speed previous to measurements This will homogenize the color solution color develops at the edges of the well and will produce an equal and stable liquid meniscus in all wells As a result accuracy and reproducibility are significantly improved 4 2 1 4 Plate Layout Singles or replicates of blanks standards controls and samples may be distributed freely across the plate There are no fixed positions used for calculations The software will find the right wells according to the set names in the plate layout The mean value is calculated automatically for all replicates of a blank control standard or sample 4 2 1 5 Blank Subtraction The mean of positions marked as B blank is subtracted automatically from all other wells If a blank should not be subtracted it has to be named as a control 4 2 1
32. personnel who are aware of the hazards involved 3 2 Cleaning and Disinfection All parts of the reader that come into contact with potentially infectious material must be treated as potentially infectious areas The cleaning and disinfection procedures should be performed by authorized trained personnel in a well ventilated room while wearing disposable gloves protective glasses and clothing The following materials are recommended for cleaning and disinfection of the reader e lint free tissues e Protein degrading mild detergent e g Mucasol Rogypon or saline solution 0 9 NaCl e Ethanol 70 or chlorine solution 1500 ppm Never spray directly into the interior of the reader Zenyth 340 User s Manual v 1 0 August 2010 28 olochrom 3 2 1 Cleaning Clean the reader regularly and immediately after spillage This has to be done with due care and attention Always observe laboratory safety rules and regulations Do not use force when cleaning the reader Wipe off spills immediately with soft tissue Avoid sedimentation of dust on the instrument and wipe off visible dust Cover the instrument with the dust cover if not in use The following cleaning procedure should be performed on demand 1 Switch off the reader 2 Carefully wipe off the entire reader with lint free tissues that have been moistened in a mild protein degrading detergent or a saline solution 3 Carefully wipe off the entir
33. plicates of extreme standards into the valid area for results otherwise the mean of replicates is the borderline 4 2 1 9 Set up of Standards and Axis Division Standards have to be entered in increasing order of concentrations always lowest concentration S1 In case of competitive assays this will produce a falling curve If the standard series is produced by a more or less constant adding e g 10 20 30 or even 10 20 40 70 the axis division is linear If there is a multiplication dilution factor e g 2 4 8 16 or 10 33 100 330 the axis division is logarithmic Zero concentration standards must not be used on a logarithmic scale for mathematical reasons If such a standard sequence is suggested in the kit instructions do not use the zero standard for the curve recommended or set it to a small positive value two logs below the second standard e g 0 02 2 4 8 The identifiers for standards on the plate S1 SXL may be replaced by fixed OD values or formulas This allows to use stored standard curves with correction factors refer to 4 2 2 Storing a Standard Curve 4 2 1 10 Use of Interpretation The Interpretation section provides not only cut off calculation based on optical density If the calculation base is set to the result name of the Transformation or to Concentrations also these results may be grouped by means of the cut off formulas Free names can be given to the groups up
34. t dry clean and vibration proof and leave additional room for cables connections computer printer etc The ambient air has to be clean and free of corrosive vapors smoke and dust The instrument is rated to Pollution Degree II and Installation Category II The ambient temperature has to range between 15 C and 40 C for operation and between 25 C and 50 C for storage Indoor use only Humidity has to range between 15 and 75 Height over sea level during operation up to 2000 m 1 2 6 Defects and Abnormal Stresses Whenever it seems likely that safe operating conditions are impaired the instrument shall be made inoperative for example remove the Power Cord etc and shall be secured against any unintended operation Mark the device as unsafe for use This is the case if the device for example e shows signs of visible damage e fails to perform its intended functions e has been subjected to prolonged storage under unfavorable conditions e has been subjected to severe transport stress 1 2 7 Liability In original condition the instrument meets all safety regulations for a risk free operation Biochrom Ltd cannot warrant damages or any resulting costs caused by alterations repairs or modifications of the equipment Zenyth 340 User s Manual v 1 0 August 2010 9 0 Chrom 1 3 Interfacing 1 3 1 Zenyth 340r and 340rt Serial Interface for remote control via ADAP Basic Refer to chapter Error Reference
35. to five 4 2 1 11 Validation The validation formulas describe true conditions on basis of optical density values Zenyth 340 User s Manual v 1 0 August 2010 36 4 2 2 Zenyth 340 User s Manual olochrom A bold warning will be stated on the printout if one of the validation criteria s is not fulfilled Storing a Standard Curve Note The use of stored standard curves is not generally recommended or applicable for the quantitative evaluation of ELISAs 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 S1 for the standard S2 as S2 etc the curve can be copied to another test procedure the in the following way Make a printout of the measured plate with standards containing both concentration values and OD values Copy the original test procedure to a new file by changing the name and editing the test a copy will be stored under the new name Put in the OD values of the measured standards S1 Sx as default values for Variables V1 Vx The layout does not need to contain a real standard anymore Fill the plate just with samples As response values enter the variables V1 Vx instead of the standards S1 Sx When starting a reading with the new test the variables are displayed and may be updated to the last re
36. ubic Spline will not tolerate big jumps and turning points and will show a tendency to swing out at such occasions Point to Point may not be that accurate but is completely save and robust under all conditions For both the concentration axis has to be selected linear or logarithmic according to the sequence of standard points The 4 parameter Fit is a non linear regression especially suited for competitive assay formats Its theory assumes a point of inflection and two asymptotes at the extreme ends The axis division is fixed to lin log Zero concentration standards must not be included in the curve but in case of competitive assays they should be named as negative controls e g NC1 and may serve for the calculation of B B by means of the relational formula X NC1 100 The 4 parameter fit is also generally suited for non linear curves with sandwich assays If a turning point is not present it will adapt to the lower or upper branch of a sigmoid curve It is more stable than the Cubic Spline and will never overshoot but may need slightly longer calculation times Extrapolation set in 96 allows calculating sample concentration also in a small area below and above the extreme standards Extrapolated results are marked on the printout with an X Results out of range are shown with the prefix or to the highest or lowest valid concentration border of extrapolated area A few percent extrapolations are recommended in order to get both re
37. ys Esc go back to setup menu select plate lt enter gt confirm selection 2 3 3 4 Language Language Esc English This screen is used to change language for Zenyth 340r rt Useable keys esc go back to setup menu select language lt enter gt confirm selection 2 3 3 5 Serial communication Setup lt lt gt gt RS 232 This screen is used to select the serial communication protocol The user can select either HT type communication or ADI type communication If submenu protocol is selected Zenyth August 2010 16 olochrom Useable keys Esc go back to setup menu select communication type lt enter gt confirm selection 2 3 3 6 Check plate Plate check Esc This screen is used to activate or deactivate plate check If plate check is activated measurement procedures are stopped if no plate is inserted in device Useable keys Esc go back to setup menu select On Off lt enter gt confirm selection 2 3 3 7 Temperature Control Temperature lt lt gt gt TempCtl This screen is used to set temperature on Zenyth 340rt Useable keys lt lt gt gt scroll through main menus not used lt enter gt confirm selection 2 3 3 8 New Temperature New Temperature Esc Off 30 This screen is used to turn temperature control On or Off and to set temperature The temperature has to be set 4 higher than room temperature If temperature is too l
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