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Varioskan Flash User Manual

1. 39 Figure 2 22 Removing or replacing the adapter for plate w o lid 126 40 Figure 3 23 Transport locks and transport lock tags present cee 44 Figure 3 24 Dispenser sliding cover and measurement chamber door opened A et ene mete De ea er ee a er eae ere 44 Figure 3 25 Front cover removed 0 ceeeccececesesesceteseesesesteseseseeteeseseatenenees 45 Figure 3 26 Transport lock fastened screws 1 4 shown cccc 45 Figure 3 27 Releasing the transport lock screws 1 4 shown 46 Figure 3 28 Transport lock in its horizontal storage position screws 3 and ESTONIA NEEE E EEE Sea uett oad E A clad chee Alea 46 Figure 3 29 Varioskan LumiSens optics transport lock fastened 47 Figure 3 30 Varioskan LumiSens optics transport lock removed 47 Figure 3 31 Tray holder ccceccsesesescessesescesesesescereseseeceneseseecenenees 48 Figure 3 32 Fastening the tray frame to the tray holder 49 Figure 3 33 Close up of the positioning lever when the tray is out 49 Figure 3 34 Varioskan Flash with the dispenser sliding cover open 50 Figure 3 35 Protective cap removed from the dispensing tip 0 40 mm 51 Figure 3 36 Dispenser assembly 00 cece cece csescetesesesteteseseeteteseseatenenees 51 Figure 3 37 Connecting the mains supply Cable 0 0 00 ccc 52 Thermo Scientific Varioskan Flash User Manual 11 Figures 12 Thermo Scientif
2. Accuracy 2 or 0 003 Abs whichever is greater at 200 399 nm 0 2 Abs 1 or 0 003 Abs whichever is greater at 400 1000 nm 0 3 Abs Precision SD lt 0 001 Abs or CV lt 0 5 whichever is greater at 450 nm 0 3 Abs Stray light lt 0 005 at 230 nm Measurement time 10 1000 ms Measurement speed Reads a 96 well plate in 15 s and a 384 well plate in 45 s minimum kinetic interval time from A1 back to A1 Spectral scanning speed lt 2 s well 400 500 nm 1 flash 2 nm steps Plate types 6 384 well plates Thermo Scientific Varioskan Flash User Manual 113 Technical Specifications Performance specifications Table 6 12 Luminometry Performance specifications Luminometry Varioskan LumiSens optics Wavelength selection Filter size Filter wheel with no filter and up to 8 optional filter positions Diameter 24 5 25 4 mm Thickness 3 5 8 0 mm Detector Wavelength range Photomultiplier tube 360 670 nm Sensitivity Dynamic range lt 7 amol ATP well gt 7 decades using flash ATP reaction white 384 well plate Crosstalk lt 1 white 384 well plate Measurement time 10 10 000 ms Measurement speed Reads a 96 well plate in 15 s a 384 well plate in 45 s and a 1536 well plate in 135 s minimum kinetic interval time from A1 back to A1 Plate types 6 1536 well plates Scanning optics
3. Thermo Scientific Varioskan Flash User Manual Rev 1 2 Cat no NO6354 Copyright Copyright 2008 Thermo Fisher Scientific All rights reserved First edition published in 2006 Printed in Finland Reproduction of the accompanying user documentation in whole or in part is prohibited Patents The Varioskan Flash has an international patent and an international patent pending Trademarks Microtiter SkanIt and Varioskan are registered trademarks of Thermo Fisher Scientific All other trademarks and registered trademarks are the property of their respective holders Disclaimer Thermo Fisher Scientific reserves the right to change its products and services at any time to incorporate technological developments This manual is subject to change without prior notice as part of a continuous product development Although this manual has been prepared with every precaution to ensure accuracy Thermo Fisher Scientific assumes no liability for any errors or omissions nor for any damages resulting from the application or use of this information This manual supersedes all previous editions No liability for consequential damages Thermo Fisher Scientific shall not be liable for any damages whatsoever arising out of the use or inability to use this product Warranty statement Thermo Fisher Scientific Microplate Instrumentation Business products are fully guaranteed against defective parts and materials including def
4. Wavelength selection Double monochromators Detector Photomultiplier tube Wavelength range 270 840 nm Emission bandwidth 12 nm Wavelength setting resolution 1nm Wavelength accuracy 3nm Measurement time 10 10 000 ms Measurement speed Reads a 96 well plate in 15 s a 384 well plate in 45 s and a 1536 well plate in 135 s minimum kinetic interval time from A1 back to A1 Spectral scanning speed lt 2 s well 400 500 nm 1 flash 2 nm steps Plate types 6 384 well plates Table 6 13 Incubator Performance specifications Incubator Incubator warm up time From 25 C to 37 C 5 min Temperature range From ambient 4 C to 45 C at ambient 25 C Setting range From 10 C to 45 C in 0 1 C increments Mean temperature of the wells 0 5 C at 37 C ambient 25 C covered 96 well plate Temperature standard deviation 0 3 C at 37 C ambient 25 C covered 96 well plate Liquid warm up time 114 Thermo Scientific Varioskan Flash User Manual 1 h from 25 C to 37 C covered 96 well plate 200 ul water well Thermo Fisher Scientific Technical Specifications Safety specifications Table 6 14 Shaker Performance specifications Shaker Shaking method Orbital shaking Shaking speed 60 1200 rpm Shaking diameter 1 50 mm Maximum centrifugal force 1G Table 6 15 Dispensers Performance specifications Dispensers Syringe
5. Invitrogen http www invitrogen com Technologies Nanolight http www nanolight com Technology GenoSpectra http www panomics com index cfm Promega http www promega com Sigma Aldrich http www sigmaaldrich com Cambrex http www cambrex com R amp D Systems http www rndsystems com BD Becton Dickinson For more information on fluorometric and TRF labels visit the following websites http www cambrex com http www molecular beacons com http www rndsystems com For information on TEMPCHECK plates visit the following Web sites http www TEMPCHECK nl For more information on luminescence visit the following Web sites http www isbc unibo it International Society for Bio and Chemiluminescence Web site Large selection of articles pictures and Web links about luminescence http www lifesci ucsb edu biolum University of California Santa Barbara Bioluminescence Web site Concentrates on marine bioluminescence biology evolution etc Includes pictures about bioluminescent marine organisms http www promega com default asp Thermo Fisher Scientific Literature Fluorescence intensity Time resolved Thermo Fisher Scientific fluorescence References Literature The world s largest selection of luminescent reagents and kits Also has a 8 8 good selection of articles and literature The section on literature is divided according to the relevant measuremen
6. Clean the Varioskan LumiSens base Contact service if the error persists 84 Varioskan LumiSens reference light failure Contact service 85 Dispensing head is not in the correct position Move the dispensing head into the correct position 86 Wrong plate type Use a suitable plate type Dispensing is only possible with up to 384 well plates 88 Code 100 Varioskan LumiSens base installed measurement module not present Install the Varioskan LumiSens measurement module Table 8 19 Warning codes reported Explanation Unable to comply with the defined lag time Suggested action Unable to comply with the defined well interval 102 Unable to comply with the defined wavelength interval 103 Unable to comply with the defined kinetic interval 104 Unable to comply with the defined group interval 105 The timer referenced in the command is not running anymore Your timing requirement is not met Contact service 106 The lamp lifetime has reached its end Arrange for the replacement of the lamp as soon as convenient Contact service 107 108 Calibration validity has expired The command has no effect Protocol time settings did not allow internal runtime calibration This just informs that a command has been used which has no effect in the current measurement method The synchronization of measurements failed
7. Never touch the surfaces of filters or optical lenses with bare hands Do not wash the electronic parts of the Varioskan LumiSens base when you clean it from below Figure 5 57 Never open any other cover of the Varioskan Flash than the dispenser sliding cover Figure 2 2 or the measurement chamber door Figure 2 2 while the instrument is plugged into a power source Thermo Fisher Scientific Routine Operation Switching on e Never spill fluids in or on the equipment e Do not use the instrument if it appears that it does not function properly e Do not open the measurement chamber door while the instrument is busy when the LED indicator is orange e Do not use plates with dimensions exceeding the top rim of the tray Note that the maximum total height of plates is manufacturer related e Do not use priming vessels higher than the actual plate intended to be used in the assay e Never use any liquids that can cause any precipitation or clotting or that contain any mechanical particles with the automatic dispensers e Do not under any circumstances use formaldehyde e Do not spill any alkalines onto any instrument surfaces to avoid damage of the instrument If needed use suitable protection covering e Do not autoclave any part of this instrument Switching ON The Varioskan Flash is equipped with a power switch ON OFF Figure 2 2 and a three color LED indicator Figure 2 2 When the instrument is switched
8. Then replace the dispenser sliding cover and the front cover Figure 5 51 e Install the transport locks of the tray holder and the Varioskan LumiSens optics Refer to How to refit the transport locks on page 104 e Pack the instrument according to the enclosed packing instructions Thermo Scientific Varioskan Flash User Manual 107 Maintenance Service contracts Service contracts Disposal of the instrument AN 108 Thermo Scientific Varioskan Flash User Manual e Use the original packaging to ensure that no damage will occur to the instrument during shipping Any damage will incur additional labor charges e Enclose a dated and signed Certificate of Decontamination see Appendix B both inside and attached to the outside of the package in which you return your instrument or other items e Enclose the return authorization number RGA given by your local Thermo Fisher Scientific representative e Indicate the fault after you have been in touch with your local Thermo Fisher Scientific representative or the Thermo Fisher Scientific technical service department Refer to General specifications on page 111 for details on storage and transportation temperatures It is recommended to maintain and service the instrument regularly every 12 months on a contract basis by the manufacturer s trained service engineers This will ensure that the product is properly maintained and gives trouble free ser
9. 75 81 136 141 areas 141 aspirate tube assembly 50 96 97 98 132 145 autocalibration 35 66 67 126 146 automatic calibration 35 66 67 automatic runtime calibration 17 66 AutoRange 27 67 68 71 72 bandwidth 26 27 113 127 selector 26 bioluminescence 22 23 134 136 145 146 blind plug 4 62 72 77 bottom reading 15 16 24 28 87 90 112 120 131 BRET 23 145 C calibration 32 55 56 58 66 67 70 71 126 127 128 chemical resistance 61 81 133 chemiluminescence 22 134 136 145 complete dispensing tube assembly 41 50 88 97 98 consumables 2 contact pins 48 control switches 31 78 80 crosstalk 29 64 79 94 128 Thermo Fisher Scientific shield 29 64 79 94 128 dead volume 30 115 decontamination 84 88 89 90 91 102 103 104 107 108 129 139 145 Certificate of 104 107 108 129 139 procedure 84 88 89 90 91 102 103 104 108 129 139 detector 24 113 114 diffraction 25 27 125 145 146 angle 145 grating 25 27 145 146 order filters 25 dispenser 6 15 29 49 76 80 95 131 emptying 31 80 103 107 sliding cover 6 29 44 47 50 62 89 103 104 105 107 syringe 78 88 99 100 101 132 washing 31 80 95 dispensing 76 area 49 84 87 91 base 52 53 54 87 92 head 29 50 51 52 62 72 76 77 78 79 80 92 94 96 98 99 128 head holder 51 52 62 77 78 80 92 94 head posit
10. Appendix B Certificate of Decontamination Name Address Tel Fax Name Serial no A I confirm that the returned items have not been contaminated by body fluids toxic carcinogenic or radioactive materials or any other hazardous materials B I confirm that the returned items have been decontaminated and can be handled without exposing the personnel to health hazards Materials used in the unit Chemicals Biological e Radioactive Specific information about contaminants Decontamination procedure Date and place Signature Name block capitals The signature of a Radiation Safety Officer is also required when the unit has been used with radioactive materials This unit is certified by the undersigned to be free of radioactive contamination Date and place Signature Name block capitals PHOTOCOPIABLE Please include decontaminating solution used Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 139 Certificate of Decontamination 140 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Appendix C Thermo Scientific Varioskan Flash Feedback Form Instrument Thermo Scientific Varioskan Flash O Instrument serial no Software serial no from the Thermo Scientific Skanlt Software for Varioskan Flash installation CD cover PURCHASED BY Company Institute Department Addr
11. Contact service Continued Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 125 Troubleshooting Guide Error and warning codes Cont Code Explanation Suggested action 17 The distance between measurement points is too short for scan Contact service measurement The plate cannot be moved as slowly as the distance requires 18 The sampling time for a single result is too long for scan Use normal measurement instead of scan Use measurement The plate cannot be moved as slowly as the a shorter sampling time sampling time requires 19 The requested plate position is outside the mechanical limits of Check the plate template the tray movements 20 The offset voltage of the temperature measurement electronics is Contact service too high 21 The offset voltage of the A D converter board is too high Contact service 22 The background noise of the A D converter board is too high Contact service 23 Error when checking the A D converter board reference voltage Contact service 24 Calibration of the A D converter board gain steps failed Contact service 25 Analog signal outside measurement range This situation is an Contact service error during startup and autocalibration 26 Flash lamp failure Contact service 27 Attempt to set the instrument serial number when it already has Do not try to set the serial number been s
12. Continued 126 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Troubleshooting Guide Error and warning codes Cont Code Explanation Suggested action 47 This is only reported in response to the factory calibration Contact service 48 Varioskan LumiSens module calibration failed Contact service 49 No tray attached to the instrument The tray carriage will not Attach a tray to the tray holder move unless there is a tray 50 Too high background level The background level is checked Clean off any possible liquid spills inside the during internal calibration measurement chamber Contact service if the error persists 51 The dispenser has failed to initialize properly Contact service 52 The dispenser received an unknown command Contact service 53 Invalid dispenser command operand Contact service 54 Invalid sequence of dispenser commands Contact service 56 Dispenser parameter memory error Contact service 57 Attempt to use a dispenser before it is set up Contact service 59 Dispenser plunger overload If you are trying to dispense a viscose liquid with high speed reduce the dispensing speed Contact service if the error persists 60 Dispenser valve overload Contact service 61 The valve position does not allow moving the plunger Contact service 65 Dispenser command buffer overflow Contact service 66 Attempt to
13. Fixed side supports are located on both the robotic tray and the adapter When the tray comes out the side supports prevent the plate from moving It is recommended to use the tip priming feature for accurate dispensing particularly when the dispensing volumes are small for example 1 to 20 pl For more information on the tip priming feature refer to Dispensing on page 76 The tip priming vessel is an 8 well plate strip 1 x 8 Microtiter Solid Strip Assembly Refer to Table 2 2 and Chapter 9 Ordering Information The robotic tray frame designed for robot compatibility has a cavity for the tip priming vessel located in either of two optional locations on the left side position A Figure 2 21 or in the front position B Figure 2 21 of the robotic tray frame This is due to the space requirements of the optional portrait and landscape orientations of the robotic arms If the robotic access is portrait the tip priming vessel must be located on the left side of the microplate However if the robotic arm accesses the microplate in landscape orientation the tip priming vessel must be located in front of the microplate To change the location of the holder for the tip priming vessel from position A portrait orientation to position B landscape orientation or vice versa Figure 2 21 follow these steps Thermo Fisher Scientific Functional Description Tray options 1 Remove the holder for the tip priming ve
14. LumiSens optics LumiSens module Prime Empty switch ee si transport lock lock knob optional with dispenser Prime Empty switch optional with dispenser Dispenser 3 optional Dispenser 2 optional Dispenser 1 optional LumiSens module Reagent basin Blind plug Male connector of the dispensing tube assembly connected Dispensing head holder Dispensing head optional Dispensing head positioner dispensing head in positioner while dispensing blind plug present when no dispensing Reagent bottle holder Figure 2 5 Varioskan Flash internal views 20 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Measurement techniques Fluorescence intensity Time resolved Thermo Fisher Scientific fluorescence Functional Description Measurement techniques This section describes the relevant measurement techniques including fluorescence intensity FI time resolved fluorescence TRF absorbance Abs and luminescence Fluorescence is the phenomenon in which absorption of excitation light of a given wavelength by a fluorescent molecule is followed by the emission of light at longer wavelengths Fluorescence intensity FI of the emitted light RFU at selected excitation and emission wavelengths is proportional to the concentration of the fluorescent molecule being investigated Fluorescent molecules have two characteristic spectra an excitation spectrum which shows the wavelengt
15. Varioskan LumiSens optics transport lock 106 Maintaining a system loe icc udehielaieneiinenis 107 How to pack for service isco oi 2ict es ioucatacoce seg caecidereccact ta fccabscaces 107 Service Contracts n eea a A AEA A 108 Disposal of the instrument sssssssssssssssssesesssetesrsrsrteerererrrereeseseees 108 Technical Specifications ss sssssscsssssssessssssssssssseseeeseeeeeseateass 111 General specifications sesesesesesesesesesessssrsesesrsrsrsetrerrrrererereeseees 111 Performance specifications esseeseseeeseseeeseseeetesrsrsrttrterererereeseseeee 112 Safety specilica onsite uid thie ee aie a cua teas 115 In conformity with the requirements lt 2 3 cssas ca ssesseteseeerssts 116 Frequently Asked Questions ssssssssssssssssssesssessssesssseessseessseeseneesens 119 Oeics Ae oe pte sta reek Beet OEM sear ae 119 Troubleshooting Guide cscssssssssssessssessesessesseseeateasesessesseateateaeens 125 Error and warning codes ih ee ele dea acct ee eae 125 Service request PIOLOCO lan tet isha au esate ee aca te 129 Certificate of Decontatiimacion hiecaciaseaclesaeteestocuegastgans aaciteass 129 Thermo Scientific Varioskan Flash User Manual 9 Contents Chapter 9 Chapter 10 Appendix A Appendix B Appendix C Appendix D 10 Thermo Scientific Varioskan Flash User Manual Ordering Information ccsccsessssesesseesesseseeseeeesessenteateeenesnesteaee
16. 3 36 fully lowered for 30 minutes 2 After the 30 minute period remove the aspirate tubing Figure 3 36 from the detergent or bleach solution and remove all the fluid from the syringe and tubing into a waste container 3 Prime the dispenser a minimum of 10 cycles with distilled or deionized water Leave the fluid pathways filled for storage Remove the dispensing heads Figure 2 5 from the dispensing head positioners Figure 2 5 and do not let any cleaning fluids enter the measurement chamber Figure 5 52 Use external containers To clean the dispensers Figure 3 34 with weak base and acid follow these steps 1 Prime the dispenser with 0 1 M NaOH and leave the solution in the dispenser for 10 minutes with the syringe Figure 3 36 fully lowered Caution Do not spill any alkalines onto any instrument surfaces to avoid damage of the instrument If needed use suitable protection covering A 2 Flush the dispenser with distilled or deionized water 3 Prime the dispenser with 0 1 M HCL and leave the solution in the dispenser for 10 minutes with the syringe fully lowered 4 After the 10 minute period remove the aspirate tubing Figure 3 36 from the 0 1 M HCI solution and remove all the fluid from the syringe and tubing into a waste container Thermo Scientific Varioskan Flash User Manual 97 Maintenance Periodic maintenance Periodic maintenance If the plunger is stuck A Replacing the
17. 43 Pulling up the filter lift 8 Place the filter wheel into the filter lift and press down the filter into the filter wheel all the way to the bottom Figure 3 44 Use a cloth to avoid touching the filter glass with bare hands Figure 3 44 Pressing down the filter into the filter wheel Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 59 Installation Installation setups 60 Thermo Scientific Varioskan Flash User Manual 9 The filter can be removed from the Varioskan LumiSens module by using the green filter lift in front of the filter installation hole Remember also to remove the filter in SkanIt Software for Varioskan Flash 10 Repeat the installation of all required filters as instructed above 11 When ready close the filter installation hole lid and exit the LumiSens Setup dialog The filter installation is ready Thermo Fisher Scientific Do s and Don ts of the Varioskan Thermo Fisher Scientific Flash Do Chapter 4 Routine Operation The operation of the Varioskan Flash spectral scanning multimode reader is controlled by an external computer and run on SkanIt Software for Varioskan Flash Note Operate the instrument only with software and hardware specifically designed or selected for it Thermo Fisher Scientific assumes no liability for the use of third party software applications A Note It is recommended that the assay includes internal quality controls A This
18. 5 Replace the front cover by inserting it into its slots and close the measurement chamber door and dispensing sliding cover arrows in the opposite direction than in Figure 3 25 To clean the optics that is the photometric lens and the fluorometric bottom reading optics follow these steps Caution Do not touch the optical lenses with bare hands Avoid touching the lens of the photometric window Figure 5 52 and the white reference chip Figure 2 18 with bare hands A Thermo Fisher Scientific How to clean the tray A gt gt o How to clean the reagent basin and dispensing area Thermo Fisher Scientific gt Maintenance Regular and preventive maintenance 1 Soak a lintfree tissue in 70 ethanol 2 Wipe over the lens in a straight movement from inside to out Do not use any circular movements when cleaning the optics 3 Repeat the procedure if the lens is not clean To clean the tray follow the instructions below Caution Keep the instrument tray s Figure 2 19 and Figure 2 20 clean to avoid dust and dirt from entering the measurement chamber Figure 5 52 Clean the tray surface including the tray adapters in use Table 2 1 and Table 2 2 at least once a week using a soft cloth or tissue paper soaked in a mild detergent solution soap solution or 70 ethanol Wipe up spills immediately Do not use formaldehyde a If you have spilt infectious agents on the tray decontaminate accordi
19. 7 17 Tray vs plate specific adapter Tray Adapter Identification Dispensing Incubation number Universal 96 well adapter for plate without lid 2 Universal 96 well adapter for plate with lid 3 Universal 384 well adapter for plate without lid 4 Universal 384 well adapter for plate with lid 5 Universal 1536 well adapter for plate w o lid 65 Universal 6 48 well adapter for plate without lid 80 Universal 6 48 well adapter for plate with lid 48 Robotic Adapter for plate without lid 126 Robotic Adapter removed for plates with lids 127 recommended for 1536 well plate reading Q7 How do I calculate the concentration of samples from photometric readings in a microplate A7 There are two possibilities 1 Establish a standard curve on the plate and determine the concentration of unknowns based on the standard curve 2 Use the pathlength correction feature to bring the results obtained in the short light path microplate into line with those obtained in the 1 cm pathlength cuvette Concentrations are calculated from these corrected absorbances according to Bouguer Lambert Beer s law 120 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Thermo Fisher Scientific Frequently Asked Questions Q amp As Q8 Can I use the pathlength correction calculation with any kind of plate A8 The pathlength correction factors can be used with either a 9
20. Asked Questions Q amp As Q15 What kind of lamp is used A15 A xenon flash lamp Q16 What is a molar absorption coefficient A16 A molar absorption coefficient is a measure of absorptivity or the probability of light absorbing at a particular wavelength for an analyte under specific conditions of pH temperature and solvent Thus a specific amount of material at specified conditions will absorb a specific fraction of light striking it For nucleic acids of unknown base sequence and length the molar absorption coefficient is unknown Universal factors can be used instead derived from a mean value extinction coefficient of DNAs of average base compositions measured in a 1 cm pathlength cuvette The accepted factor is the inverse of the extinction coefficient Note Pathlength correction has to be used when using these universal factors in a microplate A Double stranded DNA factor 50 based on 1 cm pathlength Single stranded DNA ss factor 33 based on 1 cm pathlength RNA factor 40 based on 1 cm pathlength Concentration of a sample of dsDNA ODaz60 50 pg ml QI7 Can I use the Varioskan Flash to measure the expression levels of reporter genes A17 Yes Varioskan Flash can be used to measure the expression level of any reporter gene that has either a photometric or fluorometric detection system available Expression levels can be measured both from intact cells for example green fluorescent proteins and their variants an
21. Check How to pack for service on page 107 You will find instructions on how to proceed before shipping the instrument for service to Thermo Fisher Scientific Oy Check that any necessary decontamination procedure has been carried out before packing Refer to Decontamination procedure on page 102 and Certificate of Decontamination on page 129 Ensure that the Certificate of Decontamination see Appendix B Certificate of Decontamination as well as the return authorization number RGA are sent with the instrument The Thermo Fisher Scientific technical service department will keep you up to date with the progress of service and provide you with any further details you might need for example on maintenance serviceability troubleshooting and replacement Certificate of The decontamination procedure is required before shipping the instrument to Thermo Fisher Scientific Oy for example for repair If Decontamination for any reason the instrument is shipped back to Thermo Fisher Scientific Oy it must be accompanied by a dated and signed Certificate of Decontamination which must be attached to the outside of the package containing the instrument Refer to Appendix B Certificate of Decontamination and Decontamination procedure on page 102 Failure to confirm decontamination will incur additional labor charges or at worst the items will be returned for proper cleaning Before returning any instrument s
22. Figure 3 27 Keep the screws there until needed for future relocation or transportation of the instrument 7 Remove the bottom screw marked number 3 and slightly loosen the bottom screw marked number 4 of the transport lock using the hexagonal screwdriver supplied Figure 3 27 Fasten screws 1 and 2 into their storage site Remove screw 3 and loosen screw 4 Figure 3 27 Releasing the transport lock screws 1 4 shown 8 When you have loosened screw number 4 so that the transport block support moves turn the support into a horizontal storage position towards the interior of the instrument that is to the side of the track mechanism bar Figure 3 28 Fasten screw 4 after having turned the transport lock support into its horizontal storage position Fasten screw 3 back into the same hole from which it was unfastened Figure 3 28 Transport lock in its horizontal storage position screws 3 and 4 shown 9 Tighten the bottom screw marked number 4 firmly once you have turned the transport lock support into its horizontal storage position Figure 3 28 Thermo Fisher Scientific Installation Installation setups 10 Fasten the bottom screw marked number 3 back into the same hole from which it was unfastened Figure 3 28 The transport lock is now released 11 Replace the front cover of the instrument Figure 3 25 and Figure 5 51 and close the dispenser sliding cover and measurement cha
23. Scientific Varioskan Flash User Manual 17 Introduction to the Varioskan Flash Advantages of using Varioskan Flash 18 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Chapter 2 Functional Description Instrument This section shows the front back and internal views of the Varioskan ay out Flash instrument Front view The front view of the Varioskan Flash instrument is shown in Figure 2 2 ONORE gwit lt Dispenser sliding cover pne Power busy and error See Measurement chamber door LED indicator VARIOSKAN FLASH lt _ Front cover fbemne Indentation for lifting Varioskan Flash Figure 2 2 Varioskan Flash front view Back view The back view of the Varioskan Flash instrument is shown in Figure 2 3 and Figure 2 4 Cooling fan outlet USB connector Serial connector ON OFF switch i Mai t aminn ains power supply connector Type label Figure 2 3 Varioskan Flash back view Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 19 Functional Description Instrument layout USB connector iL RS 232 C _____ _ Serial connector eo Mains power supply connector amp SERIAI amp A Figure 2 4 Close up of the computer and mains supply connectors Internal view The internal view of the Varioskan Flash instrument is shown in Figure 2 5 The green color alerts the user to find units parts to be handled
24. Table 9 22 Codes for upgrade Kits cecceccccecscesesteseetesestesestesesteseeeeees 132 Thermo Scientific Varioskan Flash User Manual 13 Tables 14 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Chapter 1 Introduction to the Varioskan Flash Ove FVIG W The Varioskan Flash Figure 1 1 is an advanced spectral scanning multimode reader It is used to measure fluorescence intensity FI time resolved fluorescence TRF absorbance and luminescence in end point kinetic and spectral measurements in the UV Vis NIR range from appropriate microplate formats In fluorometric and luminometric measurements 6 to 1536 well plates can be used and correspondingly 6 to 384 well plates in photometric measurements Incubation can be carried out in a controlled incubation temperature The instrument also allows shaking and reagent dispensing The instrument is run on Thermo Scientific SkanIt Software 2 4 3 or greater for Varioskan Flash which controls all the instrument functions and provides data processing as well as reporting functions The Varioskan Flash is available in the following configurations e Varioskan Flash Cat no 5250030 e Optional Varioskan LumiSens module Cat no 5250500 e Optional one to three dispensers with 1 ml syringe Cat no 5250510 per dispenser e Varioskan Flash including bottom reading Cat no 5250040 e Optional Varioskan LumiSens module Cat no 5250500 e Optional one to
25. The second measurement started later than requested 1 Dark level interpolation buffer overflow Reduce well group size use dark level interpolation between wells or do not use interpolation at all 1 The minimum timing depends on the combination of measurement parameters plate movement parameters dispensing parameters and volume number of wavelengths and number of measured points 128 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Troubleshooting Guide Service request protocol Service request If the Varioskan Flash requires service contact your local Thermo l Fisher Scientific representative or the Thermo Fisher Scientific technical protoco l service department Do not under any circumstances send the instrument for service without any prior contact It is imperative to indicate the fault and nature of the required service This will ensure a faster return of the instrument to the customer Your local Thermo Fisher Scientific representative or distributor will take care of sending a complaint form that is the Warranty Claim Technical Sheet to the Thermo Fisher Scientific technical service department The Warranty Claim Technical Sheet contains a more detailed description of the fault symptom or condition Give all the necessary information to the distributor who will fill out and forward the Warranty Claim Technical Sheet to the Thermo Fisher Scientific technical service department
26. Two different kinds of trays can be used with the Varioskan Flash 1 the universal tray and 2 the robotic tray Refer to Table 2 1 and Table 2 2 The universal tray Figure 2 19 can be used with or without incubation It is suitable for the most common 6 to 1536 well plate formats Fluorescence intensity TRF and luminometric measurements can be carried out with 6 to 1536 well plates and photometric measurements with 6 to 384 well plates You can use a microplate with or without a lid cover depending on the adapters used The adapters are detachable with two thicknesses depending on whether it is intended for microplates with or without lids If you want to dispense use an adapter intended for use with microplates without lids Refer to Universal tray with adapters on page 35 The robotic tray Figure 2 20 is intended for use with robots with 96 to 1536 well plate formats You can use a microplate with or without a lid The robotic tray supports the robotic configuration of either portrait or landscape orientation Dispensing is possible with the elevation adapter intended for 96 and 384 well plates without lids Remove the adapter when you use microplates with lids Refer to Robotic tray on page 37 The tray is inserted into the tray holder Refer to How to install the tray on page 48 The A1 well position is marked on the tray frames to facilitate easy and correct insertion of the microplate Thermo Scientific
27. Varioskan Flash User Manual 33 Functional Description Tray options Tray composition The tray composition is shown in Figure 2 19 and described below Means of identification of tray adapter combinations 384 well adapter for plate without lid 4 Tip priming vessel Positioning lever Screw fix Position calibration hole s Holder for tip priming vessel Universal frame Figure 2 19 Assembly picture of the universal tray Screwfix The tray is fastened to the tray holder which is part of the track mechanism by a screw fix Figure 2 19 Refer to How to install the tray on page 48 Frame The universal frame Figure 2 19 is the basic part of the universal tray into which adapters are fitted Adapter The adapter Figure 2 19 is the detachable part that is fitted into the universal frame There are adapters for microplates with or without lids Some adapters support incubation others do not Table 2 1 and Table 2 2 Means of identification of For automatic identification of tray adapter combinations there are tray adapter combinations codes that refer to the coding system employed Figure 2 19 The adapters are also marked with a visual identification number and the text WITH LID or NO LID to help differentiate them from one another Positioning lever The positioning lever Figure 2 19 is used for automatic positioning of the microplate Holder for tip priming vessel It is recommend
28. Varioskan Flash User Manual Cat no N06355 When you press the Prime control button continuously the instrument fills and then empties the syringe by 20 of the syringe volume for the first ten times after which it fills and empties the syringe to the whole volume as long as the button is continuously pressed The syringe will remain empty when the user stops pressing the button Note that washing and rinsing solutions are not automatically changed but have to be changed manually Dispenser washing can be carried out with the dispensing head located in the dispensing head holder and with a small vessel underneath Caution Carefully insert the dispensing head into the dispensing head holder in one go Otherwise the dispensing head can become damaged or contaminated A Helpful hints To maintain dispenser performance keep the following in mind when operating the dispensing modules e Wipe up all spills immediately 80 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Chemical resistance of Thermo Fisher Scientific the dispensers A gt E Routine Operation Other functions e Dispensing cold fluids may cause leaks due to differing coefficients of expansion of Teflon and glass Leaks may occur when dispensing fluids that are at or below 15 C 61 F e Use organic solvents in the dispensers with caution Using organic solvents may reduce tubing and seal life Refer to Chemical resistance of
29. absorbance log 1 transmittance log hb dimension O D 146 Thermo Scientific Varioskan Flash User Manual Glossary fluorometry order sorting The grating monochromator will transmit multiple wavelengths which are basic wavelengths divided by an integer By employing a suitable optical filter the correct wavelength is selected photometer A device measuring absorbance or optical density in this context photometry The measurement of the properties of light particularly luminous intensity photomultiplier tube PMT A photoelectric cell that converts light into electric current and amplifies the current priming Completely filling the dispenser tubing and syringe with bubble free fluid to allow sustained reproducible dispensing action The air in an unprimed line acts as a spring adversely affecting accuracy and precision quadruple monochromator Dual serially connected double monochromators quantum yield Q The ratio of the number of emitted photons to the number of excited molecules Fluorophores differ in quantum yield the higher the Q value the more fluorescent the compound is The theoretical maximum of Q 1 is for a highly fluorescent compound and Q 0 corresponds to a non fluorescent compound reference chip Reference chip Figure 2 18 present for validation of the fluorometric optics RFU or rfu Relative Fluorescence Fluorometric Units The arbitrary units in which fluorescence intensity i
30. and consist of so called ON shaking periods and OFF times pause periods between shaking periods Figure 4 48 or be a constant shake where the ON time equals the duration or total time of the shake You can also select whether the shaking sequence starts or ends with a shaking ON time Refer to the SkanIt Software for Varioskan Flash User Manual Cat no N06355 Thermo Scientific Varioskan Flash User Manual 73 Routine Operation Other functions OFF ore on of on oF On of ON OFF S e _ON OFF time Duration _ Figure 4 48 ON OFF period time and total shaking time The speed is adjustable from 60 to 1200 spm shakes per minute in 60 rpm increments The maximum diameter and speed depends on the well size the larger the well the smaller the allowed diameter The diameter of the orbital movement is adjustable from 1 to 50 mm Centrifugal forces greater than 1 G are automatically prevented Caution Some combinations of speed and diameter may cause too high centrifugal forces inside the well area resulting in spills inside the measurement chamber Figure 5 52 A The speed and diameter combinations for the different plate formats are presented in Figure 4 49 Only the recommended dark gray speed and diameter combinations should be used Speed and diameter combinations for 6 well plates 1200 1080 960 840 720 600 480 360 240 120 0 0 5 RPM 10 15 20 25 3
31. aspirate tube assembly or the complete dispensing tube assembly 98 Thermo Scientific Varioskan Flash User Manual 5 Prime the dispenser a minimum of 10 cycles with distilled or deionized water There are three parts which require periodic maintenance tubing syringe and valve If they become worn out you are likely to notice these symptoms e Poor precision and accuracy e Air bubbles e Leakage e Drops and spills The frequency of replacement will depend on the duty cycle fluids used and instrument maintenance If any of these symptoms occur and it is not obvious which component is causing the problem it is easiest and most economical to replace one component at a time in the following order 1 dispensing or aspirate tubing that is the input and output tubing Figure 5 60 and or the dispensing tip Figure 5 58 2 syringe Figure 5 60 and 3 valve Figure 5 60 Improper washing of a syringe may cause the plunger to get stuck The following may help 1 Remove the syringe see Replacing a dispenser syringe on page 100 and soak it in alcohol or detergent solution 2 Ifthe plunger does not move after this you need to replace it 3 Ifthe plunger moves rinse the syringe carefully with distilled or deionized water remove the plunger rinse it and allow the syringe and the plunger to dry separately Caution If the dispensers are not properly installed leakage may occur A To remo
32. can be turned off no If selected the dispenser automatically dispenses 1 to 10 pl reagent with the 1 ml syringe and 5 or 10 pl reagent with the 5 ml syringe into the tip priming vessel every time the instrument fills the dispenser syringe provided that the dispenser volume is in the range of 1 to 50 pl This makes the volume of the first well equal to that of the others thus compensating for the so called drawback phenomenon It is recommended to use the tip priming feature to achieve greater accuracy when the dispensing volumes are small Thermo Fisher Scientific Pull back feature Thermo Fisher Scientific Dispensing and measurement Routine Operation Other functions Note Tip priming is a different procedure from manual priming that must be performed when a reagent bottle Figure 3 34 is installed next to the dispenser and the dispenser tubes are completely empty A The tip priming vessel is an 8 well plate strip 1 x 8 Microtiter Solid Strip Assembly There is a cavity for the tip priming vessel in the left side of the universal tray position A Figure 2 19 The robotic tray has two optional locations for the tip priming vessel either position A or in the front of the tray position B Figure 2 21 depending on the robotic integration of either portrait or landscape orientation The 8 well plate strip piece should be changed after about 250 tip primings if the priming volume is 10 pl and after 2500 tip primings if t
33. can frequently provide the service engineer with information that can assist in the diagnosis of problems and minimize the down time An example of a typical user log is presented in Table 5 8 The format of the log can vary to meet the overall requirements of the facility but should include all activity problems abnormal response and any other information that is relevant to the operation of the system Table 5 8 Example of a system log User Date Comments J Smith 12 2 08 Dispensing tip 0 40 mm replaced OK L Davis 14 2 08 Varioskan LumiSens base cleaned OK C Mayo 26 2 08 Dispenser tubing replaced OK J Smith 7 3 08 Annual service OK A blank system log table that can be copied for use is in Appendix A System Log Copy the table as many times as necessary but leave the blank original inside the user manual To pack for service follow the instructions presented below Caution It is important that the instrument is thoroughly decontaminated before it is removed from the laboratory or any servicing is performed on it A When you ship the instrument for service remember to e Inform about the use of hazardous materials e Decontaminate the instrument Empty the dispensers and remove any loose items from the tray for example plates and priming vessels before decontamination e Remove the tray Figure 2 19 and Figure 2 20 and the complete dispensing tube assemblies Figure 5 60 after decontamination
34. for fluorometric measurement and for photometric measurement at the measurement position F Simultaneously the measurement optics collects emission light which is fed to the emission reading channel 26 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Emission reading module Thermo Fisher Scientific Functional Description Optical system Reference sensor Excitation light Emission light Figure 2 9 Measurement optics The excitation beam intensity is measured by the reference sensor before the measurement beam enters the well The reference sensor value is used to correct the result level to compensate for long term and short term flash intensity fluctuations The emission optics Figure 2 10 is basically similar to the excitation optics Refer to Excitation optics on page 25 Warning Do not open the optical covers under any circumstances There is a risk of ultraviolet radiation injury Only authorized service personnel has permission to open the optical covers A Diffraction Photo Emission light 3 Mono 4 Mono order multiplier from the well chromator chromator selection tube filters PMT Figure 2 10 Emission optics Emission monochromators Two diffraction grating monochromators are connected serially as in the excitation optics to gain high stray light rejection The monochromator bandwidth is 12 nm Emission diffraction order filters Emission diffraction order fi
35. immediate damage to the instrument A b gt Pe D How to clean the To clean the measurement chamber follow these steps measurement chamber 1 Switch the Varioskan Flash off by pressing the power switch Figure 2 2 on the left side panel of the instrument into the OFF position 2 Lift up the dispenser sliding cover Figure 3 24 and also open the measurement chamber door Figure 3 25 slightly 3 Ifyou suspect that liquids have entered the measurement chamber Figure 5 52 remove the front cover by lifting it away from both sides according to Figure 5 51 Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 89 Maintenance Regular and preventive maintenance Do not touch the photometric or the bottom reading optics lens with bare hands on the top How to clean the optics in the measurement chamber A 90 Thermo Scientific Varioskan Flash User Manual Lift up the dispenser sliding cover Open the measurement chamber door slightly Lift the front cover away from both sides Figure 5 51 Front cover removed 4 Ifyou have spilt infectious agents into the measurement chamber Figure 5 52 decontaminate according to Decontamination procedure on page 102 Otherwise clean the measurement chamber surface using a soft cloth or tissue paper soaked in a mild detergent solution soap solution or 70 ethanol Figure 5 52 Internal view of the measurement chamber
36. installed and are located on both the right hand Dispenser 1 and Dispenser 3 and the left hand Dispenser 2 side of the dispensing area Figure 3 34 Thermo Scientific Varioskan Flash User Manual 49 Installation Installation setups A Dispenser 2 50 Thermo Scientific Varioskan Flash User Manual Caution If the dispensers are not properly installed leakage may occur A Note that the complete dispensing tube assembly that is the output tubing is always closer to the dispensing positions than the input tubing Dispensers 1 and 3 are alike while Dispenser 2 varies with the input and output tubings being situated vice versa The output and input tubing positions therefore depend on the flow direction of the dispenser that is clearly marked on the dispenser Each dispenser is equipped with an individual electronic dispensing head position sensor Connect the male connector of the dispensing head position sensor to the corresponding female connector on the instrument Dispenser 3 Dispenser 1 Reagent bottle Figure 3 34 Varioskan Flash with the dispenser sliding cover open Note that the aspirate tubing Figure 3 36 is factory installed into the right Dispenser 1 and Dispenser 3 or left Dispenser 2 hole of the valve Ensure that the aspirate tubing is finger tight The aspirate tubing is used to fill the syringe with reagent When using a dispenser make sure the aspiration tube end is completely submerged in
37. measurement o cez jseteeees MiecesiciteccesdisascsarepecxsPteesdiane 70 Photometric spectrum scanning 255s ne ese te att read 71 Luminometric measurement ssesesessesseresesressersrrsresserserseteseeserseeesee 71 Luminometric spectrum scanning ssesessseessersersreeserserseteseeserseresee 73 Serde d layer tise stecnet ana kedisini a N ara S 73 Other fUNCtIONS sisena 73 Orbital shaking isein noniin ti aoee A a orita EE EEE y 73 PTLD ET ee aceasta ed ea ARR 75 DIS POR SIN gennana oiii iia a aaia i aa iiaea 76 Primigenia enan a ea aiga 78 TIP PU MMI Gaia csouieanasectebacineatsnancobsighiectiaiassoeseaneseancaaprnaascuavedeile 78 Pull baek feature especie arcs sapere ia eine etapari 79 Dispensing and measurement s sssessesestetsesistsesisesesesrersesrees 79 Emptying oionn iena e ia aaas 80 Dispenser WAST chen rania a a EE 80 Piel pial Binter a a e ER ieee 80 Chemical resistance of the dispensers scssseeseesereeeeeeneeseeeees 81 Thermo Fisher Scientific Thermo Fisher Scientific Chapter 5 Chapter 6 Chapter 7 Chapter 8 Contents Shutdown nhs 3 cee cece hate eshte tet hoe cance E 84 Emergency situations ninrin eari seri eiii ans 85 Maintenant sicasicssnscicanstandccndeeadsontsentnandatdsapsuuttusncuined ssaustisnniesnapnestutanitankas 87 Maintenance Checkhistap 2i tnc t2 sesh savor late RNA 87 Regular and preventive maintenance lt 5 Aci ea ee eae 88 How to clean the measurement chambef sessesese
38. measurement in the UV Vis NIR wavelength range e Allows optimization of the measurement wavelengths according to the application needs e Allows use of freely selectable wavelengths and spectral scanning e Enables measurement of multiple labels from the same well e Allows optimization of the assays to different plate formats depending on the throughput requirements e Enables precise incubation of temperature critical assays due to the unique design of the universal tray e Has selectable top bottom reading that allows to define the optimal reading position for fluorescence measurements depending on the instrument configuration e Enables fast kinetic measurements due to simultaneous measurement and dispensing e Enables measurement of several measurement locations multipoint within the same well with 6 to 96 well plates to analyze intra well variations of the sample e Has the ability to add reagents in any order or in any phase of the kinetic assay allowing execution of sequential multistep assays e Enables automation due to robot compatibility Thermo Fisher Scientific Introduction to the Varioskan Flash Advantages of using Varioskan Flash e Is controlled by SkanIt Software for Varioskan Flash that provides features required to make comprehensive calculations and reports e Ensures high quality performance due to automatic runtime calibration and operational checks including safety features Thermo Fisher Scientific Thermo
39. monochromators dynamic range Dynamic range refers to the range of signals an instrument can read from the minimum to the maximum detectable For example dynamic range of seven decades means that the difference between the lowest and highest signals that can be measured is 10 emission The release of light from a fluorochrome when an electron falls from an excited state to a lower energy state of the molecule error message Indication that an error has been detected excitation The absorption of light energy by a fluorochrome during which electrons in the fluorochrome molecule are boosted to a higher energy level fluorescein An example of a fluorescent dye emitting green fluorescence fluorescence The emission of light from a fluorochrome the wavelength of the light generally being of longer wavelength than that of the absorbed light fluorescence lifetime The period of time elapsed between when a fluorophore is excited and when it emits light This is between 4 and 10 ns for most standard fluorophores and roughly 1 ps for long lived lanthanides used for TRF measurements Cf T tau fluorochrome fluorophore A molecule or chemical group that emits fluorescence fluorometer Instrument used for measuring the intensity of fluorescent radiation Also known as fluorimeter Thermo Scientific Varioskan Flash User Manual 145 fluorometry The measurement of fluorescence Also known as fluorimetry FRET Fluore
40. not dispense into the instrument by mistake Ensure that a correct microplate has been inserted and that the microplate or tip priming vessel is not too full Insert the dispensing head carefully into the dispensing head holder Otherwise the dispensing head can become damaged or contaminated To avoid stray light use blind plugs to close empty dispensing positions Always remove or empty the tip priming vessel at shutdown When you change the dispensing tip remove the protective cap that protects the sensitive tip Figure 3 35 Exchange the 8 well plate strip piece 1 x 8 Microtiter Solid Strip Assembly that is the tip priming vessel after about 250 tip primings if the priming volume is 10 pl and after 2500 tip primings if the priming volume is 1 pl Check the installation and maintenance checklists Decontaminate the instrument thoroughly before it is removed from the laboratory or any servicing is performed on it Do not use for self testing Do not touch or loosen any screws or parts other than those specifically designated in the instructions Doing so might cause misalignment and will invalidate the instrument warranty Do not open the optical covers under any circumstances There is a risk of ultraviolet radiation injury Only authorized service personnel has permission to open the optical covers Avoid disturbing any of the optical system components including optical covers and the reference chip Figure 2 16
41. or item s ensure that they are fully decontaminated Confirm A or B status Refer to Appendix B Certificate of Decontamination Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 129 Troubleshooting Guide Certificate of Decontamination 130 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Varioskan Flash List of spare parts and accessories Thermo Fisher Scientific Chapter 9 Ordering Information Contact your local Thermo Fisher Scientific representative for ordering and service information Table 9 20 Instrument catalog numbers Code Instrument System 5250030 Varioskan Flash 5250040 Varioskan Flash incl bottom reading 5250500 Varioskan LumiSens option 5250510 Dispenser option w 1 ml syringe 1 2 or 3 Table 9 21 Codes for spare parts and accessories Code Item Quantity N06354 Varioskan Flash User Manual 1 N06355 Skanlt Software for Varioskan Flash User Manual 5187080 Skanlt Software Research Edition 5187090 Skanlt Software Drug Discovery Edition with 21CFR1 1 N03078 Universal tray incl 96 384 and 6 48 well adapters for plate w o lid N02692 96 well adapter for plate w o lid N02693 96 well adapter for plate w lid N02690 384 well adapter for plate w o lid N02691 384 well adapter for plate w lid N06210 1536 well adapter for plate w o lid N02696 6 48 well adapter for plate w o lid N0269
42. position L 76 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Thermo Fisher Scientific Routine Operation Other functions To start the measurement simultaneously to dispensing the dispensing head has to be placed into the corresponding measurement position For example flash luminometric measurements require simultaneous measurement and dispensing However all dispensers and dispensing positions can be used to dispense into any well using any measurement mode s Depending on the dispensing position used for dispensing an extra plate movement may be carried out before the measurement step This may cause minor time delays The instrument automatically recognizes into which dispensing position a dispenser head is inserted The two measurement positions of the instrument are marked F and L Figure 4 50 The readings that are carried out in both of these measurement positions are presented in Table 4 5 Table 4 5 Readings carried out in measurement positions F and L Measurement position F Measurement position L photometric normal and filter mode luminometric fluorometric top fluorometric bottom TRF top TRF bottom spectral luminescence In luminometric measurements two starter reagents are more often used and that is why Dispenser 2 is located on the left hand side and uses dispensing position L2 The dispensers are each equipped with a dispensing head position sensor Figure 4 50 a
43. recommended to service the instrument at least yearly Refer to Service contracts on page 108 If you believe that liquid has entered the Varioskan Flash first switch the instrument off Figure 2 2 and unplug the instrument Carry out corrective measures Refer to How to clean the measurement chamber Thermo Fisher Scientific Maintenance Regular and preventive maintenance on page 89 and Decontamination procedure on page 102 for aid If necessary contact your local Thermo Fisher Scientific representative or the Thermo Fisher Scientific technical service department Refer to How to pack for service on page 107 and Service request protocol on page 129 Although the Varioskan Flash is constructed from high quality materials you must immediately wipe away spilt saline solutions solvents acids or alkaline solutions from outer surfaces to prevent damage Caution If local or laboratory regulations prescribe regular decontamination it is not advisable to use formaldehyde since even small traces of formaldehyde negatively affect the enzyme being used in EIA tests resulting in bad test results A Warning If any surfaces have been contaminated with biohazardous material a mild sterilizing solution should be used A Caution Do not autoclave any part of this instrument A Caution Do not use alkaline or chlorite solutions for cleaning any parts of the measurement chamber Figure 3 25 which may result in
44. section on Do s and Don ts summarizes all the relevant procedures on what to do and what not to do e Incase of any emergencies occurring during operation switch off and unplug the instrument immediately Carry out corrective measures If the corrective measures taken do not help contact authorized technical service e You can push in the tray manually only when the instrument is switched off e Keep all the holes in the adapter clean e Ensure that you select a correct plate type Too high a plate may get jammed and with too low a plate the dispensing might fail and pass by e When placing a microplate onto the tray always make sure that the correct plate type has been selected in SkanIt Software for Varioskan Flash Protocol Properties Plate template before you do anything else e Ensure that the bottom of each microplate is dry Fluid on the bottom of a microplate may present a contamination hazard e Take into account the chemical resistance of the dispensers Table 4 6 and microplates Thermo Scientific Varioskan Flash User Manual 61 Routine Operation Do s and Don ts of the Varioskan Flash Don t 62 Thermo Scientific Varioskan Flash User Manual Determine the compatibility of chemicals used in individual applications with the dispensers Otherwise damage to the dispensers and or test results may occur Ensure that the dispensers are properly installed Otherwise leakage might occur Make sure you do
45. service To execute the alignment sequence a white 384 well plate with 30 pl of luminous solution in well H12 is needed The luminescence intensity of the solution should be in the range of 10 to 108 RLU multiplied with the LumiSens scaling factor For example with the scaling factor 2 7 gives the range of 2 7 x 10 to 2 7 x 10 RLU Any luminescent sample with steady glow type luminescence can be used for the alignment For example the following luminous solutions or samples can be used e Bioluminescent sample with Firefly luciferase ATP reaction e g with Promega s Steady Glo Luciferase Assay System BioThema s Luciferase Assay Kit Promega s ENLITEN ATP kit or BioThema s ATP Biomass kit e Chemiluminescent sample with luminol luminescence e Any other reagent producing a stable luminescence signal Detailed instructions on how the alignment plate can be prepared using Promega s ENLITEN ATP kit are given in Appendix D Preparation of Luminometric Alignment Plate The Varioskan LumiSens measurement position is aligned using the following procedure 1 In SkanIt Software select Settings gt Instrument 2 Select the correct instrument and connect to it You must select the instrument as the default before you can connect to it 3 Click Setup and select the LumiSens module tab 4 Click the Align button 5 The software shows a list of 384 well plates and instructs the user to pipette 30 pl of lu
46. size 1 ml standard 5 ml on request Dispensing tip sizes 0 40 mm standard 0 25 mm on request Dispensing volume 1 ml syringe 1 10 000 pl with 1 pl increments 5 ml syringe 5 10 000 ul with 5 ul increments Automatic safety control based on maximum well volume 1 ml syringe 0 40 mm tip size default 1 ml syringe 0 25 mm tip size Accuracy 5 10 000 pl lt 0 2 ul or 2 whichever is greater Precision 5 19 ul lt 5 20 10 000 ul lt 2 Accuracy 1 20 ul lt 0 2 ul or 2 whichever is greater Precision 1 4 yl lt 10 5 20 ul lt 5 Dispensing speed 96 well plate 30 s 384 well plate 80 s 5 pl well 1 ml syringe 0 40 mm tip Dead volume Tip priming volume lt 100 ul total tubing volume lt 800 ul 1 10 ul Plate sensing Photometric Plate types Safety specifications Safety performance EN 61010 1 2001 Ed 2 6 384 well plates Thermo Fisher Scientific EN 61010 2 010 2003 Ed 2 including US and CA National differences Thermo Scientific Varioskan Flash User Manual 115 Technical Specifications In conformity with the requirements In conformity with the requirements 116 Thermo Scientific Varioskan Flash User Manual The safety specifications are also met under the following environmental conditions in addition to or in excess of those stated in the operating conditions Altitude Up to 20
47. the Varioskan Flash optical measurement modules is shown in the following block diagram Figure 2 6 Each submodule is described separately in the subsequent lower level block diagrams Figure 2 7 through Figure 2 13 Thermo Scientific Varioskan Flash User Manual 23 Functional Description Optical system Principle of the optical system Spectral scanning module LumiSens module Excitation Emission Luminometer optics optics optics Dispensing position F1 Photometer beam through the plate Emission Excitation Fluorometer bottom optics Photometer optics Emission fiber Excitation fiber Figure 2 6 Varioskan Flash optics and measurement positions F and L The Varioskan Flash optical unit consists of six subunits Figure 2 6 e The excitation optics produces light of selected wavelength for fluorometric measurement and also for photometric measurement Refer to Excitation optics on page 25 e The measurement optics produces a high definition optical beam for fluorometric and photometric measurements The excitation light reference detector is incorporated into the measurement optics Refer to Measurement optics on page 26 e The emission optics carries out the reading of a selected wavelength for fluorometry and spectral scanning module based luminometry Refer to Emission reading module on page 27 e The photometric measurement module measures light beam intensity passing through the w
48. the dispensers on page 81 Table 4 6 provides guidelines for compatibility with materials used in the fluid path of the dispensers Compatibility information is based on charts provided by the material manufacturer It is recommended that each laboratory determines compatibility for their respective applications Caution Failure to determine compatibility of chemicals used in individual applications with the dispensers may result in damage to the dispensers and or test results A Plastic materials used in the dispensers Teflon PTFE TFE FEP tubing valve plug and seal Kel F valve body Polypropylene PP fittings for tubing and dispensing tip Silicone tube between dispensing tip and dispensing tube Note Kel F is the brand name for 3M s PCTFE that is polychlorotrifluoroethylene The present brand name is Neoflon CTFE manufactured by Deikon A Caution Take into account the chemical resistance of microplates Most microplates are made of polystyrene that has very limited chemical resistance and should not be used for example with any organic solvents Refer to microplate suppliers documentation regarding the chemical resistance of their microplates A Classification in the table No data available No effect excellent Minor effect good Moderate effect fair Severe effect not recommended oa Ne O Polypropylene satisfactory to 22 C 72 F Polypropylene satisfactory to 49
49. the tray on page 91 SIS sS Change the 8 well plate strip piece i e the tip priming vessel after about 250 tip primings if the priming volume is 10 ul and after 2500 tip primings if the priming volume is 1 ul See Dispensing on page 76 Clean the reagent basin and dispensing area when necessary See v How to clean the reagent basin and dispensing area on page 91 Clean the dispensing base when necessary See How to clean the dispensing base on page 92 Clean the fluorometric bottom reading optics when necessary See How to clean the optics in the measurement chamber on page 90 Clean the photometric optics when necessary See How to clean the optics in the measurement chamber on page 90 SOON NOON Clean the Varioskan LumiSens optics when necessary See How to Continued Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 87 Maintenance Regular and preventive maintenance Item gt n S p pg D g a gt clean the Varioskan LumiSens optics on page 92 Clean the LumiSens base when necessary See How to clean the v Varioskan LumiSens base on page 94 Maintain the dispensers See Routine maintenance of the dispensers v v on page 96 Replace the dispenser input and output tubing when necessary See v v Replacing the aspirate tube assembly or the complete dispensing tube assembly on page 98 Replace th
50. tray is dirty first switch the Varioskan Flash off by pressing the power switch Figure 2 2 on the left side panel of the instrument into the OFF position 4 Then wipe the tray surface and the adjacent instrument surface for example dispensing area Figure 3 34 with a soft cloth or tissue paper moistened with distilled water a mild detergent SDS sodium dodecyl sulfate or soap solution 5 Ifyou have spilt infectious agents on the instrument disinfect with 70 ethanol or some other disinfectant see Decontamination procedure on page 102 6 After cleaning or disinfection push the tray manually into the instrument e Note You can push in the tray manually only when the instrument is I1 switched off amp 84 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Routine Operation Emergency situations Emergency In case there is any abnormal situation during the operation such as r fluids spilling inside the instrument follow these steps situations oe k 1 Switch OFF the instrument Figure 2 2 2 Unplug the instrument immediately from the power supply Figure 3 37 3 Carry out appropriate corrective measures However do not disassemble the instrument 4 If the corrective measures taken do not help contact authorized technical service or your local Thermo Fisher Scientific representative Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 85 Routine Op
51. visit our worldwide Web sites on the Internet at http www thermo com http www thermo com varioskan http www thermo com readingroom In an effort to produce useful and appropriate documentation we appreciate your comments on this user manual to your local Thermo Fisher Scientific representative Thermo Scientific Varioskan Flash User Manual 3 About This User Manual Safety symbols and markings Safety symbols and markings Safety symbols and markings used on the Varioskan Flash Be blelo WARNING UV LIGHT Do not look directly at light gt x 4 Thermo Scientific Varioskan Flash User Manual These symbols are intended to draw your attention to particularly important information and alert you to the presence of hazards as indicated The following symbols and markings appear on the type label and the instrument itself Power ON a Power OFF a Serial number a Catalog number a Date of manufacture a Consult instructions for use a Risk of radiation injury a WEEE symbol This product is required to comply with the European Union s Waste Electrical amp Electronic Equipment WEEE Directive 2002 96 EC A Warning To avoid stray light use blind plugs to close empty dispensing positions A A black label with the following text Figure 2 3 CAUTION WARNING DISCONNECT SUPPLY BEFORE SERVICING and AVERTISSEMENT COUPER L ALIMENTATION AVANT L ENTRETIEN ET LE DEPANNAGE 4 Thermo Fisher S
52. 0 35 40 45 50 Diameter mm Speed and diameter combinations for 24 well plates 1200 1080 RPM 0 5 10 15 20 25 30 35 40 45 50 Diameter mm RPM RPM 1200 1080 960 840 720 600 480 360 240 120 0 1200 1080 960 840 720 600 480 360 240 120 0 Speed and diameter combinations for 12 well plates 0 5 10 15 20 25 30 35 40 45 50 Diameter mm Speed and diameter combinations for 48 to 1536 well plates 0 5 10 15 20 25 30 35 40 45 50 Diameter mm Allowed combination B Recommended combination Figure 4 49 Speed amp diameter combinations for different plate formats 74 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Thermo Fisher Scientific Incubating Routine Operation Other functions Incubation can be carried out with 24 to 1536 well plates The instrument has a universal tray fitted with an adapter that supports incubation Figure 2 19 specially designed for precise and uniform temperature control The incubation allows cellular assays certain enzyme assays and other temperature critical applications to be read under controlled conditions Refer to Incubator on page 31 and Universal tray with adapters on page 35 The incubator consists of two heating elements one below the plate and one above the plate The below the plate element is the universal tray The above the plate element is fixed to the measurement ch
53. 00 m Temperature 5 C to 40 C Humidity Maximum relative humidity 80 for temperatures up to 31 C decreasing linearly to 50 relative humidity at 40 C Mains supply 10 from nominal fluctuations The Varioskan Flash bears the following markings Type 3001 100 240 Vac 50 60 Hz 200 VA CE mark CSA monogram The Varioskan Flash conforms to the following requirements 2006 95 EC Low Voltage Directive 2004 108 EC Electromagnetic Compatibility Directive EMC FCC Part 15 Subpart B Class B July 2004 2002 96 EC Waste of Electrical and Electronic Equipment EMC performance EN 61000 6 3 2001 Generic standards Emission standard for residential commercial and light industrial environments EN 61000 6 1 2001 Generic standards Immunity standard for residential commercial and light industrial environments EN 61326 1 1997 A1 1998 Product family standard A2 2001 A3 2003 Thermo Fisher Scientific Thermo Fisher Scientific Test standards EN 55022 1998 A1 2000 Technical Specifications In conformity with the requirements Performance limits Class B 150 kHz 1 GHz EN 55014 1 2000 EN 61000 3 2 2000 EN 61000 3 3 1995 A1 2001 ANSI C63 4 2000 Class B 0 150 0 550 1 4 30 MHz Class A Class B 450 kHz 1 GHz 2002 EN 61000 4 2 1995 A1 1998 A2 2001 4 kV CD 8 kV AD Criteria B EN 61000 4 3 1996 A1 1998 3 V m 80 MHz 1 GH
54. 05 Only numbers are entered for example 0614 05 Thermo Scientific Varioskan Flash User Manual 55 Installation Installation setups Varioskan LumiSens measurement position alignment 56 Thermo Scientific Varioskan Flash User Manual 5 The calibration is started and will take approximately a minute to complete 6 It is also possible to run the LumiSens calibration manually with SkanIt Software The need for manual calibration can be decided based on the following rules a Calibration should be run if the LumiSens module has been used with Varioskan Flash before but another unit than the previously used one is installed into the Varioskan Flash b Calibration should be run if the same LumiSens module is installed after service or when any mechanical or electronic replacements have been done to the LumiSens module c Calibration is not necessary if the LumiSens module has been removed for normal routine maintenance for example cleaning and so on The Varioskan LumiSens calibration is started using the SkanIt Software menus Settings gt Instrument gt Setup and selecting the Varioskan LumiSens tab From the LumiSens dialog select Calibrate The software will display the existing serial number of the LumiSens module type the correct serial number or accept an existing one and start the calibration with OK 7 When the calibration is completed it is recommended to continue the installation process with the Va
55. 17 41 64 Thermo Fisher Scientific check 17 41 64 optical 23 24 25 27 62 64 87 90 93 136 145 146 density 136 145 146 measurement modules 23 system 23 24 62 87 orbital shaking 32 73 111 115 order sorting 146 ordering information 3 35 38 39 131 P packing 41 42 107 108 129 instructions 42 107 list 42 materials 108 pathlength correction 120 121 122 calculation 121 factor 121 photometric measurement 15 23 24 26 28 33 70 71 module 24 28 photometric spectrum scanning 71 photometry 16 25 66 111 113 119 146 photomultiplier tube 27 29 114 127 146 plate 15 33 34 35 36 37 65 66 111 112 113 114 115 119 120 131 adapter 32 33 34 35 36 37 39 48 49 61 65 75 91 120 131 145 sensing 115 template 61 65 126 tray 33 34 35 37 38 39 48 49 91 120 127 145 type 35 49 61 65 112 113 114 115 121 128 plunger 100 127 holder arm 100 lock screw 100 position calibration hole s 32 35 positioning lever 34 48 49 66 precision 97 113 115 146 prime 31 34 38 50 62 63 78 79 80 84 87 96 97 102 107 126 127 146 Prime Empty switch 31 78 80 priming 31 34 38 50 62 63 78 79 80 84 87 102 107 126 146 vessel 34 38 63 78 79 84 102 107 volume 62 78 79 87 protocol 61 65 66 67 pull back feature 79 Thermo Scientific Varioskan Flash User Manual 151 Q quantificati
56. 2 4 Close up of the computer and mains supply connectors 20 Figure 2 5 Varioskan Flash internal views 20 Figure 2 6 Varioskan Flash optics and measurement positions F and L 24 Figure 2 7 Excitation Optics 20 0 ccccccscecessescsescetesesesteteseseeteteseseseeesees 25 Figure 2 8 Principle of the double monochromatOl ccccceccesee eects 26 Figure 2 9 Measurement optics oo ccc cece cscetesesescecsteteseseetetesesesteenees 27 Figure 2 10 EMISSION OPtiCS 00 0 0 cccce cc cececesesesceteseseecetesesesteesesesesesteneeees 27 Figure 2 11 Photometric measurement module uu cece cece teesteeete teens 28 Figure 2 12 Fluorometric bottom reading optics 20 0 cece ceteeseeteees 28 Figure 2 13 Varioskan LumiSens measurement MOdUIE 0 cccceee 29 Figure 2 14 Varioskan Flash dispensing system ccccccceceesceteseseseeteees 30 Figure 2 15 Dispensing tip options 0 40 mm and 0 25 MM uuu cece 31 Figure 2 16 Control switches 0 cccceccccccccceseseesssestesestesestesesteseeteseeteeeees 31 Figure 2 17 Varioskan Flash incubator cross section ccccccecesesceteees 32 Figure 2 18 Part of the Varioskan Flash track mechanisM ccccce 33 Figure 2 19 Assembly picture of the universal tray 05110010111011151115110 34 Figure 2 20 Robotic tray fitted with adapter for plate w o lid 126 38 Figure 2 21 Changing the location of the tip priming vessel holder
57. 36 well plates using the universal tray and a correct adapter You need different adapters for plates with or without lids The universal tray with a 6 to 48 well plate adapter supports measurement but not incubation An adapter that supports incubation for 24 to 48 well plate formats can be ordered on request Note The efficiency of incubation may be remarkably reduced with large well diameter plates A Caution Do not use plates with dimensions exceeding the top rim of the tray Note that the maximum total height of plates is manufacturer related A Refer to Table 2 1 and Chapter 9 Ordering Information For more information on plate type settings refer to the SkanIt Software for Varioskan Flash User Manual Cat no N06355 Thermo Scientific Varioskan Flash User Manual 35 Functional Description Tray options ID no 2 Picture Table 2 1 Compatibility of the universal tray and plate specific adapters Description Measurement Incubation Dispensing 96 well adapter for plate without lid yes yes yes To be used with the most common 96 well plate formats without lids Incubation and dispensing are possible 3 96 well adapter for plate with lid yes yes no To be used with the most common 96 well plate formats with lids Incubation is possible No dispensing 4 384 well adapter for plate without lid yes yes To be used with the most common 384 well plate formats without lids Incubation and dis
58. 43 How to release the transport locks ssssseeeesseeesesesererrrererersrerssss 43 Tray holder transport lock s ssssssessessesssssessssssrsrsesrsreererersrerseees 44 Varioskan LumiSens optics transport lock sssssessseeeseseeeseseessss 47 How to install the tray sssessesessesesssesseseseseesesesseseseseesesesssseseseeeesese 48 How to setup the dispensers ssssssssssssssesreesesrsrsrsrsererrrererereressses 49 Installation of SkanIt Software for Varioskan Flash eee 51 Cable Cori me tenn sist Seales cg etesen A EE 51 Varioskan LumiSens module installation ceeeeeeeeeereeeeeenees 52 Varioskan LumiSens measurement position alignment 56 Varioskan LumiSens scaling factor 2206 26 4 desea 58 Varioskan LumiSens module filter installation eee 58 Routine Operation sssssssssssssnnnessseusunnnnnnssuununnnnnnounnnnnenounnunnnnneonnnnnnnnnenennnnn 61 Do s and Don ts of the Varioskan Flash ssseeeseseseseeereeererrrrerereeree 61 DE A E odes tas Roos E tines ttnantg Uaesuu cts Bucs ES 61 Don ternpi n a E A T E ARA A 62 SWItehINE ON eeose okena at eSEE aane e asear 63 Operationaleheckiinosnennan sann en n R RR a R 64 Loading the iiicroplate sacs cigucan shasta Mahe a A oa esia 65 Automatic runtime calibrations sacs ccs eatin oneev ea eteae 66 Fluorometric measurements sis cseeien dei detente ceed 67 Fluorometric spectrum stan nine caccesiedccctee ci cakes cue cetene ae cackiae 70 Photometric
59. 6 well or a 384 well plate Q9 Can I enter my own pathlength correction factor for a reagent not found in the preset list A9 Yes this can be done in SkanIt Software for Varioskan Flash from the Edit Pathlength Correction K Factors dialog Settings gt Pathlength Correction Q10 Why would I need to use a pathlength correction factor A10 Pathlength correction is required to be able to calculate concentrations using Bouguer Lambert Beer s law with molar absorption coefficients from the literature Q11 How do I generate my own pathlength correction factor A11 To generate a new pathlength correction factor refer to the SkanIt Software for Varioskan Flash User Manual Cat no N06355 Note The pathlength correction is dependent on the material used to manufacture the plate and should be redetermined if the plate type is changed A Q12 Can plates be used to directly measure the concentration of DNA or proteins A12 Yes with plates suitable for UV measurements down to 260 nm Q13 How long does it take for the Varioskan Flash incubator to reach 37 C A13 The incubator reaches 37 C in approximately 5 minutes from 25 C However warming up solutions in the microplate takes about 1 hour Q14 What is the useful life of the lamp A14 Typically 10 flashes of reading of 1 million microplates 96 wells by using the 100 ms measurement time Thermo Scientific Varioskan Flash User Manual 121 Frequently
60. 62 88 94 95 107 128 module 15 24 28 29 41 43 52 55 58 59 60 71 95 127 128 module filter installation 58 module installation 52 55 Thermo Fisher Scientific optics 47 53 55 87 92 93 94 104 106 107 114 optics transport lock 47 104 106 W warm up time 114 warning codes 64 125 128 warranty 2 42 62 129 Warranty Certificate 42 wavelength 16 21 22 24 25 26 66 70 71 112 113 114 119 122 128 145 146 accuracy 112 113 114 range 16 113 114 119 selection 25 112 113 114 Web links 133 134 Thermo Scientific Varioskan Flash User Manual 153 154 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Notes Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 155 Notes 156 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Notes Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 157 Thermo Fisher Scientific Oy Microplate Instrumentation Ratastie 2 P O Box 100 Fl 01621 Vantaa Finland www thermo com N06354
61. 7 6 48 well adapter for plate w lid N03395 96 well adapter for PCR plate w o lid N03079 Robotic tray incl adapter for plate w o lid N02339 Adapter for plate w o lid SP 00094 Dispensing tube assembly 0 40 mm for Varioskan Flash N03080 Dispensing tip 5 1000 ul 0 40 mm 10 pieces N03081 Dispensing tip 1 20 pl 0 25 mm 10 pieces Continued Thermo Scientific Varioskan Flash User Manual 131 Ordering Information Upgrade kits Cont Code Item Quantity 95029350 1x8 Microtiter Solid Strip Assembly 50 pieces Alternatively the following codes can also be used 95029100 95029450 95029490 and 95029510 2805690 Aspirate tube assembly incl tubing and end weight SP 00096 Dispenser syringe 1 ml SP 00093 Dispenser syringe 5 ml SP 00100 3 port valve 1 4 28 fittings 24071700 Bottle stand N06494 10 00 PQ Varioskan Flash Document N03394 Spectrophotometric verification plate 2806460 Lumiwell verification plate 2305290 RS 232C interface cable D9 Female D25 Female N04001 USB A B device cable other plate specific adapters for universal tray available on request longer USB cables available from PC stores Upgrade kits Table 9 22 Codes for upgrade kits Code Item Quantity 2806350 Dispenser upgrade kit for Varioskan Flash w 1 ml syringe 1 0 40 mm tip N06492 Varioskan LumiSens upgrade kit 1 132 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientifi
62. 9 70 112 113 121 122 126 128 LED indicator 6 63 64 146 lever opening bar 48 light source 25 28 64 69 70 112 113 121 122 126 128 literature 135 lock knob 53 55 93 94 95 Low range 67 68 72 Thermo Fisher Scientific luciferase 23 57 146 luminescence 15 16 21 22 28 57 71 77 119 120 134 135 136 145 146 luminometric 15 23 33 67 71 72 73 77 79 146 label 146 measurement 15 23 33 71 73 77 79 spectrum scanning 73 luminometry 16 24 67 111 114 luminophore 146 LumiSens 52 55 56 57 58 60 88 alignment 56 57 58 95 143 M mains 20 41 51 52 103 111 116 power supply connector 52 switch 51 maintenance 3 6 62 76 80 87 88 95 96 97 106 108 129 checklist 62 87 means of identification of tray adapter combinations 34 measurement 67 68 69 70 71 72 73 chamber 6 32 44 47 62 63 70 71 74 75 77 87 89 90 91 96 104 105 127 chamber door 6 44 47 62 63 70 71 89 90 104 105 127 optics 24 26 27 position 24 26 28 30 56 57 58 76 77 95 speed 25 112 113 114 techniques 21 112 135 146 time 68 69 70 72 112 113 114 121 type 111 Medium 27 67 68 72 high range 67 68 low range 67 range 72 molar absorption coefficient 22 121 122 monochromators 25 26 27 71 72 146 multiplexing 146 NIR 15 16 nonvolatile memory 64 67 71 0 ON OFF switch 100 operational
63. A Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 93 Maintenance Regular and preventive maintenance How to clean the Varioskan LumiSens base 94 Thermo Scientific Varioskan Flash User Manual Figure 5 55 Cleaning the Varioskan LumiSens optics 6 Replace the Varioskan LumiSens optics unit Figure 3 42 7 Refasten the green lock knob To clean the Varioskan LumiSens base follow these steps 1 Switch off the Varioskan Flash by pressing the power switch Figure 2 2 on the left side panel of the instrument into the OFF position 2 Place the dispensing heads into the dispensing head holders 3 Remove the Varioskan LumiSens optics Figure 5 54 by unfastening the green lock knob Turn the lever and lift the optics off 4 Clean the Varioskan LumiSens base from the top by soaking a cotton stick in a mild detergent solution soap solution or 70 ethanol Figure 5 56 Do not use formaldehyde Thermo Fisher Scientific Maintenance Regular and preventive maintenance Figure 5 56 Cleaning the Varioskan LumiSens base from above 5 To expose the crosstalk shield further unfasten the two fastening screws and turn the part over Do not unfasten the electronic connections 6 Carefully clean the dirt from the bottom using a lintfree tissue or a cotton stick soaked in a mild detergent solution soap solution or 70 ethanol Figure 5 57 Do not use formaldehyde
64. C 120 F Thermo Scientific Varioskan Flash User Manual 81 Routine Operation Other functions 82 Thermo Scientific Varioskan Flash User Manual Table 4 6 Compatibility chart of solvents suitable with the plastic materials used in the dispensers Solvent Acetaldehyde Teflon 0 Kel F Polypropylene 0 Acetates 0 Acetic acid 0 Acetic anhydride Acetone 0 0 0 0 0 Acetyl bromide Ammonia Ammonium acetate Ammonium hydroxide Ammonium phosphate Ammonium sulfate Amyl acetate Aniline Benzene Benzyl alcohol Boric acid Bromine Butyl alcohol Butyl acetate Carbon sulfide Carbon tetrachloride Chloroacetic acid Chlorine OoO oO oT oy Toy Toy Toyoyoyoyoylyo Chlorobenzene Chloroform Chromic acid Cresol Cyclohexane Dimethyl sulfoxide DMSO Ethers Ethyl acetate Ethyl alcohol Ethyl chromide Formaldehyde Formic acid oO To To Toy Toy oyosypoywoyoy y mo Continued Thermo Fisher Scientific Routine Operation Other functions Cont Solvent Teflon KelF Polypropylene 0 Freon 0 Gasoline Glycerin Hydrochloric acid OO a Hydrochloric acid conc Hydrofluoric acid Hydrogen peroxide Hydrogen peroxide conc Hydrogen sulfide OoO OoO or E N EE e E E a E a Kerosene Methyl ethyl ke
65. ON the color indicates the state of the instrument Table 4 4 Table 4 4 Front panel indicator light LED Instrument status Green The instrument is ready and waiting for a command from SkanIt Software for Varioskan Flash Orange O The instrument is busy executing startup functions or commands from Skanlt Software for Varioskan Flash Red C All error messages are stored in the internal memory log file of Varioskan Flash The error log file can be accessed with Skanlt Software by selecting Settings gt Instrument gt Setup gt Reports gt Error Log gt Run Report Warning Never operate your instrument from a power outlet that has no ground connection A Switch the Varioskan Flash on by pressing the power switch on the left side panel of the instrument into the ON position Figure 2 2 At startup the instrument is automatically checked for correct operation Startup routines are performed Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 63 Routine Operation Switching on Operational check First switch the Varioskan Flash ON Figure 2 2 The instrument has a sophisticated control system The instrument automatically performs a complete set of initialization tests and adjustments The mechanical electrical and optical functions of the instrument are checked at startup for example e instrument configuration e tray positioning e measurement selectors positioning e exci
66. RET assays aspirate dispense tubing Connects the valve output port 1 4 28 thread to a sample source and destination The aspirate tubing is used to fill the syringe with reagent The dispensing tube is used to dispense reagent from the syringe into a microplate bioluminescence Naturally occurring chemiluminescence from light emitting organisms e g glowworms some deep sea fish some bacteria and some fungi BRET Bioluminescence resonance energy transfer Refer to Luminescence on page 22 chemiluminescence Luminescence as a result of pure chemical reactions decade Order of magnitude A logarithmic value that is used for presentation of dynamic range decontamination Removal or neutralization of radiologic bacteriological chemical or other contamination DELFIA Dissociation enhanced lanthanide fluoroimmunoassay PerkinElmer s product family for TRF reagents and kits diffraction Spreading of light transmitted through a narrow slit or reflected from a narrow groove diffraction angle An angle between incoming and diffracted light which is the function of the wavelength and groove spacing diffraction grating reflection grating Grooved component that diffracts the incoming light into diffraction angles Thermo Fisher Scientific disinfection The destruction of pathogenic bacteria usually with an antiseptic chemical or disinfectant double monochromator Two serially one after another connected
67. The normal mode uses the Varioskan LumiSens module without placing any filters in the lightway The filter mode uses user defined and installed filter s The monochromator uses the spectral module emission optics to select the correct wavelength s Note that this mode is considerably weaker in sensitivity performance 6 The dynamic range Figure 4 46 is selected by the user e AutoRange e Manual range for the luminometric normal and filter optics is Thermo Scientific Varioskan Flash User Manual 71 Routine Operation Luminometric measurement High range Medium range e Low range e Manual range for the luminometric monochromator optics is equivalent to the fluorometric measurement Refer to Figure 4 46 a Automatic dynamic range selection AutoRange default selects automatically that the optimal reading range is used It is based on signal intensity in the well and uses the lowest possible reading range to obtain best sensitivity However if you need the fastest possible operating speed use the manual dynamic range selection of Low Medium or High range b Select manual dynamic range according to the following principles High range is intended for highest concentration samples It covers a wide dynamic range with somewhat lower sensitivity than with other dynamic ranges Low range produces on the other hand the highest sensitivity with a limited dynamic range When selecting a fixed gain the princip
68. amber ceiling Both heating elements are temperature controlled Note The samples in the microplate reach the target temperature usually much later than the instrument amp Thermo Scientific Varioskan Flash User Manual 75 Routine Operation Other functions Dispensing This section provides valuable information on how to use and maintain the optional dispensers Caution Make sure you do not dispense into the instrument by mistake Ensure that e A correct microplate has been inserted e The microplate or tip priming vessel is not too full e The recommended Check plate in SkanIt Software for Varioskan Flash Settings gt Options is ticked A The Varioskan Flash can be equipped with three optional dispensers Figure 2 5 allowing dispensing from 1 to 10 000 ul in 1 pl increments with the 1 ml syringe and dispensing from 5 to 50 000 ul in 5 pl increments with the 5 ml syringe Dispenser 2 Dispenser 3 Dispenser 1 9 3 OO Measurement position F Measurement H position L ee Dispensing Dispensing position L2 position L1 Figure 4 50 Identification of the dispensing tip position The three dispensing positions are marked F1 L1 and L2 Figure 4 50 The F1 and L1 dispensing positions can be used with either Dispenser 1 or 3 Dispensing position F1 points the dispensing head towards the measurement position F Dispensing positions L1 amp L2 point the dispensing heads towards the measurement
69. amber door opened 2 Lift up the front cover of the instrument from both sides and remove it to make the transport lock accessible Figure 3 25 44 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Thermo Fisher Scientific Installation Installation setups Lift the front cover away from both sides Figure 3 25 Front cover removed You will notice that the metallic transport lock support is fastened with four screws marked 1 2 3 and 4 Figure 3 26 The two top screws marked 1 and 2 are fastened to the tray holder The two bottom screws marked 3 and 4 are fastened to the track mechanism bar 3 Unscrew counterclockwise the two top screws marked 1 and 2 of the transport lock with the hexagonal screwdriver supplied Figure 3 26 so that the track mechanism is loosened Remove screws 1 and 2 and store the transport lock tag Figure 3 26 Transport lock fastened screws 1 4 shown 4 At the same time remove the transport lock tag from the topmost screw marked 1 Figure 3 26 Keep the tag for future relocation or transportation of the instrument 5 Gently push the track mechanism into the instrument by hand Thermo Scientific Varioskan Flash User Manual 45 Installation Installation setups 46 Thermo Scientific Varioskan Flash User Manual 6 Fasten the two top screws marked 1 and 2 clockwise to the two tapped holes located on the track mechanism bottom plate
70. asurement module cccccccceeseeeeeees 28 Disp nseis Sienan a AN ee AU tae agate tod een ian 29 Control switehes tannanna r E E A ONE TS 31 GUD AHO A E T EEA E EE 31 Track mechanism sseessseeseeessessereeereessresstessreesrresreesseessreesrresereeses 32 Trayoptiotscresnnpnnnu aiena e a aaia 33 Eray COMPOS IION a sos cerechdedese sedersnsoewandeleca wvenigensaecusvoivesiiales 34 Universal tray with adapters aooSietacsandeb cS sceded contac vondtetcteamneataes 35 Robotic tray seeren a E E R A R 37 How to change the location of the holder for the tip priming AN ARRE E T A EE E I E E E AA 38 How to remove or replace the robotic tray adapter 39 Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 7 Contents Chapter 3 Chapter 4 8 Thermo Scientific Varioskan Flash User Manual nstallati oi adsapdnedadsdusttespauundeseeasnccansonstuontnanard y Ipstallation checkliste nsh oni nn i EN RR 4 What to do upon delivery siete conc vendeitnczaendvclcnelel eats behav eiaiteans 42 How to unpack seserinis 42 Checking delivery for completeness ya cscvvessentnstnondveousesuedvactedy es 42 Checking for damage during transport cseseceeeeesseeseeeeeeseees 42 Environmental requirements et voce lat ao heat Oa tas 43 TNIV HAV Oe 2 62a rgd tater lea an at ai 2 een Maries ei 43 Technical prerequisites csccsaccsie ss deettinazasetiteesientaccsiaraicveiduatenieaee 43 Installation Set PS ioiii rishun inei rinas
71. ated automatically A typical calibration time when less than 10 wavelengths are used is a few seconds but the calibration for a spectrum scan from 200 to 1000 nm with a 1 nm increment takes over 1 5 minutes An automatic runtime calibration expire period is one hour in photometry Recalibration is performed 45 minutes after the previous calibration depending on the measurement procedure If any measurement is performed later than one hour after the previous calibration a warning appears about calibration validity This may happen if an assay takes a long time to execute and there is no sufficient time slot to perform calibration during protocol execution However the warning does not mean that the measurement failed only that the accuracy of the results may have suffered In fluorometry the automatic calibration expire period is 10 minutes Recalibration is performed 7 5 minutes after the previous calibration depending on the measurement procedure If a measurement is performed 10 minutes after the previous calibration a warning appears about calibration validity This may happen if an assay takes a long time to execute and there is no sufficient time slot to perform calibration during protocol execution However the warning does not mean that Thermo Fisher Scientific Routine Operation Fluorometric measurement the measurement failed only that the accuracy of the results may have suffered In luminometry the automatic calibration
72. ation procedure under the topmost screw marked number 1 before tightening the screw marked number 2 Figure 3 26 8 Tighten the bottom screws marked number 3 and 4 firmly once you have fastened the top screws marked number 1 and 2 of the transport lock Figure 5 63 B 9 Replace the front cover Figure 2 2 and Figure 5 51 and close the dispenser sliding cover and measurement chamber door Figure 3 23 Varioskan LumiSens To refit the Varioskan LumiSens optics transport lock follow these optics transport lock steps 1 Unfasten the three locking screws from their places of storage on the instrument and turn the gray transport lock counterclockwise 90 Figure 5 64 Locking screw removed and stored Figure 5 64 Varioskan LumiSens optics transport lock unfastened 2 Fasten the two lower screws and the top screw of the Varioskan LumiSens optics transport lock Figure 5 65 Locking screw fastened J ka Figure 5 65 Varioskan LumiSens optics transport lock fastened 106 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Maintaining a system log How to pack for Thermo Fisher Scientific service A Maintenance Maintaining a system log A system log which includes a short summary of the use maintenance procedures error messages and other information about the use of the system can be very useful in properly maintaining the system The information in the log
73. b in the SkanIt Software instrument setup Settings gt Instrument gt Setup Refer to the SkanIt Software for Varioskan Flash User Manual Cat no N06355 The filter installation procedure is carried out using SkanIt Software for Varioskan Flash Refer to the SkanIt Software for Varioskan Flash User Manual Cat no N06355 To carry out filter installation follow these steps 1 Connect SkanIt Software to the Varioskan Flash instrument with the Varioskan LumiSens module installed 2 In SkanIt Software use the SkanIt menus Settings gt Filters to open the Filter Setup dialog Select the LumiSens tab 3 Use the arrows or select a row in the table to select the filter position to be used This will make the selected filter position in the filter wheel to rotate so that the position is placed under the filter installation hole on the Varioskan LumiSens module 4 Press the Change button on the LumiSens tab Thermo Fisher Scientific Installation Installation setups 5 The Change Filter dialog is opened with the filters in the filter library listed Select a filter and press OK You can define new filters by pressing the Add New Filter button 6 To insert the filter into the instrument open the green lid on the Varioskan LumiSens module The selected filter position should be completely visible 7 Pull up the green filter lift at the front of the Varioskan LumiSens module Figure 3 43 BA Figure 3
74. ble 4 6 Compatibility chart of solvents suitable with the plastic materials used in the CISPENSETS c cc cecesesesceteteseeteteeseeteees 82 Table 5 7 Maintenance checklist c cccceccceeecseseeseseseesseeseseesssesseeeeseees 87 Table 5 8 Example of a system 0g ss ssissisnissisrisrrsrrenirenienrenn 107 Table 6 9 General specifications 0 100110011sssirssiiieririssiriesirreerreninen 111 Table 6 10 Fluorometry 0 cece ce cecsescetesesestetesesesteteseseeteteseseeeees 112 Table 6 11 Photometry 2 ccc ceceesescecesesestetesesestetesesestetessseeeeees 113 Table 6 12 Luminometry a sue cesses cutee dace ta cede near viegsadi cede sacs 114 Table 6 13 Incubator so cess b cs ssctess castecds trav a beate dacterndestasketrgeseee beakes 114 Table 6 14 Shaker e ccttaicitiictaceindarndetanieniademeateietidennee 115 Table 6 15 Dispensers sc cceciec ce cteca ccarsacnsesceesea mes ledde Srsnsce soncedeaucnacareeten 115 Table 7 16 Plates used with the Varioskan Flash cc ceceeeeeesteeeeeees 120 Table 7 17 Tray vs plate specific adapter ccc ce ceseeceteteseeeeees 120 Table 8 18 Error codes reported oo ccc cececscecesesesesesteteseseeteteseseseeees 125 Table 8 19 Warning codes reported cececcscecesesestetetesesteteeseeeeees 128 Table 9 20 Instrument catalog NUMDETS 0 0 cece ce teteesteteteeeeeees 131 Table 9 21 Codes for spare parts and accessories 5111051110511105111201 131
75. bstances subject to the RoHS Directive are available at www thermo com WEEERoHS A Regarding the original packaging and packing materials use the recycling operators known to you For more information contact your local Thermo Fisher Scientific representative Thermo Scientific Varioskan Flash User Manual 109 Maintenance Disposal of the instrument 110 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific General specifications Chapter 6 Technical Specifications Thermo Fisher Scientific reserves the right to change any specifications without prior notice as part of our continuous product development program Table 6 9 General specifications General specifications Overall dimensions ca 540 mm W x 580 mm D x 500 mm H 21 3 W x 22 8 D x 19 7 H Weight 55 67 kg 121 148 Ibs depending on the configuration Operating conditions 10 C to 40 C maximum relative humidity 80 for temperatures up to 31 C decreasing linearly to 50 relative humidity at 40 C Indoor use only Transportation conditions Storage conditions Mains power supply 40 C to 70 C packed in transport packaging 25 C to 50 C packed in transport packaging 100 240 Vac 50 60 Hz nominal Power consumption 200 VA max Heat dissipation 683 BTU max User interface The instrument is under PC control and run on Skanlt Software for Varioskan Flash whic
76. c Useful web links Thermo Fisher Scientific Chapter 10 References Additional information on chemical resistance of labware can be found for example from the following Web sites http www thermo com eThermo CMA PDFs Articles articlesFile 15205 pdf see p 63 http www wheatonsci com http www resistoflex com diachem htm http www copely com products acrobat chemical pdf http www nalgenunc com http www tangram co uk TI Polymer Introduction html For more information on microplate plastics manufacturers visit the following Web sites Thermo Fisher http www thermo com Scientific Bibby Sterilin http www bibby sterilin co uk internet BibbySite nsf framesette r TMAY5BQDST OpenDocument Porvair Sciences http www porvair sciences com Corning Costar http www corning com lifesciences Greater_Euro pe en BD Falcon http www bdbiosciences com index_us shtml Nunc http www nuncbrand com Greiner Bio One http www gbo com For more information on reagents and kits visit the following Web sites BD Clontech http www clontech com clontech BD GenTest http www gentest com Thermo Scientific Varioskan Flash User Manual 133 References Useful web links 134 Thermo Scientific Varioskan Flash User Manual DiscoverX http www discoverx com Molecular http probes invitrogen com Probes Roche Applied http www roche applied science com index jsp Science
77. careful to remount it into the original position without using extensive force Do not wash any electronic parts A Caution Try to avoid loosening the crosstalk shield but if necessary be i Figure 5 57 Cleaning the Varioskan LumiSens base from the bottom Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 95 Maintenance Regular and preventive maintenance Routine maintenance of the dispensers Daily maintenance Weekly maintenance 96 Thermo Scientific Varioskan Flash User Manual 7 Replace the bottom part of the Varioskan LumiSens base with the two fastening screws 8 Replace the Varioskan LumiSens module with the green lock knob 9 Run the Varioskan LumiSens measurement alignment Refer to Varioskan LumiSens measurement position alignment on page 56 To obtain optimal performance and maximum useful life for the dispensers Figure 3 34 it is important that the recommended cleaning maintenance instructions are followed Refer also to Dispenser washing on page 80 For use of the Wash functionality refer to Washing dispensers in Chapter 5 of the SkanIt Software for Varioskan Flash User Manual Cat no N06355 The Varioskan Flash is a very sensitive instrument Therefore take special care to avoid any contamination of any parts of the dispenser tubing and follow all GLP Good Laboratory Practices recommendations The basic maintenance procedure should be perf
78. cientific About This User Manual Safety symbols and markings Warning and other The following symbols and markings appear in this user manual markings used in the documentation Warning Risk of electric shock A Warning Biohazard risk A Warning Hot surface risk of burns A Warning Risk of injury to the user s A Warning Risk of ultraviolet radiation injury A Caution Risk of damage to the instrument other equipment or loss of performance or function in a specific application A Note Marks a hint important information that is useful in the optimum operation of the system or an item of interest A H DPS PPe gt Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 5 About This User Manual Instrument safety and guidelines for use Instrument safety and guidelines for use 6 Thermo Scientific Varioskan Flash User Manual Always follow basic safety precautions when using the Varioskan Flash to reduce the risk of injury biohazardous contamination fire or electrical shock Read this user manual in its entirety prior to operating the instrument Failure to read understand or follow the instructions in the manual may result in damage to the instrument injury to laboratory and operating personnel or poor instrument performance Observe all Warning Caution and Note statements as well as safety symbols and markings on the instrument and in the documentation Never open any ot
79. d with 70 ethanol 5 Place the instrument in a large plastic bag Ensure that the dispenser sliding cover and front cover Figure 3 25 are open and the tray is out Figure 4 45 6 Place a cloth soaked in the prepared solution into the bag Ensure that the cloth does not come into contact with the instrument 7 Close the bag firmly and leave the instrument in the bag for at least 24 hours Thermo Scientific Varioskan Flash User Manual 103 Maintenance Decontamination procedure 8 Remove the instrument from the bag 9 Clean the instrument using a mild detergent 10 Remove any stains using 70 ethanol 11 After performing this decontamination procedure enclose a signed and dated Certificate of Decontamination both inside the transport package and attached to the outside of the package How to refit the When you relocate the instrument or ship it for service make sure you transport locks refit the two transport locks the transport lock of the tray holder and the Varioskan LumiSens optics transport lock Tray holder transport Note that the tray holder transport lock support piece is easily lock recognizable having a metallic color and a yellow label Figure 3 26 Refer to How to release the transport lock on page 43 arrows in the opposite directions than in the figures 1 Unfasten the attached tray from the screw fix of the tray holder Figure 3 32 2 Push the tray holder gently into the instrument b
80. d from cell lysates for example photometric fluorometric beta galactosidase alkaline phosphatase and CAT assays Q18 How ofien do I have to run the Varioskan Flash spectrophotometric verification plate A18 It is not necessary to read the Varioskan Flash spectrophotometric verification plate to ensure that the instrument is functioning properly Startup checks for correct instrument operation are performed each time it is turned on However the verification plate is an external second source that documents instrument functionality The frequency with which the verification plate is used depends on the standard operating procedures of each individual user 122 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Frequently Asked Questions Q amp As Q19 What is the minimum time between measurements in kinetic measurements A19 10 ms in one well Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 123 Frequently Asked Questions Q amp As 124 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific warning codes Code 1 Chapter 8 Troubleshooting Guide Note Do not use the instrument if it appears that it does not function properly A Note that the instrument does not verify the logic flow of the received commands Error and When an error is detected the current operation is terminated After an are presented below Table 8 18 Error codes repor
81. dispense when the dispenser is not primed Prime the dispenser before dispensing 67 The dispenser is not installed Do not try to use a nonexistent dispenser Contact service if the dispenser exists 68 The drift compensation factor is too far from the nominal value of Contact service 1 0 69 Not enough memory for a new user defined parameter Delete some unused user parameters to make room for new parameters 75 Default PMT voltages calibration failed Contact service 76 Automatic dynamic range PMT voltages calibration failed Contact service 77 The dark level signal on some of the A D converter board input Contact service channels is too high 78 PMT linearity calibration failed Contact service 79 Bandwidth factors calibration failed Contact service 80 The measurement chamber door is open See that nothing is preventing the measurement chamber door from closing when the plate is driven in Do not open the door while the instrument is measuring 81 Such an error during startup that the execution of some functions The measurement chamber door must be is not possible closed during startup If it is not contact service 82 Invalid TRF measurement command Contact service Continued Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 127 Troubleshooting Guide Error and warning codes Cont Code Explanation Suggested action 83 Varioskan LumiSens crosstalk shield position is incorrect
82. e Empty switch is only present if there is a dispenser fitted For the Prime function a microplate must be loaded and this is automatically checked before priming The Empty function does not require a microplate to be present The Prime Empty switch has an additional function as a washing option of the dispenser Refer to Dispenser washing on page 80 PLATE IN OUT control switch DISPENSER PRIME EMPTY control switch Figure 2 16 Control switches The accurate incubator is useful for temperature critical applications for example certain enzyme assays and cell based applications The universal tray is specially designed for precise and uniform temperature control The incubator consists of two main parts a fixed Thermo Scientific Varioskan Flash User Manual 31 Functional Description Track mechanism Upper heater slightly tilted Top reading window Track mechanism 32 Thermo Scientific Varioskan Flash User Manual upper heater Figure 2 17 and a moving universal tray Figure 2 17 The microplate Figure 2 17 is surrounded by temperature controlled heaters with the exception of a narrow space between the upper heater and the universal tray and the reading windows Figure 2 17 The tray and the upper heater together form the isothermal chamber Note that incubation can be carried out with the Varioskan Flash universal tray and a correct adapter Refer to Universal tray with adapters on page 35 The uppe
83. e Medium high range e Medium low range e Low range The measured values are comparable regardless of the dynamic range selection AutoRange or any of the fixed manual ranges Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 67 Routine Operation Fluorometric measurement 68 Thermo Scientific Varioskan Flash User Manual High range Medium high range Medium low range 0 01 0 1 1 10 100 1000 10000 Concentration decades Figure 4 46 Dynamic range selection a Automatic dynamic range selection AutoRange default selects automatically the optimal reading range It is based on signal intensity in the well and uses the lowest possible reading range to obtain best sensitivity However if you need the fastest possible operating speed use the manual dynamic range selection of Low Medium low Medium high or High range Overrange values can sometimes also occur when the settle delay is too small To allow the liquid surface to settle down for example with a 96 well plate a settle delay of 200 to 400 ms can be necessary depending on the liquid used For more details refer to Settle delay on page 73 a Select manual dynamic range according to the following principles High range is intended for highest concentration samples It covers a wide dynamic range with somewhat lower sensitivity than with other dynamic ranges Low range on the other hand produces
84. e any precipitation or clotting or that contain any mechanical particles with the automatic dispensers A You may need to adjust the dispensing speed The default setting is for water You can find the adjustments and selections in SkanIt Software for Varioskan Flash Thermo Scientific Varioskan Flash User Manual 79 Routine Operation Other functions When dispensing is started the liquid volume in the well should be less than half of the total volume for example the volume should be less than 200 pl in a typical 96 well plate Emptying To avoid waste of reagents you may wish to empty reagent that is in the tubing back into the bottles This may well be the case if expensive reagents are used To empty Execute gt Empty follow these steps 1 The aspirate tube can remain in the reagent bottle 2 Enter an estimate 800 pl in pl of the volume of reagent remaining in the tubing 3 Ifyou do not wish to empty close the Empty dialog Refer to the emptying instructions in the SkanIt Software for Varioskan Flash User Manual Cat no N06355 Dispenser washing The Varioskan Flash can be washed using SkanIt Software for Varioskan Flash or the control switches for priming the dispensers Wash the dispenser tubing if necessary Either select Execute gt Wash or use the Prime Empty control switch located on top of each dispenser Figure 2 5 and Figure 2 16 Refer to the washing instructions in the Skanlt Software for
85. e dispensing tip when necessary See Replacing a v v dispensing tip on page 99 Replace the dispenser syringe if necessary See Replacing a dispenser v syringe on page 100 Ensure proper shutdown See Shutdown on page 84 v v Decontaminate the instrument when relocating the instrument or v sending it for service See Decontamination procedure on page 102 v Service the instrument regularly See Regular and preventive maintenance on page 88 Maintaining a system log on page 107 and Service contracts on page 108 Regular and preventive maintenance A 88 Thermo Scientific Varioskan Flash User Manual v depending on the laboratory conditions and the use and configuration of the instrument For reliable daily operation keep the instrument free of dust and liquid spills Abrasive cleaning agents are not recommended because they are likely to damage the paint finish It is recommended to clean the case of the instrument periodically to maintain its good appearance A soft cloth dampened in a warm mild detergent solution will be sufficient Caution Painted surfaces can be cleaned with most laboratory detergents Dilute the cleaning agent as recommended by the manufacturer Do not expose painted surfaces to concentrated acids or alcohols for prolonged periods of time as damage may occur A Plastic covers and surfaces can be cleaned with a mild laboratory detergent or ethanol It is
86. e input tubing incl tubing 3 port valve and end weight Dispenser syringe 1 ml and plunger Plunger lock screw Complete dispensing tube assembly i e the output tubing Dispensing head Dispensing tip 2 options 0 40 mm and 0 25 mm The tubing positions depend on the flow direction of the dispenser Figure 2 14 Varioskan Flash dispensing system There are two different dispensing tip options the black dispensing tip 0 40 0 40 mm and the transparent dispensing tip 0 25 0 25 mm Figure 2 15 Dispensing tip 0 25 Cat no N03081 has to be ordered separately and is intended for small volumes in the range of 1 to 20 pl Dispensing tip 0 40 Cat no N03080 is the default tip and is recommended for volumes gt 5 ul since the dispensing is then more rapid 30 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Control switches DISPENSER PRIME EMPTY Thermo Fisher Scientific control switch Incubator Functional Description Control switches Figure 2 15 Dispensing tip options 0 40 mm and 0 25 mm Refer to How to setup the dispensers on page 49 There are four white rocker switches one Plate In Out rocker switch for driving the plate carrier in or out Plate In Out function Figure 2 16 and three Prime Empty rocker switches for priming and emptying the dispenser tubing Prime Empty function of each dispenser Figure 2 16 Note that the Prim
87. ectors are inserted all the way and the connector locks are properly closed Figure 3 41 Connecting the cables 9 Mount the connector cover shield 54 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Thermo Fisher Scientific Installation Installation setups 10 Mount the Varioskan LumiSens optics vertically downwards to the Varioskan LumiSens base Figure 3 42 and lock the units by using the green lock knob Lock knob Figure 3 42 Mounting the Varioskan LumiSens optics 11 Ensure that the dispensing plugs are inserted into the dispensing positions on the Varioskan LumiSens base 12 Switch on the instrument The Varioskan LumiSens module installation is ready To set up the Varioskan LumiSens in SkanIt Software for Varioskan Flash follow these steps 1 Start SkanIt Software for Varioskan Flash 2 Connect to the instrument 3 When the connection is created the system will automatically identify the Varioskan LumiSens module and start the Varioskan LumiSens calibration procedure if there has either never been a LumiSens module connected before or if the instrument has been used without the Lumisens module the last time When the calibration does not start automatically refer to Step 6 for the instructions on manual calibration 4 Enter the Varioskan LumiSens serial number indicated on the right hand side of the module The serial number format is Lxxxx xx for example L0614
88. ects caused by poor workmanship for a period of one year from the date of delivery Thermo Fisher Scientific will repair or replace defective parts or materials during the term of warranty at no extra charge for materials and labor provided that the products were used and maintained in accordance with Thermo Fisher Scientific s instructions The warranty is invalid if products have been misused or abused For the warranty to be effective the product must have been purchased either directly from Thermo Fisher Scientific or from an authorized Thermo Fisher Scientific distributor The guarantee is not transferable to a third party without prior written approval from Thermo Fisher Scientific This guarantee is subject to the following exclusions e Any defects caused by normal wear and tear e Defects caused by fire lightning flood earthquake explosion sabotage war riot or any other occurrence of the type listed above e Refurbished products that are subject to different warranty conditions THIS WARRANTY IS IN LIEU OF ALL OTHER EXPRESSED OR IMPLIED WARRANTIES INCLUDING BUT NOT LIMITED TO ANY IMPLIED WARRANTIES OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE The seller is not liable for any loss or damage arising out of or in connection with the use of the product or other indirect damages These warranty terms and conditions can be obtained from your local Thermo Fisher Scientific dealer Consumables are not included in the warrant
89. ed to use the tip priming feature for accurate dispensing particularly when the dispensing volumes are small for example 1 to 20 pl For more information on the tip priming feature refer to Dispensing on page 76 All trays have a cavity for the tip priming vessel The holder for the tip priming vessel is located on the left side of the universal frame Figure 2 19 The robotic tray has two optional locations for the tip priming vessel depending on the robotic 34 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Tip priming vessel Position calibration hole s Universal tray with adapters Universal tray Universal frame adapter for incubating and measuring 96 384 and 1536 well plates or adapter for measuring 6 to 48 well plates A Thermo Fisher Scientific Functional Description Tray options configuration of either portrait position A or landscape orientation position B Figure 2 21 The tip priming vessel Figure 2 19 is an 8 well plate strip 1 x 8 Thermo Scientific Microtiter Solid Strip Assembly There are three holes for the automatic calibration and alignment of the track mechanism The hole marked Figure 2 19 is used for the positional check of the track mechanism at startup and when changing the tray This section provides information on the more commonly used universal tray and the adapters one can use with it Incubation can be carried out with 96 to 15
90. ell Refer to Photometric measurement module on page 28 e The fluorometric bottom reading optics directs the excitation and emission light from the spectral scanning module to measure fluorescence at the bottom of the microplate Refer to Fluorometric bottom reading on page 28 e The sensitive luminometer optics measures light produced in the microplate well Refer to Varioskan LumiSens measurement module on page 28 24 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Thermo Fisher Scientific Excitation optics Functional Description Optical system The excitation optics Figure 2 7 consists of the light source and the wavelength selection devices Warning Do not open the optical covers under any circumstances There is a risk of ultraviolet radiation injury Only authorized service personnel has permission to open the optical covers A Diffraction Kamen order 1 Mono 2 Mono Bandwidth Sarinah fagi selection chromator chromator selector xcitation light mae filters Figure 2 7 Excitation optics Light source A xenon flash lamp is used as the light source The lamp provides a wide spectral range needed for photometry and fluorometry The lamp is pulsed at a 100 Hz rate and activated only when measuring A short light pulse enables accurate TRF measurements One measurement consists of 1 to 1000 flash pulses according to the measurement quality and measurement speed requireme
91. ent parameters TRF delay time and TRF integration time The TRF delay time defines the time difference between the excitation flash and emission signal collection while the TRF integration time defines the time used for emission signal collection When the Varioskan Flash performs a TRF measurement it excites the sample with a very short light pulse waits for the defined TRF delay time and then collects the signal during the defined TRF integration time These actions form one TRF measurement cycle Figure 4 47 that is performed within a 10 ms period The cycle is repeated as many times as defined by the measurement time RFU Background fluorescence y J 7 J Time us Lamp Delay Measurement flash time time Figure 4 47 Structure of the TRF measurement cycle 6 Ifnecessary set the settle delay in SkanIt Software For more details refer to Settle delay on page 73 Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 69 Routine Operation Fluorometric spectrum scanning A A Fluorometric spectrum scanning Photometric measurement 70 Thermo Scientific Varioskan Flash User Manual Caution Do not open the measurement chamber door Figure 3 32 during measurement because this aborts the measurement A Caution Because of the relative nature of fluorometry it is recommended to use known samples or controls to verify instrument operation A The phases of the fluorometric spect
92. eration Emergency situations 86 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Chapter 5 Maintenance Maintenance This chapter on maintenance contains an outline of the points checklist mentioned in the checklist below Table 5 7 Contact local authorized technical service or your local Thermo Fisher Scientific representative for assistance if necessary Table 5 7 Maintenance checklist Item Weekly Monthly Yearly Keep the instrument free of dust See Regular and preventive maintenance on page 88 lt SN Daily Avoid disturbing any of the optical system components including optical covers and the reference chip Figure 2 18 See How to clean the measurement chamber on page 89 Wipe away spilled saline solutions solvents acids or alkaline solutions v from outer surfaces immediately to prevent damage and wipe with deionized distilled aqua See Regular and preventive maintenance on page 88 If any surfaces have been contaminated with biohazardous material v disinfect with a mild sterilizing solution See Regular and preventive maintenance on page 88 Clean the case of the instrument and the reagent basin periodically See Regular and preventive maintenance on page 88 Check the cleanliness of the measurement chamber monthly See How to clean the measurement chamber on page 89 Clean the tray when necessary See How to clean
93. eseeseeeeneeaeees 6 Chapter 1 Introduction to the Varioskan Flash ssssssssssseseessseseseeseseaeeees 15 OVERVIEW Js sheniasenceee a a a ea 15 nteended Useins newness caminiaenganirabeaains wd dinmasiaAseancadied aaa iria 16 Advantages of using Varioskan Flash esssessssseseeeeeeenceseeeeneeaees 16 Chapter 2 Functional Description sccsssscssssssssssessesessesssnssneeeestessenssneensaeeatens 19 Instr ment layouts ennonn a R R aE 19 Front Viewin a a A Aa a a a a aah 19 RTA A A EEE ENAT E A O E 19 Internal view sesssseesseessresseesseessrresrresserssttssteeseresseersseessressreeseeseees 20 Measurement techniques ssesesesesesssssssssrssesesessrseteterrerrerererersesees 21 Fluorescence intensity sssssessesesssetsesesssstststststssesestseesesesssesesene 21 Time resolved Mii resCence sa sox Laduicdsdesa sds tsshuded cassia totsodpucandautsboncos 21 Absoibancer nalne a A AA 22 Ta IN ES COT COs nania a a a a eaa a a heseee 22 Optical syste s aean th ee ei A eed 23 Principle of the optical system 2 210 840 cua Phat ca daa 24 Excitation Optiese oai aiie aae rE E i en 25 Measurement Optics sssessssessssesesesreresrseseseseererestsrseseeteresteeesee 26 Emission reading module sesseeeeeeeee seseris sesesesesesesesesee 27 Photometric measurement module sssseseseeeseeesreesereesresseeesre 28 Fluorometric bottom reading ssssesesesesesesesesesersrsesrerereeeereseseseses 28 Varioskan LumiSens me
94. ess Tel Fax PURCHASED FROM Distributor Address Tel Date of delivery Internet home page Date of purchase Your application area D O M O Mrs O Ms O Job title Position Surname block capitals First name block capitals Internet e mail address Excellent Above expectations Instrument installation o o Ease of use o o Flexibility oO oO User manual o o Software o o Customer support oO oO Overall o o As expected Below expectations Comments O O OOooaao OOoOdoagao Additional instrument and or software features desired Did you encounter any problems Where did you first learn about the product Would you like to receive information about other Thermo Scientific products Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 141 Thermo Scientific Varioskan Flash Feedback Form 142 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Appendix D Preparation of Luminometric Alignment Plate Required These required reagents materials and equipment are not provided by Thermo Fisher Scientific unless otherwise indicated reagents materials and Reagents equipment e ENLITEN ATP Assay System 100 assays Promega Corporation prod no FF2000 e Storage conditions prior to reconstitution the rL L Reagent and Reconstitution Buffer must be stored at 20 C Store the ATP Standard at 20 C 2 Dis
95. et 29 The dispenser is not responding Contact service 30 Non volatile parameters lost Contact service 31 The D A converters check of the A D converter board failed Contact service 32 The requested measurement method is not available Contact service 33 Dispensing or priming was attempted when there was no plate Insert a plate into the tray inserted into the tray 34 Dispenser tip priming was attempted when there was no tip Insert the tip priming vessel priming vessel inserted into the tray 35 This is only reported in response to the factory calibration Contact service 36 This is only reported in response to the factory calibration Contact service 37 This is only reported in response to the factory calibration Contact service 38 This is only reported in response to the factory calibration Contact service 39 This is only reported in response to the factory calibration Contact service 40 This is only reported in response to the factory calibration Contact service 41 This is only reported in response to the factory calibration Contact service 42 This is only reported in response to the factory calibration Contact service 43 This is only reported in response to the factory calibration Contact service 44 This is only reported in response to the factory calibration Contact service 45 XY table position calibration failed Contact service 46 No factory calibration for the current measurement method Contact service
96. expire period is 10 minutes Recalibration is performed 7 5 minutes after the previous calibration depending on the measurement procedure If a measurement is performed 10 minutes after the previous calibration a warning appears about calibration validity This may happen if an assay takes a long time to execute and there is no sufficient time slot to perform calibration during protocol execution However the warning does not mean that the measurement failed only that the accuracy of the results may have suffered In luminometric calibration a dark level control measurement is also performed to compensate for possible electronic component and photomultiplier signal level drift Fluorometric In fluorescence intensity FI or time resolved fluorescence TRF measurement measurements the following actions are carried out by the instrument 1 The tray is driven in 2 Excitation and emission wavelengths are selected by rotating the excitation and emission gratings 3 In the signal level calibration procedure the instrument reads the fluorescence from the reference chip Figure 2 18 compares it to the value in non volatile memory and sets a factor to correct the reading In long measurement procedures calibration is performed in a suitable phase without disturbing the measurement timing The default calibration interval is 10 minutes 4 The dynamic range is selected by the user Figure 4 46 e AutoRange e Manual range e High range
97. g head tube as a dispensing tip installation tool Insert the fragile tip into the smaller hole of the tool and then install it into the small piece of silicone New dispensing tip 0 40 mm E Dispensing head tube aes w Dispensing tip to be discarded Figure 5 59 Replacing the dispensing tip B Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 99 Maintenance Periodic maintenance 3 Replace the dispensing head tube Figure 5 59 Fasten the parts by turning in the opposite directions than shown in Figure 5 58 Replacing a When you replace a dispenser syringe with a different volume refer to dispenser syringe he Skane Software for Varioskan Flash User Manual Cat no P yring N06355 If the instrument has the wrong information about the syringe used it will dispense incorrect volumes To replace a dispenser syringe of 1 ml Figure 5 61 or 5 ml follow these steps Male connector of the dispensing tube assembly Aspirate tube assembly i e the input tubing incl tubing 3 port valve and end weight Dispenser syringe 1 ml and plunger Plunger lock screw Complete dispensing tube assembly i e the output tubing Dispensing head Dispensing tip 2 options 0 40 mm and 0 25 mm The tubing positions depend on the flow direction of the dispenser Figure 5 60 Dispenser assembly 1 Remove the liquid from the dispenser syringe Figure 5 60 and from the t
98. g on 41 51 63 64 symbols 4 syringe 15 29 50 76 78 79 80 96 97 99 100 101 115 131 132 145 146 size 29 115 system log 88 106 107 T time resolved fluorometry 15 16 21 22 25 33 67 68 69 77 120 127 135 145 146 147 tip priming 30 34 35 38 62 76 78 79 87 115 126 vessel 34 35 38 62 76 78 79 87 126 vessel holder 34 38 44 98 volume 115 top reading 112 119 track mechanism 32 33 34 35 45 46 73 105 transmittance 146 147 transport 41 42 43 44 45 46 47 48 104 105 106 107 111 damage 41 42 locks 41 43 44 45 46 47 48 104 105 106 107 package 42 104 tray 33 34 35 37 38 39 48 49 91 120 127 145 composition 34 frame 34 35 38 49 66 holder 33 34 41 43 44 45 48 49 104 105 107 127 transport lock 44 104 options 33 48 65 positioning 64 TRF 15 16 21 22 25 33 67 68 69 77 120 127 135 145 146 147 bottom 77 delay 69 147 measurement 21 25 69 127 145 top 77 TR FRET 22 120 145 146 147 troubleshooting 3 125 129 type label 4 Thermo Fisher Scientific U universal tray 16 31 32 33 34 35 36 48 75 79 131 132 upgrade kits 132 USB 41 111 132 147 user interface 111 V Varioskan LumiSens 15 24 28 29 41 43 47 52 53 54 55 56 57 58 59 60 62 64 71 87 88 92 93 94 95 104 106 107 114 125 127 128 131 132 base 53 54 55
99. h controls all the instrument functions and provides data reduction as well as reporting functions Computer interface USB 1 1 2 0 compatible or Thermo Fisher Scientific RS 232C Measurement types Fluorescence intensity time resolved fluorescence photometry and luminometry all with spectral scanning Incubator Plate specific incubation tray Shaker Orbital shaking Dispensers Up to three optional dispensers with automatic dispensing position control Plate size Plate maximum dimensions Universal tray 127 8 mm W x 85 8 mm D x 23 5 mm H Robotic tray 128 6 mm W x 86 0 mm D x 18 0 mm H Thermo Scientific Varioskan Flash User Manual 111 Technical Specifications Performance specifications Pe rfo rmance This section provides the performance specifications for the relevant g measurement techniques and other instrument capabilities specifications Table 6 10 Fluorometry Performance specifications Fluorometry Light source Xenon flash lamp Wavelength selection Double excitation and double emission monochromators Excitation wavelength range 200 1000 nm Emission wavelength range 270 840 nm Excitation bandwidth 5 nm and 12 nm Emission bandwidth 12 nm Wavelength setting resolution 1 nm Wavelength accuracy 3nm Xenon flash lamp Lamp lifetime typically 10 flashes 10 96 well microplates using 100 ms integration time per well Sensiti
100. h dependent amount of light absorbed and an emission spectrum which shows the wavelength dependent amount of light emitted No two compounds have exactly the same fluorescence spectra thus making fluorometry a highly specific analytical technique One of the major advantages of fluorescence detection is high sensitivity This is important as relatively small changes in for example ion concentration in living cells can have significant physiological effects In addition of fluorescence being a versatile tool in cell biology biochemistry and molecular biology it is also a powerful technique for studying molecular interactions in analytical chemistry physiology photochemistry and environmental science Fluorescence resonance energy transfer FRET is a fluorescence intensity based measurement technique Two labels are required for FRET measurements donor fluorescent and acceptor either fluorescent or non fluorescent The emission spectrum of the donor needs to overlap with the absorption spectrum of the acceptor to allow the energy transfer to happen FRET allows homogeneous assay formats to be used in the detection of biological interactions The change in the intensity of the generated FRET signal can be related to specific biological events such as enzyme mediated cleavage of DNA or protein substrates protein DNA interactions and protein peptide interactions Time resolved fluorescence TRF is a special form of fluorescence i
101. he priming volume is 1 pl The pull back feature prevents liquid droplets from forming onto the dispensing tip between dispensings Therefore the liquid is pulled slightly inwards This function can be set in SkanIt Software for Varioskan Flash or it can be turned off 0 If set the dispenser automatically pulls back 1 to 10 pl reagent with the 1 ml syringe and 5 or 10 ul reagent with the 5 ml syringe into the syringe every time the instrument dispenses It is always recommended to use the pull back feature when using the 5 ml syringe The dispensing and measurement operations can be synchronized with an exact time interval on a well to well basis Thus the instrument supports simultaneous dispensing and reading enabling fast kinetic measurements from the very start of the reaction However to support simultaneous dispensing and reading the dispensers have to be in their correct positions Figure 4 50 To minimize the delay before the measurement place the dispensing head into the corresponding dispensing position Refer to Figure 4 50 and Table 4 5 For more information refer to the SkanIt Software for Varioskan Flash User Manual Cat no N06355 Measurement and dispensing in normal and filter mode luminometric measurements is fastest when you use 96 well plates When you use 384 well plates the crosstalk shield moves between dispensing and measurement which causes a small delay Caution Never use any liquids that can caus
102. her covers of the Varioskan Flash than the dispenser sliding cover Figure 2 2 or measurement chamber door Figure 2 2 while the instrument is plugged into a power source Never open the measurement chamber door while the instrument is busy when the LED indicator is orange You can push in the tray manually only when the instrument is switched off Never force a microplate into the instrument The Varioskan Flash is intended for laboratory research use only Observe proper laboratory safety precautions such as wearing protective clothing and following approved laboratory safety procedures It is recommended that Good Laboratory Practices GLP are followed to guarantee reliable analyses Preventative maintenance instructions should be followed closely to keep the instrument in the best condition for maximum reliability A poorly maintained instrument will not give the best results Thermo Fisher Scientific Contents ntended useisiin a a a a a a a a e ai 3 How to use this user manual ssssssseesssesssseseresesresseessressrresrrersrerssees 3 For more information sessessessseessressersseessreeseressrrsssersseessressereeseesseess 3 Safety symbols and markings s ssesesesesesssssesesessssssesrsrsrsrsesesssesesesesessse 4 Safety symbols and markings used on the Varioskan Flash 4 Warning and other markings used in the documentation 5 Instrument safety and guidelines for use seseeessseeees
103. ic Varioskan Flash User Manual Figure 3 38 Removing the dispensing DaSC ce ccceccceceescsesteteteseetenesees 53 Figure 3 39 Releasing the green lock KnOb n ssnisesisenisenisinsinenineninenensn 53 Figure 3 40 Removing the connector protection shield ccc 54 Figure 3 41 Connecting the cables 0 0 cccccccccccsceeseeteseeteeteesteeeeeeeees 54 Figure 3 42 Mounting the Varioskan LumiSens OptiCs c ccccececeeee 55 Figure 3 43 Pulling up the filter lift cc eececeesesteceteteseseenenenees 59 Figure 3 44 Pressing down the filter into the filter wheel cee 59 Figure 4 45 Microplate loaded ccc ccc cecesesescetesesescetesesesteteseseaseesees 65 Figure 4 46 Dynamic range selection oo ccccccecececesescetetesesteteseseetetesees 68 Figure 4 47 Structure of the TRF measurement Cycle 69 Figure 4 48 ON OFF period time and total shaking time 2 111051112511122 74 Figure 4 49 Speed amp diameter combinations for different plate formats 74 Figure 4 50 Identification of the dispensing tip position 0 eee 76 Figure 5 51 Front cover removed 00 0 ceeecececesesesceceseeesesesceesesesteseseseacenenees 90 Figure 5 52 Internal view of the measurement chambe c cccce 90 Figure 5 53 Opening the dispensing base for Cleaning ccccceceee 92 Figure 5 54 Removing the Varioskan LUMISeNS optics 951110511151111 93 Figure 5 55 Cleaning the Var
104. igure 2 12 Fluorometric bottom reading optics The Varioskan LumiSens module is an option that mounts in the front of the unit between the dispensers and is connected to the measurement position L For an overview of the module see Figure 2 13 Thermo Fisher Scientific Thermo Fisher Scientific Dispensers Functional Description Dispensers Varioskan LumiSens module Photo multiplier tube FE Measurement optics Shutter amp filter selector Reference system LumiSens optics Automatic cross talk shield 96 384 1536 Luminescence emission Figure 2 13 Varioskan LumiSens measurement module The Varioskan LumiSens module measurement optics collects with a wide angle emitted light that is produced in the microplate well and measured by a photomultiplier tube A crosstalk shield blocks light from adjacent wells and automatically sets the right aperture depending on the microplate format There are specific apertures for 96 384 and 1536 well plates If larger than 96 format wells are used the module will use the 96 aperture The shutter and filter selector contains positions for open blocked and eight configurable filters The blocked position is automatically used to eliminate background drifting of electronic components and the photomultiplier tube A reference system is used to compensate for photomultiplier gain drift Up to three optional dispensers Fig
105. ing compound by Bouguer Lambert Beer s Law Equation 2 Az eCd Equation 2 where A absorbance molar absorption coefficient I mol cm C concentration mol I d pathlength cm Luminescence Luminescence is the emission of light at visible wavelength by a substance Luminescence is caused by the movement of electrons from more energetic states to less energetic states In contrast to fluorometry no excitation light is required Luminescence can be caused by chemical or biochemical changes The excitation energy is thus produced by a chemical reaction There are many types of luminescence that can be identified according to the source of energy which excites the emission When the light energy emitted results from certain chemical reactions chiefly oxidations such as in the slow oxidation of phosphorus at ordinary temperatures the emission is called chemiluminescence When the luminescent chemical reaction occurs in a living system such as in the glow of a firefly the emission is called bioluminescence Bioluminescence is luminescence produced by living organisms and is thought to be a type of chemiluminescence Other examples of bioluminescence include glowworms deep sea organisms and various fungi and bacteria found on rotting wood or decomposing flesh Both luminescence types are detected by the instrument 22 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Optical system Thermo Fishe
106. into the OFF position 2 Remove the dispensing heads from the dispensing positions and place them into the dispensing head holders 3 Remove the cover of the dispensing base by opening the two screws Figure 5 53 Figure 5 53 Opening the dispensing base for cleaning 4 Clean the dispensing base using a soft cloth or tissue paper soaked in a mild detergent solution soap solution or 70 ethanol Do not use formaldehyde 5 Close the cover of the dispensing base 6 The whole dispensing base can be removed for cleaning by opening the two screws Figure 5 53 if necessary To clean the Varioskan LumiSens optics follow these steps Thermo Fisher Scientific Maintenance Regular and preventive maintenance 1 Switch off the Varioskan Flash by pressing the power switch Figure 2 2 on the left side panel of the instrument into the OFF position 2 Place the dispensing heads into the dispensing head holders 3 Remove the Varioskan LumiSens optics Figure 5 54 by unfastening the green lock knob Turn the lever and lift the optics off upwards Lock knob Figure 5 54 Removing the Varioskan LumiSens optics 4 Soak a lintfree tissue in lens cleaning solution or isopropyl alcohol 5 Wipe over the optical lens in a straight movement from inside to out Figure 5 55 Do not use any circular movements when cleaning the optics Caution Never touch the surfaces of filters or optical lenses with bare hands
107. ion sensor 50 77 78 head positioner 77 78 96 position 4 30 50 55 62 72 76 77 79 92 111 119 speed 79 115 127 system 30 tip 30 31 51 62 76 79 81 88 97 98 99 107 115 131 tip cap 51 62 tip position 76 tip size 115 tube assembly 88 131 volume 34 38 78 115 disposal of instrument 108 disposal of materials 102 Do s and Don ts 61 62 63 Thermo Scientific Varioskan Flash User Manual 149 double monochromator 26 64 113 114 145 146 dynamic range 27 67 68 71 72 112 114 127 145 E emission 21 22 23 24 26 27 28 64 67 69 71 112 114 116 119 125 145 146 147 bandwidth 112 114 detector 27 diffraction order filters 27 64 125 monochromators 27 112 optics 24 27 28 71 wavelength range 112 emptying 31 80 103 107 end weight 132 environmental requirements 41 43 error 3 64 106 125 126 127 128 145 codes 125 messages 106 145 excitation 21 22 24 25 27 28 64 67 69 70 112 119 125 145 146 147 bandwidth 64 112 125 optics 24 25 27 28 wavelength range 112 F Feedback Form 141 FIA 147 fluorescence 15 16 21 22 24 28 33 67 68 69 111 112 119 120 135 136 145 146 147 intensity 15 16 21 22 33 67 68 69 111 112 119 120 135 146 147 lifetime 21 145 146 fluorochrome 145 fluorometric label 134 fluorometric measurement 23 24 26 67 70 72 fluorometric spect
108. ioskan LumiSens Optics cccccceeeeeeee 94 Figure 5 56 Cleaning the Varioskan LumiSens base from above 95 Figure 5 57 Cleaning the Varioskan LumiSens base from the bottom 95 Figure 5 58 Replacing the dispensing tip A 0 cccccccceceeeteeeteeeeteeee 99 Figure 5 59 Replacing the dispensing tip B c cece ce eseeeeteeeteees 99 Figure 5 60 Dispenser assembly occ cecececesesestetesesestesesesesteeseseees 100 Figure 5 61 Replacing the dispenser syringe c cece cetetesesteteeseees 101 Figure 5 62 3 port valve replaceMent ecececccecescscetesesesteesesesteteseseees 102 Figure 5 63 Transport lock released A and fastened B screws 1 4 ShOWN cccesccenstest detects aektycatdetsiatea eacsteatpslncateetecce stan eiauadeee 105 Figure 5 64 Varioskan LumiSens optics transport lock unfastened 106 Figure 5 65 Varioskan LumiSens optics transport lock fastened 106 Thermo Fisher Scientific Thermo Fisher Scientific Tables Tables Table 2 1 Compatibility of the universal tray and plate specific adapters 36 Table 2 2 Compatibility of the robotic tray and plate specific adapter 37 Table 3 3 Installation CheCKIiSt cece cecccecesecseseeseseeseeessesessesesseneeeeneees 41 Table 4 4 Front panel indicator light 2 00 ceeseeceteseseeteteeseeeeee 63 Table 4 5 Readings carried out in measurement positions F and L 77 Ta
109. ity of the robotic tray and plate specific adapter Measurement Incubation Dispensing yes 96 amp 384 no 1536 Description Robotic tray with adapter for plate without lid yes no To be used with the most common 96 to 1536 well plate formats without lids Incubation is not supported Dispensing is possible with 96 and 384 well plates 127 Robotic tray without adapter for yes no no plate with lid To be used with the most common 96 to 1536 well plate formats with lids Incubation and dispensing are not supported Robotic tray Robotic tray adapter for measuring and dispensing plates without lids or no adapter when measuring plates with lids Thermo Fisher Scientific The robotic tray comes equipped with the elevation adapter for plate without lid 126 and is thus directly ready for measurement of 96 to 1536 well plates and dispensing of 96 and 384 well plates Figure 2 20 Thermo Scientific Varioskan Flash User Manual 37 Functional Description Tray options Tip priming vessel in holder position A portrait orientation How to change the location of the holder for the tip priming vessel 38 Thermo Scientific Varioskan Flash User Manual Positioning lever Robotic tray frame Figure 2 20 Robotic tray fitted with adapter for plate w o lid 126 Remove the adapter when you use microplates with lids Refer to How to remove or replace the robotic tray adapter on page 39
110. le for achieving best sensitivity is to select the lowest possible range without receiving overrange results in the measurement The measured values are comparable regardless of the dynamic range selection AutoRange or any of the fixed manual ranges 7 The wells are measured with a selected measurement time that can vary from 10 to 10 000 ms The amount of used measurement time affects the quality of the measurement result Thus the more time the better the quality of the result It is recommended to measure using a 1000 ms measurement time If there is a necessity to improve the quality of the results the measurement time should be increased The result is the mean value of individual 10 ms readings during the total measurement time 8 If necessary set the settle delay in SkanIt Software For more details refer to Settle delay on page 73 Caution Ensure that the dispensing heads or the blind plugs are used in the dispensing positions prior to measurement A 72 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Luminometric spectrum scanning Settle delay Other functions Orbital shaking Thermo Fisher Scientific Routine Operation Luminometric spectrum scanning The phases of the luminometric spectrum scanning measurement are the same as for the luminometric measurement but it always uses the spectral scanning module monochromators for controlling the wavelengths Refer to Lumin
111. lters are used to block unwanted harmonic transmission from the monochromators Emission detector Emission light is converted into electrical signals by the photomultiplier tube PMT The dynamic range is adjusted automatically AutoRange default or manually according to the measurement situations The manual dynamic range setting has four selections Low Medium low Medium high and High range Figure 4 46 Thermo Scientific Varioskan Flash User Manual 27 Functional Description Optical system 28 Thermo Scientific Varioskan Flash User Manual Photometric measurement module Fluorometric bottom reading Varioskan LumiSens measurement module Besides fluorescence the emission optics is also used for generating luminescence spectra Photometric measurement is carried out by using the excitation optics module as the photometric measurement light source The photometric measurement module Figure 2 11 is located at the measurement position F Refer to Figure 2 6 Reference sensor Excitation light Photometer module Figure 2 11 Photometric measurement module Fluorometric bottom reading is based on fiber optics that transmits the light from the spectral scanning module to the fluorometric bottom reading optics Figure 2 12 The fluorometric bottom reading optics is located at the measurement position L Bottom measurement optics Reference sensor EX fiber link EM fiber link F
112. mber door Figure 3 24 Varioskan LumiSens To release the transport lock of the Varioskan LumiSens optics follow optics transport lock these steps 1 Unlock the gray Varioskan LumiSens optics transport lock by unfastening the two lower screws and releasing the top screw Figure 3 29 Locking screw fastened A z Figure 3 29 Varioskan LumiSens optics transport lock fastened 2 Turn the transport lock clockwise 90 and fasten the locking screws to their places of storage on the instrument Figure 3 30 Locking screw removed and stored Figure 3 30 Varioskan LumiSens optics transport lock removed Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 47 Installation Installation setups How to install the The universal and robotic trays are easy to install As a safety tray precaution the tray adapter combinations are individually coded for automatic identification Refer to Tray options on page 33 1 Remove the metallic yellow labeled transport lock of the tray holder Refer to How to release the transport locks on page 43 Lever opening bar Contact pins Screw fix hole Figure 3 31 Tray holder 2 Gently move the tray holder to the front of the instrument by hand Figure 3 31 3 Ensure that you have chosen the correct tray Refer to Tray options on page 33 4 When you install the tray into the instrument make sure the positioning lever is first p
113. minous solution into the well H12 on a white 384 well plate Select the correct plate from the list place the plate onto the plate tray and click Continue 6 The instrument searches for the optimal Varioskan LumiSens measurement position 7 When the alignment is ready the Varioskan LumiSens is ready for use Thermo Scientific Varioskan Flash User Manual 57 Installation Installation setups Varioskan LumiSens scaling factor Varioskan LumiSens module filter installation 58 Thermo Scientific Varioskan Flash User Manual The Varioskan LumiSens will use the aligned measurement position until the next position alignment is carried out Refer to the SkanIt Software for Varioskan Flash User Manual Cat no N06355 The Varioskan LumiSens module measurement values can be scaled to a certain signal level For example two instruments can be scaled to give the same signal level To establish the scaling factor a reference is needed This can be a user defined concentration or a calibration tool such as the Lumiwell verification plate Cat no 2806460 The reference signal is measured with the LumiSens optics and the scaling factor is manually calculated using the following formula Reference Value Scaling Factor __ _ LumiSens Value Where e Reference Value is the expected value of the reference e LumiSens Value is the measured value of the reference Enter the scaling factor on the LumiSens ta
114. n Enzymology 57 Academic Press New York 653 pages DeLuca M A and McElroy W D Eds 1986 Bioluminescence and chemiluminescence Part B Methods in Enzymology 133 Academic Press Orlando 649 pages Kessler C Ed 1992 Nonradioactive labeling and detection of biomolecules Springer Verlag Berlin 436 pages LaRosa R A Ed 1998 Bioluminescence Methods and Protocols Methods in Molecular Biology 102 Humana Press Totowa New Jersey 310 pages Mason W T Ed 1999 Fluorescent and Luminescent Probes for Biological Activity A Practical Guide to Technology for Quantitative Real Time Analysis Second edition Biological Techniques Series Academic Press Roda A Pazzagli M Kricka L J and Stanley P E Eds 1999 Bioluminescence and Chemiluminescence Perspectives for the 21st century John Wiley amp Sons Chichester U K 625 pages Ziegler M M and Baldwin T O Eds 2000 Bioluminescence and chemiluminescence Part C Methods in Enzymology 305 Academic Press San Diego 732 pages 136 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Appendix A System Log Instrument name number User Date Comments Thermo Fisher Scientific PHOTOCOPIABLE Thermo Scientific Varioskan Flash User Manual 137 System Log 138 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific
115. n its protective plastic wrapping until the ambient temperature has been reached Unpack the Varioskan Flash instrument and accessories carefully with the arrows on the transport package pointing upwards Refer to the enclosed packing instructions The following notes and instructions are sent with the instrument and are immediately available when you open the package e Packing instructions e Packing list e Warranty Certificate card e Performance test reports e Varioskan Flash User Manual and Quick Reference Guide e SkanIt Software for Varioskan Flash package Caution Do not touch or loosen any screws or parts other than those specifically designated in the instructions Doing so might cause misalignment and will void the instrument warranty A Retain the original packaging for future transportation The packaging is designed to assure safe transport and minimize transit damage Use of alternative packaging materials may invalidate the warranty Also retain all instrument related documentation provided by the manufacturer for future use If you relocate your instrument or ship it for service refer to How to pack for service on page 107 Check the enclosed packing list against order If any parts are missing contact your local Thermo Fisher Scientific representative or Thermo Fisher Scientific Oy Visually inspect the transport package the instrument and the accessories for any possible transport damage If the carton ha
116. nd Figure 3 36 The instrument recognizes a dispensing head placed into the dispensing position Dispensing will not occur if the instrument does not recognize that the dispensing head is in the dispensing position that is the dispensing head positioner The controller can consequently detect which dispensing head is in which dispensing position The controller can also detect whether the tip assembly is properly inserted or not Refer to How to setup the dispensers on page 49 When the dispensers are not in use the dispensing heads can be stored in the dispensing head holders Figure 2 5 The tip holes to the measurement chamber must be closed with blind plugs to prevent light from entering the measurement chamber Figure 2 5 and Figure 5 52 Caution To avoid stray light use blind plugs to close empty dispensing positions A Thermo Scientific Varioskan Flash User Manual 77 Routine Operation Other functions 78 Thermo Scientific Varioskan Flash User Manual Priming A A Tip priming The Varioskan Flash can be primed using SkanIt Software for Varioskan Flash or control switches for priming the dispensers Prime the dispenser tubing if necessary Either select Execute gt Prime or press the button on the toolbar in SkanIt Software for Varioskan Flash or use the Prime Empty control switch located on top of each dispenser Figure 2 5 and Figure 2 16 To prime remove the dispensing heads Figure 2 5 f
117. neenes 131 Vatigskan Fl s heeren one Celene ates aie Saati St 131 List of spare parts and accessories vzsd sat saedouesuheseesans eh oes saeedes 131 Upgrade kitsi sisii icis iesi aiseta aE e a 132 Refor entes a a a aaa aAa aaraa NA aaaeaii 133 Usehuileweb links scstesaiactenasscapetaieietviupd n RRR 133 Te Fe CU aoe te ce la 2s eed sae OS ae Seca Tae ce ed ate ree 135 Fluorescence intensity savicostensaissccssdeasancvaiewuevevniauacaaeesarecaeansaients 135 Time resolved fluorescence s essssesssseseseesesessssesesetersesessesesesee 135 Absorbance sedan esuestene tes R E E RRR R a 136 L minescenten in anen ee a a E a S a 136 System Log zenin 137 Certificate of Decontamination ssssss11111111rrrrsnsunnnnnnnnnnnnnnnnnnnenennnnnnns 139 Thermo Scientific Varioskan Flash Feedback FOorm ssss 141 Preparation of Luminometric Alignment Plate 143 Required reagents materials and equipment 143 Product COMPONE NES cic cs cated Rann nea ee Guten 143 Reconstitution of assay COMPONENLS 00 1 ceeseeeeseeeeecteeseeeeeeseeeeeees 144 Pipetting INSPECTION Su cece adi au tater aneiysatoani sa anutitaiataatSecctdauAnoatnns 144 Thermo Fisher Scientific Thermo Fisher Scientific Figures Figures Figure 1 1 Varioskan Flash spectral scanning multimode readet 15 Figure 2 2 Varioskan Flash front ViQW cccccccscescecesesestetesesesteteseseeteesees 19 Figure 2 3 Varioskan Flash back ViQW 19 Figure
118. ng to Decontamination procedure on page 102 Warning Ensure that the bottom of each microplate is dry Fluid on the bottom of a microplate may present a contamination hazard Use proper laboratory practices when handling any hazardous materials A Caution Keep all the holes in the adapter clean both the identification holes Figure 2 19 and the well holes Table 2 1 and Table 2 2 a Caution Do not autoclave the tray Figure 2 19 and Figure 2 20 A To clean the reagent basin and dispensing area follow the instructions below Caution Keep the reagent basin Figure 2 5 and dispensing area Figure 3 34 clean Clean the surface daily using a soft cloth or tissue paper soaked in a mild detergent solution soap solution or 70 ethanol Wipe up spills immediately Do not use formaldehyde A Thermo Scientific Varioskan Flash User Manual 91 Maintenance Regular and preventive maintenance A How to clean the dispensing base Open the two screws for removing the cover Open the two screws for removing the dispensing base How to clean the Varioskan LumiSens optics 92 Thermo Scientific Varioskan Flash User Manual Warning If any surfaces have been contaminated with biohazardous material a mild sterilizing solution should be used A To clean the dispensing base follow these steps 1 Switch off the Varioskan Flash by pressing the power switch Figure 2 2 on the left side panel of the instrument
119. ntensity where fluorescence lifetime of the signal is remarkably longer than in fluorescence intensity TRF uses lanthanide labels which have similar excitation and emission spectra as fluorescence intensity labels Every TRF label has a unique fluorescence lifetime parameter T tau which reflects the duration of fluorescence emission after excitation has been switched off In TRF measurements the lanthanide label is excited with a light flash and the resulting emission is detected after a label specific delay time Thermo Scientific Varioskan Flash User Manual 21 Functional Description Measurement techniques Typical biological samples have a fluorescence background with a very short lifetime which has an effect on fluorescence intensity measurements In TRF technology this biological background has decayed before the TRF signal is measured giving improved assay performance Time resolved fluorescence labels can well be used for resonance energy transfer applications as fluorescence intensity labels This time resolved fluorescence energy transfer technology is known as TR FRET Absorbance When a beam of light enters a sample part of the light is absorbed by the sample and the rest is transmitted passes through the sample Absorbance A is defined by Equation 1 A log I I Equation 1 where J intensity of incident light I intensity of transmitted light The absorbance is linearly related to the concentration of the absorb
120. nto account both the environmental and technical prerequisites Refer to Environmental requirements on page 43 and Things to avoid on page 43 L __ Install the instrument Refer to Installation setups on page 43 Release the transport locks of the tray holder and the Varioskan LumiSens module Refer to How to release the transport locks on page 43 Install the tray Refer to How to install the tray on page 48 Install the complete dispensing tube assembly if required Refer to How to setup the dispensers on page 49 J Install Skanlt Software for Varioskan Flash Refer to Installation of Skanlt Software for Varioskan Flash on page 51 Refer to the Skan t Software for Varioskan Flash User Manual Cat no N06355 Connect the mains supply cable and USB or serial RS 232C cable Refer to Cable connections on page 51 Perform the operational check Refer to Operational check on page 64 Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 41 Installation What to do upon delivery What to do upon delivery How to unpack Checking delivery for completeness Checking for damage during transport 42 Thermo Scientific Varioskan Flash User Manual This section covers the relevant procedures to be carried out upon arrival of the instrument Move the packed instrument to its site of operation To prevent condensation the instrument should be left i
121. nts Diffraction order selection filters Excitation diffraction order filters that is cut off filters are used to block unwanted harmonic transmission of monochromators The correct diffraction order filter is selected automatically Monochromators The monochromator is based on the diffraction grating A grooved surface of the grating diffracts the different colors into different angles and a bandpass wavelength is selected by rotating the grating Figure 2 8 The final pickup of the desired wavelength band is made by an entrance exit slit combination Two monochromators are serially connected for high spectral quality and this essentially minimizes leakage of undesired wavelengths that is stray light Thermo Scientific Varioskan Flash User Manual 25 Functional Description Optical system 2 rotating concave grating S A Faint stray light Input slit Faint stray light Gracon s diffraction Intermediate slit Exit slit Diffracted light amp faint stray light Selected pure wavelength ir co al 1 rotating concave grating Figure 2 8 Principle of the double monochromator Bandwidth selector The bandwidth is set by means of the monochromator slit width A selection of two bandwidths 5 nm and 12 nm is made by controlling the slits Measurement optics The measurement optics module Figure 2 9 is the front surface mirror optics system to generate a wavelength independent high definition beam
122. o install the Varioskan LumiSens module follow these steps 1 Remove the Varioskan LumiSens module from the package 2 Check for any damage 3 Ensure that the instrument is switched off 4 Dismount the dispensing base from the instrument by loosening the two screws with the hexagonal key that is included in the Varioskan LumiSens package Figure 3 38 The hexagonal key can be stored using the holes on the dispensing head holder to the right Thermo Fisher Scientific Installation Installation setups Unfasten the screws Figure 3 38 Removing the dispensing base 5 Dismount the Varioskan LumiSens optics from the Varioskan LumiSens base by unscrewing the green lock knob on the top of the unit and separating the optics from the base Figure 3 39 I lt Lock knob Figure 3 39 Releasing the green lock knob 6 Dismount the connector cover shield next to the Varioskan LumiSens base on the left Figure 3 40 Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 53 Installation Installation setups Figure 3 40 Removing the connector protection shield 7 Mount the Varioskan LumiSens base to the Varioskan Flash unit where the dispensing base was mounted using the same screws 8 Connect the Varioskan LumiSens cables to the connectors that are exposed when the cover shield was removed Figure 3 41 Start with the larger connector that fits the rear connector Ensure that the conn
123. ometric measurement on page 71 When liquid in the well is exposed to acceleration or deceleration surface resonance waves occur in the wells As the plate moves fast from one well and stops at the next well prior to a measurement the surface waves start propagating in the liquid The propagation continues for a certain time depending on the liquid and the well size The surface waves may affect the results and thus it is necessary to ensure that certain actions are taken to optimize the measurement The surface wave effect can be seen as noise in the signal in certain cases There are two methods to minimize surface wave effects when they occur 1 Use detergent in the well if possible 2 Increase the Settle delay time and wait until the surface waves have been sufficiently reduced For example in water based samples the surface waves may keep propagating up to e 5 seconds in a 6 well plate e 300 ms in a 96 well plate e lt 20 ms in a 384 well plate For more information on the settle delay refer to the SkanIt Software for Varioskan Flash User Manual Cat no N06355 The Varioskan Flash also has shaking incubating and reagent dispensing capabilities which are presented below The orbital shaking function is used for shaking the microplate to mix the samples Movement of the track mechanism Figure 2 18 can perform the shaking action Refer to Track mechanism on page 32 The shaking action can be intermittent
124. on 119 quantum yield 146 Quick Reference Guide 42 reagent 15 16 30 50 57 73 78 79 80 87 91 121 145 basin 87 91 bottle holder 30 reference chip 62 67 87 90 146 reflection grating 25 27 145 146 replacing a dispenser syringe 88 99 replacing a dispensing tip 88 98 replacing the aspirate tube assembly 88 98 robot 33 robotic 32 33 34 37 38 39 48 79 120 131 tray 32 33 34 37 38 39 48 79 131 rocker switch 31 S scaling factor 56 57 58 scanning 114 optics 114 screw fix 34 48 104 sensitivity 21 68 71 72 112 114 119 service 25 27 42 61 62 64 85 87 88 89 101 104 106 107 108 125 126 127 128 129 131 contracts 88 108 pack for 42 89 107 129 request protocol 89 129 setting range 114 settle delay 68 69 70 72 73 shaker 111 115 shaking 15 16 32 73 74 115 diameter 115 method 115 speed 115 shutdown 62 84 88 spare parts 131 specifications 3 16 111 general 108 111 performance 112 113 114 115 safety 108 115 116 spectral scan 15 16 24 28 61 73 111 112 113 114 spectral scanning 15 16 24 28 61 73 111 112 113 114 152 Thermo Scientific Varioskan Flash User Manual spectrophotometric verification plate 122 132 speed 68 72 74 98 112 113 114 127 starter reagent 77 startup 32 35 63 64 122 126 127 step 77 119 Stokes shift 146 stray light 4 25 62 77 113 switchin
125. ormed regularly and on a daily basis to ensure proper dispenser operation 1 Rinse the dispenser tubings Figure 3 36 out thoroughly with distilled water after each use Leave the fluid pathway filled for storage 2 Inspect the dispensers for leaks and correct any problems immediately 3 Wipe up all spills on and around the dispensers immediately 4 Do not allow the dispensers to run dry for more than a few cycles Clean the fluid path thoroughly on a weekly basis to remove precipitates such as salts eliminate bacterial growth and so on using one of the procedures outlined below There are three agents with which the dispensers may be cleaned e Weak detergent e 10 bleach for example sodium hypochlorite e Weak base and acid Thermo Fisher Scientific Weak detergent or 10 bleach Weak base and acid in Thermo Fisher Scientific sequence Maintenance Regular and preventive maintenance Remove the dispensing heads Figure 2 5 from the dispensing head positioners Figure 2 5 and do not let any cleaning fluids enter the measurement chamber Figure 5 52 Use external containers To clean the dispensers Figure 3 34 with weak detergent or 10 bleach follow these steps 1 Prime the dispenser with a weak detergent solution or a 10 bleach solution Make a solution of 10 bleach by adding one part of commercial bleach to nine parts of water Leave the solution in the dispenser with the syringe Figure
126. pensing are possible with 384 well plates Recommended for reading of 1536 well plates 15 mm high plates in all measurement modes and 10 mm high plates in fluorometric mode yes 384 no 1536 5 384 well adapter for plate with lid yes yes no To be used with the most common 384 well plate formats with lids Incubation is possible No dispensing 65 1536 well adapter for plate w o lid yes yes no To be used with the most common 10 mm high 1536 well plate formats without lids in luminometric mode Incubation is possible No dispensing 80 6 48 well adapter for plate without lid yes no yes To be used with the most common 6 to 48 well plate formats without lids Incubation is not supported Dispensing is possible 48 6 48 well adapter for plate with lid yes no no To be used with the most common 6 to 48 well plate formats with lids Incubation and dispensing are not supported Continued 36 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Cont Picture Functional Description Tray options Description Measurement Incubation Dispensing ID no 126 Picture 96 well adapter for PCR plate without lid yes yes yes To be used with 96 well PCR plates with NUNC product codes 230012 264605 or equivalent plates from other plate manufacturers Incubation and dispensing are supported included in standard deliveries Table 2 2 Compatibil
127. posables e Pipette tips for pipetting 15 pl and 200 1000 pl volumes e Thermo Scientific White Microlite 1 Plate 384 well plate Cat No 8155 or any other solid white 384 well microplate e Disposable 2 ml or 2 5 ml Eppendorf tube or equivalent 3 Equipment e Thermo Scientific Finnpipette for 10 100 pl with 15 pl volume e Thermo Scientific Finnpipette for 200 1000 pl volumes Product o L LReagent 1 vial components e Reconstitution Buffer 12 ml e ATP Standard 10 7M 1 vial e ATP Free Water 25 ml not needed in this test Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 143 Preparation of Luminometric Alignment Plate Reconstitution of assay components Pipetting instructions rL L Reagent Before opening gently tap the rL L Reagent vial to ensure that the lyophilized material is in the vial bottom Slowly remove the vial crimp seal and rubber stopper to avoid loss of material Add the entire contents of the plastic bottle labeled Reconstitution Buffer to the vial replace the rubber stopper and gently swirl the vial to dissolve the contents DO NOT shake the dissolved rL L Reagent Allow the rL L Reagent to equilibrate to room temperature before use Reconstituted rL L Reagent can be held at room temperature for 8 hours If the reagent will be used for longer than 8 hours dispense the rL L Reagent into 200 1000 ul aliquots and store them at 4 C protected from light Use aliq
128. r Scientific Functional Description Optical system Bioluminescence resonance energy transfer BRET is a non destructive cell based assay technology that offers the ability to directly study complex protein protein interactions in living cells The assay is based on non radiative energy transfer between fusion proteins containing a luciferase bioluminescent donor and a green fluorescent protein GFP mutant fluorescent acceptor Interactions between the two fusion proteins can bring the luciferase and GFP close enough for resonance energy transfer to occur thus changing the color of the bioluminescent emission The transfer efficiency depends on the degree of the spectral overlap the relative orientation and the distance between the donor and acceptor BRET is also a naturally occurring phenomenon in marine animals such as the sea pansy Renilla reniformis and the jellyfish Aequorea victoria In most applications the fused donor is Renilla luciferase Rluc rather than aequorin to avoid any intrinsic affinity for Aequorea derived GFP mutant the acceptor is the yellow fluorescent protein YFP to increase the spectral distinction between the two emissions The Varioskan Flash uses fluorometric luminometric and photometric measurement techniques Fluorometric measurements are made from the top or bottom of the well luminometric measurements from the top of the well and photometric measurements through the well The principle of
129. r element is slightly warmer than the lower element to avoid condensation on the plate lid On the other hand heating can be carried out with all trays and adapters for example the robotic tray by using only the upper heater element located in the ceiling of the measurement chamber to minimize condensation on the plate lid Microplate 96 well adapter for plate without lid 2 Bottom reading hole Universal tray frame Figure 2 17 Varioskan Flash incubator cross section The track mechanism Figure 2 18 has been specifically designed to obtain excellent measurement results for different plate formats The position calibration hole on the top left corner of the tray is read at startup and plate positioning is adjusted accordingly Also if the tray is changed the calibration hole is read again The home sensors of the X and Y carriages are used for checking the correctness of the plate position The check is done each time the carriage passes the home position and also always when the plate is driven out Note that movement of the track mechanism can perform orbital shaking Refer to Orbital shaking on page 73 and Table 6 14 Thermo Fisher Scientific Reference chip Functional Description Tray options Y carriage X carriage Reference chip top reading bottom reading Tray options Thermo Fisher Scientific Tray holder Tray Figure 2 18 Part of the Varioskan Flash track mechanism
130. recommended below or some other corresponding decontamination procedure must be performed It is strongly recommended to perform the complete decontamination procedure before relocating the instrument from one laboratory to another 102 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Thermo Fisher Scientific gt Maintenance Decontamination procedure Decontamination is not required for the proper functioning of the instrument Example of decontaminants e Ethanol 70 e Virkon solution 1 3 e Glutaraldehyde solution 4 e Chloramine T e Microcide SQ 1 64 Caution If local or laboratory regulations prescribe regular decontamination it is not advisable to use formaldehyde since even small traces of formaldehyde negatively affect the enzyme being used in FIA tests resulting in bad test results A Warning The decontamination procedure should be performed by authorized trained personnel in a well ventilated room wearing disposable gloves protective glasses and clothing A To decontaminate the instrument follow these steps 1 Prepare the decontaminant 200 ml 4 glutaraldehyde solution or another agent recommended by your safety officer 2 Empty the tray Figure 2 19 and Figure 2 20 Ensure that you are wearing disposable gloves 3 Switch OFF the power Figure 2 2 and disconnect the mains supply cable Figure 3 37 4 Disinfect the outside of the instrument using a cloth dampene
131. rioskan LumiSens measurement position alignment Refer to the next section Note After the LumiSens calibration the signal level of the LumiSens module can change If a LumiSens scaling factor other than 1 has been used it is necessary to redefine the scaling factor to achieve similar signal levels than previously A Varioskan Flash has a function to optimize the luminometric measurement position The measurement position is determined based on the plate dimension information in the plate templates but the optimal position is somewhat different for each instrument because of individual mechanical variations between the units Optimizing the measurement position for each unit will ensure the specified crosstalk performance of the instrument The LumiSens module is fully functional and can also be used without this luminometric alignment but slightly increased crosstalk is to be expected with 384 and 1536 well plates When the alignment is run the instrument will search for the exact position of the defined well within the plate and store the correction information This correction will be used in all subsequent measurements and it will minimize luminescence crosstalk from Thermo Fisher Scientific Thermo Fisher Scientific Installation Installation setups adjacent wells in luminometry The measurement position alignment should be run after the installation of a new LumiSens module and when the LumiSens base has been removed for cleaning or
132. rom the dispensing head positioners Figure 2 5 and prime the dispensers by discarding the liquid into an external waste container Priming can also be carried out with the dispensing head in the dispensing head holder and with a small vessel underneath For safety reasons there must be a plate on the tray during priming Insert an empty plate of the same type as the actual assay plate into the tray The dispensing head position sensor will prevent priming if the dispensing head is located in the dispensing head positioner However the empty plate is used as a safety plate if the dispensing heads would be improperly inserted into the dispensing head positioners during priming Visually check that the dispensing jets are straight Finally insert the dispensing heads properly into the dispensing head positioners Refer to the priming instructions in the SkanIt Software for Varioskan Flash User Manual Cat no N06355 The minimum priming volume is 800 pl and the recommended volume 1500 ul Caution Carefully insert the dispensing head in one go into the dispensing head holder Otherwise the dispensing head can become damaged or contaminated A Caution Do not use priming vessels higher than the actual plate intended to be used in the assay A The instrument has a Prime tip ul feature see the SkanIt Software for Varioskan Flash User Manual Cat no N06355 This function can be selected in SkanIt Software for Varioskan Flash or it
133. rum scanning 70 fluorometry 16 21 22 24 25 66 70 112 119 146 147 frame 34 35 38 49 66 FRET 21 145 146 147 G glossary 145 grating 25 125 145 146 guide pin 38 48 guidelines for use 6 150 Thermo Scientific Varioskan Flash User Manual H heater 32 heating 32 75 High range 27 67 68 72 how to change the location of the holder for the tip priming vessel 38 how to clean the dispensing base 87 92 how to clean the measurement chamber 87 88 89 how to clean the optics in the measurement chamber 87 90 how to clean the tray 87 91 how to decontaminate the instrument 84 88 89 90 91 102 103 104 108 129 139 how to ensure startup 41 51 how to install the tray 33 34 41 48 65 how to install the Varioskan Flash 41 43 51 64 how to load the microplate 65 how to maintain the instrument 3 87 how to pack for service 42 89 107 129 how to refit the transport locks 104 107 how to release the transport locks 41 43 48 104 how to remove or replace the robotic tray adapter 37 39 how to shut down 84 88 how to switch on 63 64 how to unpack 41 42 incubation 15 16 32 33 34 35 36 37 75 111 120 incubator 31 32 48 64 75 111 114 121 installation 3 41 43 48 49 51 52 56 58 60 62 99 101 107 128 141 checklist 41 instrument layout 19 intended use 3 16 K kinetic measurements 16 79 123 L lamp 25 28 64 6
134. rum scanning measurement are the same as for the fluorometric measurement but with a continuous range of wavelengths Refer to Fluorometric measurement on page 67 In absorbance Abs measurements the following actions are carried out by the instrument 1 The tray is driven in 2 The measurement wavelength is selected by rotating the excitation gratings 3 In the photometric calibration procedure the instrument reads the air blank level In long measurement procedures calibration is performed in a suitable phase without disturbing the measurement timing The calibration is valid for 1 hour 4 The wells are measured with a selected measurement time that can vary from 10 to 1000 ms There is one xenon lamp flash for each 10 ms period of measurement time 5 The amount of xenon lamp flashes affects the quality of the signal Thus the more flashes the better the quality of the result The amount of flashes can be set to 1 to 100 flashes per measurement 10 1000 ms It is recommended to measure using a 100 ms measurement time default which produces good results If there is a necessity to improve the quality of the results the flash amount should be increased The result is the mean value of the number of 10 ms readings during the total measurement time Longer than 100 ms measurement times are recommended to reduce noise if the measured absorbance level is high 6 If necessary set the settle delay in SkanIt Soft
135. s been damaged in transit it is particularly important that you retain it for inspection by the carrier in case there has also been damage to the instrument Thermo Fisher Scientific Environmental requirements A Things to avoid Technical prerequisites Installation setups How to release the transport locks Thermo Fisher Scientific Installation Installation setups If any parts are damaged contact your local Thermo Fisher Scientific representative or Thermo Fisher Scientific Oy When you set up your Varioskan Flash avoid sites of operation with excess dust vibrations strong magnetic fields direct sunlight draft excessive moisture or large temperature fluctuations e Make sure the working area is flat dry clean and vibration proof and leave additional room for cables covers and so on e Make sure the ambient air is clean and free of corrosive vapors smoke and dust e Make sure the ambient temperature range is between 10 C 50 F and 40 C 104 F e Make sure relative humidity is between 10 and 80 non condensing The Varioskan Flash does not produce operating noise at a level that would be harmful No sound level measurements are required after installation Caution Do not operate the instrument in an environment where potentially damaging liquids or gases are present A Do not smoke eat or drink while using the Varioskan Flash Wash your hands thoroughly after handling test fluids Ob
136. s reported rpm Revolutions per minute self tests Initialization tests and adjustments that the instrument performs before operation as well as autocalibration Stokes shift The difference between the wavelengths of the excitation and emission peaks T tau Fluorescence lifetime of the TRF label Thermo Fisher Scientific Glossary transmittance transmittance The ratio of transmitted J and incident light 4 Wh tray The plate carrier into which the microplate is loaded for measurement and or dispensing TRF Time resolved fluorometry fluorescence Fluorescence intensity measurement using special labels TRF delay Waiting period between the excitation flash end and the beginning of the emission light measurement TR FIA Time resolved fluoroimmunoassay TR FRET Time resolved fluorescence resonance energy transfer USB Universal serial bus w With w o Without Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 147 Glossary 148 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Index T tau 21 145 146 3 3 port valve 12 101 102 132 A absorbance 15 16 21 22 70 113 120 136 145 146 accessories 13 42 50 131 accuracy 2 66 67 78 97 113 115 146 adapter 11 13 32 33 34 35 36 37 39 48 49 61 65 75 91 120 131 145 advantages of using Varioskan Flash 16 application 2 5 16 22 23 31 61 62
137. scence resonance energy transfer Refer to Measurement techniques on page 21 grating Reflective diffraction grating an arrangement of closely spaced reflecting grooves on a flat or concave surface Light can reflect on the grating but can also be diffracted An optical grating can contain a thousand lines or more per centimeter initialization tests So called self tests which are carried out before operation to ascertain that the necessary instrument adjustments have been carried out LANCE PerkinElmer s TR FRET kit family using Europium ACP LanthaScreen Invitrogen s TR FRET kit family using Terbium Fluorescein dual labeling LED Light emitting diode luciferase A generic name for enzymes commonly used in nature for bioluminescence luminescence Emission of light other than from thermal energy causes such as bioluminescence luminometer An instrument used for measuring the intensity of luminescent radiation luminometric label luminophore A substance which emits light at room temperature A group of atoms that can make a compound luminescent monochromator Transmits certain wavelengths of the incoming light Consists of a rotatable diffraction grating and entrance exit slits in this context multiplexing When two or more labels are used in the assay either simultaneously or consecutively e g in fluorometric FRET luminometric dual reporter gene and dual label TRF assays optical density
138. seereeeeeeeneeee 89 How to clean the optics in the measurement chamber 90 How to clean the ttaycccessexssasevaSysteicacceieessascascansclavteearertiaeaseaeaness 91 How to clean the reagent basin and dispensing area n se 91 How to clean the dispensing bases 2c2 re cscs taeciestieieaeets 92 How to clean the Varioskan LumiSens optics sscecseseeeeeereeee 92 How to clean the Varioskan LumiSens base eseeeseseeseeeeeeeeee 94 Routine maintenance of the dispensers ceeesesseeeeeceseeeeeeeeeeeees 96 Dily maintenancense 4s sewcncy sacastteiatyrantesecqas ina e oat 96 Weekly maintenance secscesssiirisiiennie anasi 96 Weak detergent or 10 bleach 97 Weak base and acid in sequence n se 97 Periodic maintenance sa lt stisrarencdisnsveasssdeetsaranceessadersondeatzedcisteaneytesdues 98 If the plunger isstutk sssini iseasi ais 98 Replacing the aspirate tube assembly or the complete dispensing tubeassembly nenei nie n R E E E E E 98 Replacing a dispensing tips tea Ge eee fe Ge 99 Replacing a dispenser syringe 26 c ceca 0S a ats Oaaa hayes 100 Replacing the 3 port valve scrcssccsiatssasvansytecnwsseytvioncwve dius Nosema edcas 101 Disposal of materials 8 sccdeecina catimsntyentlininncetmeiee 102 Decontamination Procedure se excise seed von otecnieseaecianeca a Menseserotentte 102 How to refit the transport locks 2 35 45 ocre eg avticed ecctuclenditedelse 104 Tray holder transport locke i563 temo iuaeas iets a ate alee 104
139. ser Manual 65 Routine Operation Switching on 66 Thermo Scientific Varioskan Flash User Manual Automatic runtime calibration 4 Load the microplate onto the instrument tray for measurement Figure 4 45 The tray is able to handle microplates of different sizes therefore the free space in the tray is slightly larger than for example the standard 96 well plate The positioning lever in the tray Figure 2 19 will automatically position the plate correctly into the upper left corner of the tray when the tray is driven in Always insert the microplate so that the Al corner is positioned in the top left corner of the tray Figure 4 45 The tray frame is marked with A1 to facilitate correct insertion of the microplate 5 After this the measurement is executed with SkanIt Software for Varioskan Flash The software measures according to the selected measurement parameters Refer to the SkanIt Software for Varioskan Flash User Manual Cat no N06355 Runtime calibration is always automatically performed at the beginning of the protocol execution The instrument also performs calibrations during protocol execution if it does not violate the timing requirements of the assay For example in a kinetic assay if a long enough kinetic interval time is defined so that there is time for calibration before each kinetic repeat then calibration can be performed between the repeats In photometry each selected wavelength is calibr
140. serve normal laboratory procedures for handling potentially dangerous samples Use proper protective clothing Use disposable gloves Ensure that the working area is well ventilated Never spill fluids in or on the equipment Place the instrument on a normal laboratory bench The net weight of the unit is 55 to 67 kg 121 148 lbs depending on the configuration The instrument operates at voltages of 100 240 Vac and the frequency range 50 60 Hz This section describes the installation setups that have to be carried out before instrument operation Two transport locks are present in the instrument Figure 3 23 one for the tray holder and another for the Varioskan LumiSens module Make sure the transport locks have been released before you put the instrument into operation Thermo Scientific Varioskan Flash User Manual 43 Installation Installation setups VARIOSKAN Fa ISH Figure 3 23 Transport locks and transport lock tags present Tray holder transport The tray holder transport lock support is easily recognizable since it has lock a metallic color and a yellow label Figure 3 26 To release the transport lock of the tray holder follow these steps 1 Lift up the dispenser sliding cover and also open the measurement chamber door slightly Figure 3 24 Lift up the dispenser sliding cover and open IN the measurement chamber door slightly Figure 3 24 Dispenser sliding cover and measurement ch
141. sh Varioskan Flash Cable connections This section shows the location of all relevant connectors and how to connect the mains supply cable Warning Ensure that the mains switch Figure 2 2 on the left side panel is in the OFF position Never operate your instrument from a power outlet that has no ground connection A Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 51 Installation Installation setups Varioskan LumiSens module installation 52 Thermo Scientific Varioskan Flash User Manual 1 Connect the mains supply cable to the mains power supply connector Figure 3 37 on the left side panel If you need to use any other type of mains supply cable than supplied use only cables certified by the local authorities 2 Connect the instrument mains supply cable to a correctly installed line power outlet that has a protective conductor that is grounded od USB connector Al lt SERIAL RS 232 C Serial connector 2 Mains power supply connector Figure 3 37 Connecting the mains supply cable 3 Connect the instrument to the PC using either the USB or serial communication cable The Varioskan Flash is field upgradable with the Varioskan LumiSens module The installation procedure consists of two parts 1 installing the Varioskan LumiSens module to the Varioskan Flash and 2 setting up the Varioskan LumiSens functionality in SkanIt Software for Varioskan Flash T
142. ssel Figure 2 21 by unfastening the holder retaining screw Figure 2 21 fitted with a washer 2 Place the holder for the tip priming vessel in the new position so that the guide pin fits in its hole Figure 2 21 The guide pin controls that the holder is placed correctly Then fasten the holder retaining screw fitted with a washer Position A Guide pins Holder for tip priming 4 vessel position A portrait si orientation position B landscape orientation Washer and retaining amp gt screw for holder for tip 22 gt priming vessel 7 Position B Figure 2 21 Changing the location of the tip priming vessel holder How to remove or The elevation adapter is needed for dispensing into microplates without replace the robotic tray lids Remove the adapter however when you use microplates with lids adapter To remove ot replace the robotic tray adapter Figure 2 22 follow these steps 1 To remove the factory installed adapter unfasten the four adapter retaining screws fitted with washers Figure 2 22 by turning them counterclockwise Keep the retaining screws and washers for future use by screwing them back onto the adapter 2 To replace the adapter Figure 2 22 first fasten loosely all four adapter retaining screws fitted with washers by turning them clockwise Then take a firm grip of the adapter and push the adapter towards the Al corner and fasten the Al corner adapter retaining screw firmly After
143. t an empty run to verify proper instrument operation 64 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Routine Operation Switching on Loading the This section describes what issues to take into account when loading a microplate microplate Caution Ensure that you select a correct plate type Too high a plate may get jammed and with too low a plate the dispensing might fail and ras pass by A Caution When placing a microplate onto the tray always make sure the correct plate type has been selected in SkanIt Software for Varioskan Flash Protocol Properties Plate template before you do anything else A To insert the microplate follow these steps 1 Ensure that the plate type tray adapter and the SkanIt Software for Varioskan Flash plate template match Refer to Tray options on page 33 and How to install the tray on page 48 Microplate Tray Adapter Plate template see Tray options on page 27 see the Skanlt Software for Varioskan Flash User Manual Cat no N06355 2 The tray is driven out after which the microplate can be loaded 3 Ifthe tray is in first drive the tray out by pressing the Plate Out rocker switch or by selecting either Execute gt Run Plate Out or clicking the SM button on the protocol action panel in SkanIt Software for Varioskan Flash Figure 4 45 Microplate loaded Thermo Fisher Scientific Thermo Scientific Varioskan Flash U
144. t techniques Guilbault G G Ed 1990 Practical Fluorescence Second edition Marcel Dekker Haugland R P Ed 2001 Molecular Probes Handbook of Fluorescent Probes and Research Products Eighth edition Molecular Probes CD ROM Lakowicz J R Ed 1999 Principles of Fluorescence Spectroscopy Second edition Plenum Pub Corp Mason W T Ed 1999 Fluorescent and Luminescent Probes for Biological Activity A Practical Guide to Technology for Quantitative Real Time Analysis Second edition Biological Techniques Series Academic Press Alpha B Lehn J M and Mathis G 1987 Energy Transfer Luminescence of Europium III and Terbium III Cryptates of Macrobicyclic Polypyridine Ligands Angew Chem Int Ed Engl 26 266 7 Biazzo Ashnault D E Park Y W Cummings R T Ding V Moller D E Zhang B B and Qureshi S A 2001 Detection of insulin receptor tyrosine kinase activity using time resolved fluorescence energy transfer technology Anal Biochem 291 155 158 Bookout J T Joaquim T R Magin K M Rogan G J and Lirette R P 2000 Development of a dual label time resolved fluorometric immunoassay for the simultaneous detection of two recombinant proteins in potato J Agric Food Chem Dec 48 5868 73 Halminen M Sj roos M M kel M J Waris M Terho E L vgren T and Ilonen J 1999 Simultaneous detection of IFN gamma and IL 4 mRNAs using RT PCR and time resolved fluorome
145. tall the syringe and pull the syringe plunger until it is above the carriage Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 101 Maintenance Disposal of materials 6 Align the valve using the plunger as a guide and tighten from 1 8 to turn after the syringe touch off 7 Pull the syringe plunger all the way into the carriage and secure by tightening the plunger lock screw Valve Tubing Ports Valve Plug 6 Y Valve Body Screw Mounts Valve Coupler Fitting Front View Syringe Mount Rear View Not shown Always located at bottom Figure 5 62 3 port valve replacement Disposal Of Follow laboratory and country specific procedures for biohazardous or radioactive waste disposal Refer to local regulations for the disposal of materials infectious material plates strips priming vessels disposable gloves syringes disposable tips and so on as biohazardous waste A A Warning The samples can be potentially infectious Dispose of all used Decontamination Decontamination should be performed in accordance with normal laboratory procedures Any decontamination instructions provided with procedure the reagents used should be followed A decontamination procedure is only recommendable when infectious substances have been in direct contact with any part s of the instrument If there is a risk of contamination with biohazardous material the procedure
146. tation and emission double monochromators e excitation and emission diffraction order filters e non volatile memory e temperature measurement electronics e measurement electronics e excitation bandwidth selector e xenon flash lamp e reference detectors e Varioskan LumiSens filter positioning e Varioskan LumiSens crosstalk shield positioning e dispensers e the measurement channel s dark level When the initialization tests and adjustments have been successfully completed the LED indicator Figure 2 2 turns from orange to green Refer to Switching on on page 63 If anything fails in the initialization tests or adjustments the LED indicator will turn red In this case try switching the instrument OFF and ON again If the failure is repeated contact authorized technical service All error messages are stored in the internal memory log file of Varioskan Flash The error log file can be accessed with SkanIt Software by selecting Settings gt Instrument gt Setup gt Reports gt Error Log gt Run Report Refer to Error and warning codes on page 125 The instrument also performs automatic signal long time stability checks during runtime After startup the instrument is ready for operation Since the instrument calibrates itself you can start measuring immediately as soon as the instrument has been turned on However the stabilization of the incubator can take up to 10 minutes It is further recommended to carry ou
147. ted Explanation Internal firmware error When this error occurs the internal software halts This error can only be seen in the instrument error log Suggested action Contact service error it is best to abort the current run and restart from the beginning after the problem is fixed The error Table 8 18 and warning codes Table 8 19 that may appear in SkanIt Software for Varioskan Flash 2 The instrument did not recognize the command it received Contact service 3 The arguments of the received command are not valid Contact service 4 The X carriage position is incorrect Contact service 5 The Y carriage position is incorrect Contact service 6 The 1 excitation grating position is incorrect Contact service 7 The 2 excitation grating position is incorrect Contact service 8 The 18 emission grating position is incorrect Contact service 9 The 2 emission grating position is incorrect Contact service 10 Excitation diffraction order filter position is incorrect Contact service 11 Emission diffraction order filter position is incorrect Contact service 12 Excitation selector position is incorrect Contact service 13 Emission selector position is incorrect Contact service 14 Varioskan LumiSens filter position is incorrect Contact service 15 Excitation bandwidth selector position is incorrect Contact service 16 Excitation beam diameter selector position is incorrect
148. that fasten the rest of the adapter retaining screws firmly to the tray Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 39 Functional Description Tray options Adapter for plate oa 12g BS without lid 126 VA Washer and adapter gt 8 retaining screw S2 Figure 2 22 Removing or replacing the adapter for plate w o lid 126 Refer to Table 2 2 and Chapter 9 Ordering Information 40 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Chapter 3 Installation Installation This chapter on installation contains an outline of the points checklist mentioned in the checklist below Table 3 3 depending on the configuration and care must be taken when lifting it A Warning The Varioskan Flash weighs 55 to 67 kg 121 148 lbs Two persons must lift the instrument one on each side by hooking their fingers under the sides using the indentations designed for the purpose Figure 2 2 A Table 3 3 Installation checklist Tick Item L Unpack the Varioskan Flash instrument carefully Refer to How to unpack on page 42 Keep the original packaging and packing material for future transportation Check the delivery for completeness Refer to Checking delivery for completeness on page 42 Check for damage during transport Refer to Checking for damage during transport on page 42 Place the instrument on a normal laboratory bench taking i
149. the contents of the reagent bottle and there is a sufficient volume of the reagent in the bottle for all priming and actual dispensing The complete dispensing tube assembly is packed with the accessories 1 Fit the complete dispensing tube assembly Figure 3 36 into the left Dispenser 1 and Dispenser 3 or right Dispenser 2 hole of the valve and tighten it finger tight The dispensing tube is used to dispense reagent from the syringe into a microplate Thermo Fisher Scientific Installation Installation setups 2 Insert the dispensing heads into their respective dispensing head holder slots on the left hand Dispenser 1 and Dispenser 3 or right hand Dispenser 2 side of the dispenser 3 Remove the protective cap which protects the sensitive dispensing tip Figure 3 35 EE ES Figure 3 35 Protective cap removed from the dispensing tip 0 40 mm Male connector of the dispensing tube assembly Aspirate tube assembly i e the input tubing incl tubing and end weight 3 port valve Plunger lock screw Complete dispensing tube assembly i e the output tubing Dispensing head Dispensing tip 2 options 0 40 mm and 0 25 mm The tubing positions depend on the flow direction of the dispenser Figure 3 36 Dispenser assembly Installation of Refer to the Skan t Software for Varioskan Flash User Manual Skanlt Software for Cat 20 N06355 for installing SkanIt Software for Varioskan Fla
150. the highest sensitivity with a limited dynamic range Medium low and Medium high ranges offer sensitivities and dynamics in between the Low and High ranges When selecting a fixed gain the principle for achieving the best sensitivity is to select the lowest possible range without receiving overrange results in the measurement The wells are measured with a selected measurement time that can vary from 10 to 1000 ms in fluorescence intensity measurements and 10 to 10 000 ms in time resolved fluorescence TRF Thermo Fisher Scientific Routine Operation Fluorometric measurement measurements There is one xenon lamp flash for each 10 ms period of measurement time The amount of xenon lamp flashes affects the quality of the measurement result Thus the more flashes the better the quality of the result The amount of flashes can be set to 1 to 100 flashes per measurement 10 1000 ms for fluorescence intensity measurements and 1 to 1000 flashes per measurement 10 10 000 ms for TRF measurements It is recommended to measure using a 100 ms measurement time in fluorescence intensity measurements and 1000 ms in TRF measurements which normally produces good results If there is a necessity to improve the quality of the results the flash amount should be increased The result is the mean value of individual 10 ms readings during the total measurement time With TRF measurements there are two additional user defined measurem
151. three dispensers with 1 ml syringe Cat no 5250510 per dispenser Thermo emorcowcnat VARIOSKAN FLASH 452 aaa Figure 1 1 Varioskan Flash spectral scanning multimode reader Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 15 Introduction to the Varioskan Flash Intended use Intended use Advantages of using Varioskan Flash 16 Thermo Scientific Varioskan Flash User Manual The Varioskan Flash spectral scanning multimode reader is intended for professional laboratory research use by trained personnel who understand the nature of fluorometry photometry and luminometry The Varioskan Flash is used to measure fluorescence intensity FI time resolved fluorescence TRF and luminescence from appropriate 6 to 1536 well plate formats or absorbance from appropriate 6 to 384 well plate formats defined by Thermo Fisher Scientific in SkanIt Software It also has incubation shaking and reagent dispensing capabilities Use for self testing is excluded Refer to Chapter 6 Technical Specifications For validation of the entire system it is recommended that Good Laboratory Practices GLP are followed to guarantee reliable analyses The Varioskan Flash provides several advantages relating mainly to the principle of operation in that it e Supports many top performance detection technologies from which to select the most appropriate for the assay in question e Supports applications requiring
152. tone MEK Methyl alcohol Methylene chloride Naphtha Nitric acid S E A S G O a G Nitric acid conc Ke Nitrobenzene Phenol Pyridine Silver nitrate Soap solutions Stearic acid Sulfuric acid Sulfuric acid conc Sulfurous acid O O o Toy Toy Toy oy rpory oy oy joy oy ToyTmoryman FC OT Oy Oy lhl Oy S AS E e oa Tannic acid Tanning extracts Tartaric acid Toluene Trichloroethylene Turpentine Water LAS r O a olojoj Xylene Thermo Fisher Scientific Thermo Scientific Varioskan Flash User Manual 83 Routine Operation Shutdown Shutd OWN To shut down the Varioskan Flash follow these steps A Warning Remove any plates or priming vessels still in the instrument Dispose of all microplates and priming vessels as biohazardous waste A 1 Rinse the dispenser tubing Figure 3 36 out thoroughly with distilled water after each use 2 If the tray is not dirty drive in the tray by selecting either Execute gt Run Plate In or clicking the oe button on the toolbar in SkanIt Software for Varioskan Flash Then switch the Varioskan Flash off by pressing the power switch Figure 2 2 on the left side panel of the instrument into the OFF position Switch the instrument off after daily use or at least if it is not in use for prolonged periods of time for example over a weekend 3 However if the
153. try Cytokine Jan 11 87 93 Kolb A Burke J and Mathis G 1997 A Homogeneous Time Resolved Fluorescence Method for Drug Discovery In Devlin J P Ed High Throughput Screening the Discovery of Bioactive Substances Marcel Dekker Inc New York Pp 345 60 Lakowicz J R 1999 Principles of Fluorescence Spectroscopy Second edition Kulwer Academic Plenum Publishers Mathis G 1999 Homogeneous time resolved fluorescence Point counterpoint J Biomolecular Screening 4 309 313 Thermo Scientific Varioskan Flash User Manual 135 References Literature Absorbance Luminescence Zaman G J R Garritsen A Boer T de and van Boeckel C A A 2003 Fluorescence Assays for High Throughput Screening of Protein Kinases Combinatorial Chemistry amp High Throughput Screening June 6 4 313 320 8 Gore Michael G Ed 2000 Spectrophotometry and Spectrofluorimetry Oxford University Press US Patent 6111636 2000 Device for measuring optical density Campbell A K Ed 1988 Chemiluminescence Principles and Applications in Biology and Medicine Horwood Chichester U K 608 pages Case J F Herring P J Robison B H Haddock S H D Kricka L J and Stanley P E Eds 2001 Bioluminescence amp Chemiluminescence World Scientific Publishing Co Pte Ltd Singapore 517 pages DeLuca M A and McElroy W D Eds 1978 Bioluminescence and chemiluminescence Methods i
154. ubing 2 Switch off the power from the instrument by pressing the ON OFF switch into the OFF position Figure 2 2 3 Loosen the plunger lock screw Figure 5 60 approximately three full turns clockwise Figure 5 61 item c 4 Pull the plunger holder arm Figure 5 60 firmly down Figure 5 61 item b 5 Unscrew the syringe from the valve Figure 5 61 item a 100 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Maintenance Periodic maintenance 6 To fit the new dispenser syringe screw the syringe into the valve pull the syringe plunger down to the plunger holder arm and screw the syringe into place Make sure the plunger lock screw is securely tightened Figure 5 61 Valve Syringe Syringe Plunger Holder Plunger Lock Screw Figure 5 61 Replacing the dispenser syringe Replacing the 3 port To replace the 3 port valve Figure 5 60 and Figure 5 62 it is valve recommended that you contact authorized technical service 1 Remove the fluid from the dispenser s 2 Remove the syringe and tubing 3 Remove the two Phillips head screws on the front of the valve then remove the valve from the dispenser s 4 Install the new valve by placing it on the front panel so the screw holes line up The valve coupler fitting mates to the valve motor shaft The valve should be oriented with the tube fittings on top and the syringe on the bottom Replace the valve screws 5 Ins
155. uch as DNA quantification with PicoGreen white plates can be used as the obtained fluorescent signal level is not very high White plates can also be used in for example GFP quantification Q4 Can all the three dispensers be used with all the detection technologies A Yes they can However depending on the dispensing position used for dispensing an extra plate movement may be carried out before the measurement step This may cause minor time delays Q5 What plates can be used with the Varioskan Flash A5 In photometry 6 to 384 well plates can be used and in fluorescence intensity time resolved fluorescence and luminescence 6 to 1536 well plates can be used Dispensing can be carried out with 6 to 384 well plates Thermo Scientific Varioskan Flash User Manual 119 Frequently Asked Questions Q amp As Table 7 16 Plates used with the Varioskan Flash Detection technology Plates recommended Fluorescence intensity Fl Black plates OR black plates with clear bottom for bottom reading Time resolved fluorometry TRP White plates OR occasionally black plates TR FRET assays OR white or black plates with clear bottom for bottom reading Absorbance Abs Clear flat bottom plates OR clear flat bottom plates with white or black walls Luminescence White plates Q6 What kind of trays and adapters are available for the Varioskan Flash A6 The trays and adapters available are listed in Table 7 17 Table
156. uots as needed The activity of the reconstituted rL L Reagent diminishes roughly 15 after 2 days of storage at 4 C Be sure to allow the rL L Reagent to return to room temperature prior to use If long term storage is needed the reconstituted rL L Reagent can be stored in single use aliquots at 20 C Avoid multiple freeze thaws The activity of the reconstituted rL L Reagent diminishes by roughly 50 after two weeks at 20 C ATP Standard Pour a small volume of ATP standard solution approximately 2 ml into a disposable 2 ml Eppendorf tube to prevent the solution from getting contaminated The ATP standard solution is ready for use The ATP Standard should be divided into 200 1000 ul aliquots and stored in the freezer Avoid multiple freeze thaw cycles with the ATP Standard Ensure that all reagents are at room temperature when preparing the test plate Pipette 15 pl of ATP Standard solution and 15 pl of rL L Reagent into well H12 of a white 384 well plate Mix the wells gently with the pipette tip This plate can be used for about 30 minutes after preparation 144 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Glossary absorbance optical density A logarithmic function of the transmission of a wavelength of light through a liquid log dimension A adapter The elevation device used to hold and raise the microplate in the tray AFP Alpha fetoprotein common label in FRET and TR F
157. ure 2 5 are located in the instrument housing Figure 2 2 under the dispenser sliding cover The dispensers consist of a pump with a valve a syringe 1 ml tubing and a dispensing head Figure 2 14 The 5 ml syringe is available on request The dispensers are designed for accurate dispensing The volume range and increment depend on the syringe size used With the 1000 pl syringe the increment is 1 pl and with the 5000 ul syringe it is 5 pl The upper limit of the range is the maximum well volume which is assured by SkanIt Software for Varioskan Flash when using multiple dispensings into the same well Thermo Scientific Varioskan Flash User Manual 29 Functional Description Dispensers The instrument supports simultaneous dispensing and reading enabling fast signal monitoring from the very start of the reaction The dispensers are located close to the measurement positions to achieve a low dead volume and minimal reagent consumption This is important when using expensive reagents Optimal design of the reagent bottle holder Figure 2 5 also helps in using all the reagent The combination of a special dispensing tip Figure 2 14 and the tip priming feature ensures that even very small volumes can be dispensed accurately To prevent dispensing in the wrong dispensing position the instrument is also equipped with dispensing position detection Male connector of the dispensing tube assembly Aspirate tube assembly i e th
158. ushed to the left of the lever opening bar Figure 3 31 Ensure that the two guide pins located on both sides of the tray holder Figure 3 31 are inserted into the tray Note Install the universal tray without the adapter being present A The contact pins where the electrical contacts of the incubator reside enable contact between the universal tray and the tray holder Figure 3 31 5 Fasten the tray to the tray holder by the screw fix by turning the Allen key supplied clockwise Figure 3 32 48 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Tray holder Tray frame Measurement chamber door How to setup the Thermo Fisher Scientific dispensers Installation Installation setups 6 Choose the detachable adapter according to the plate type you are using Always push the adapter by hand to the very bottom of the tray ensuring that it is even It may be a tight fit A clicking sound indicates successful installation Note The adapter will not go to the bottom of the tray if the positioning lever is in the way A Figure 3 32 Fastening the tray frame to the tray holder A close up of the positioning lever when the tray frame is out is shown in Figure 3 33 Lever opening bar Positioning lever M Measurement chamber door Tray frame oe A Figure 3 33 Close up of the positioning lever when the tray is out The three optional dispensers are factory
159. ve either the aspirate tube assembly that is the input tubing Figure 3 36 or the complete dispensing tube assembly that is the output tubing Figure 3 36 follow these steps 1 To remove either the dispensing tube or the aspirate tube assembly from the valve gently loosen the fittings manually Unscrew the fittings and remove the tubing Thermo Fisher Scientific Maintenance Periodic maintenance 2 It is recommended to replace the complete dispensing tube assembly always when replacement is necessary 3 To fit a new tubing insert the fitting into the valve and tighten it finger tight Replacing a When you replace a different sized dispensing tip refer to the SkanIt dispensing tip Software for Varioskan Flash User Manual Cat no N06355 If the instrument has the wrong information about the dispensing tip used it will affect the dispensing jet speed To replace a dispensing tip Figure 5 58 follow these steps 1 Remove the dispensing head tube Figure 5 58 from the brass tube holder Figure 5 58 by turning the dispensing head tube counterclockwise and the brass part closest to it clockwise Dispensing head position sensor Dispensing head tube Brass tube holder Dispensing tip 0 40 mm default gt Figure 5 58 Replacing the dispensing tip A 2 Replace the dispensing tip connected with a small piece of silicone tube in the dispensing tube Figure 5 59 Use the dispensin
160. vice Contact the Thermo Fisher Scientific technical service department for more details If the Varioskan Flash is exposed to potentially infectious chemical samples toxic or corrosive chemicals or radioactive chemicals waste management of the complete instrument must be carried out to ensure that there is no risk of contamination Warning Decontaminate the instrument before disposal Refer to Decontamination procedure on page 102 and Certificate of Decontamination on page 129 about decontamination A Follow laboratory and country specific procedures for biohazardous or radioactive waste disposal Dispose of the instrument according to the legislation stipulated by the local authorities concerning take back of electronic equipment and waste The proposals for the procedures vary by country Pollution degree 2 see Safety specifications on page 115 Method of disposal Electronic waste Contaminated waste Infectious waste Thermo Fisher Scientific Thermo Fisher Scientific Maintenance Disposal of the instrument WEEE symbol Thermo Fisher Scientific has contracted with one or more recycling disposal companies in each EU Member State European Country and this product should be disposed of or recycled through them Further information on Thermo Fisher Scientific s compliance with these Directives the recyclers in your country and information on Thermo Scientific products which may assist the detection of su
161. vity dynamic range Fluorescence intensity top reading lt 0 4 fmol fluorescein well gt 6 decades black 384 well plate Fluorescence intensity bottom reading lt 4 fmol fluorescein well gt 5 5 decades clear bottom black 384 square well plate Time resolved fluorescence top reading lt 120 amol Europium well gt 6 decades white 384 well plate top reading Measurement time Fluorescence intensity 10 1000 ms Time resolved fluorescence 10 10 000 ms Measurement speed Reads a 96 well plate in 15 s a 384 well plate in 45 s and a 1536 well plate in 135 s minimum kinetic interval time from A1 back to A1 Spectral scanning speed lt 2s well 400 500 nm 1 flash 2 nm steps Plate types 112 Thermo Scientific Varioskan Flash User Manual 6 1536 well plates top reading 6 384 well plates bottom reading Thermo Fisher Scientific Thermo Fisher Scientific Technical Specifications Performance specifications Table 6 11 Photometry Performance specifications Photometry Light source Xenon flash lamp Wavelength selection Double monochromators Detector Photodiode Wavelength range 200 1000 nm Bandwidth 5nm Wavelength setting resolution 1 nm Wavelength accuracy 2 nm Linear measurement range Absorbance resolution 0 4 Abs 96 well plate at 450 nm 2 0 3 Abs 384 well plate at 450 nm 2 Better than 0 001 Abs
162. ware For more details refer to Settle delay on page 73 Thermo Fisher Scientific Photometric spectrum scanning Luminometric measurement Thermo Fisher Scientific Routine Operation Photometric spectrum scanning Caution Do not open the measurement chamber door Figure 3 32 during measurement because this aborts the measurement A The phases of the photometric spectrum scanning measurement are the same as for the photometric measurement but with a continuous range of wavelengths See Photometric measurement on page 70 The air blank spectrum is also measured in photometric spectrum scanning measurements In luminescence intensity measurements the following actions are carried out by the instrument 1 If necessary the user installs the filter before starting the session Refer to Varioskan LumiSens on page 56 2 The tray is driven in 3 The Varioskan LumiSens filter selector position is rotated to the blocked position 4 In the signal level calibration procedure the instrument reads the internal luminescence reference signal in the blocked mode compares it to the value in the non volatile memory and sets a factor to correct the reading In long measurement procedures calibration is performed in a suitable phase without disturbing the measurement timing The default calibration interval is 7 5 minutes 5 The optics is selected by the user e Normal no filter e Filter e Monochromator
163. y Intended users How to use this user manual For more information Thermo Fisher Scientific About This User Manual This user manual has been written for the actual end user for example laboratory technician and provides information on the Thermo Scientific Varioskan Flash including the installation and operating instructions Read the manual in its entirety before operating the instrument This user manual has been designed to give you the information you need to e Review safety precautions e Install the Varioskan Flash e Use the Varioskan Flash in daily research use e Perform basic cleaning and maintenance procedures e Troubleshoot the instrument performance This user manual also describes all the features and specifications of the Varioskan Flash instrument Refer to Chapter 6 Technical Specifications In Chapter 8 Troubleshooting Guide you will find explanations of all error and warning messages and a problem solving guide The user should be familiar with the contents of Chapter 5 Maintenance For ordering information refer to Chapter 9 Ordering Information For software related issues refer to the Thermo Scientific SkanIt Software for Varioskan Flash User Manual Cat no N06355 Both the instrument and software manuals can be found in PDF format on the Thermo Scientific SkanIt Software for Varioskan Flash installation CD For the latest information on products and services
164. y hand 3 Lift up the dispenser sliding cover and open the measurement chamber door slightly Figure 3 24 4 Lift up the front cover of the instrument from both sides Figure 5 51 and remove it so that the storage site of the transport lock support and the support retaining screws marked 1 2 3 and 4 become accessible Figure 5 63 A 104 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Thermo Fisher Scientific Maintenance Decontamination procedure Figure 5 63 Transport lock released A and fastened B screws 1 4 shown 5 Remove the bottom screw marked number 3 Figure 5 63 A Loosen the bottom screw marked number 4 of the transport lock slightly by using the hexagonal screwdriver supplied Figure 5 63 A When you have loosened the screw number 4 so that the transport lock bar moves turn the transport lock bar into a vertical position towards the front of the instrument Figure 5 63 B Fit the transport lock bar into its groove in the tray holder Then remove the two top screws marked 1 and 2 of the transport lock from their storage site on the track mechanism bottom plate Figure 5 63 A and refit the two screws marked 1 and 2 into the holes designated for them using the hexagonal screwdriver supplied Figure 5 63 B At the same time replace the transport lock tag Thermo Scientific Varioskan Flash User Manual 105 Maintenance Decontamin
165. z Criteria A EN 61000 4 4 1995 A1 2001 1 kV Criteria B EN 61000 4 5 1995 A1 2001 2 kV line to ground 1 kV line to line Criteria B EN 61000 4 6 1996 A1 2001 3 Vms 150 kHz 80 MHz Criteria A EN 61000 4 8 1993 A1 2001 3 A m Criteria A EN 61000 4 11 1994 A1 2001 30 10 ms Criteria B 100 10 ms Criteria B 60 100 ms Criteria C gt 95 5 s Criteria C Thermo Scientific Varioskan Flash User Manual 117 Technical Specifications In conformity with the requirements 118 Thermo Scientific Varioskan Flash User Manual Thermo Fisher Scientific Thermo Fisher Scientific Q amp As Chapter 7 Frequently Asked Questions Q1 What is the wavelength range for the Varioskan Flash A1 In photometry the wavelength range is 200 to 1000 nm in 1 nm steps In fluorometry the wavelength range using top reading is 200 to 1000 nm for excitation light and 270 to 840 nm for emission light in 1 nm steps Q2 What plate colors can be used for fluorescence intensity A2 Best performance is generally obtained with black plates which also have the lowest background fluorescence However with some fluorochromes white plates can also be used for obtaining a slightly better sensitivity Transparent or white plates can be used but the sensitivity is often lower and the difference is dependent on the wavelengths used Q3 When can I use white plates in fluorometry A3 In certain assays s

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