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Varioskan LUX Technical Manual

1. Figure 3 21 Front cover removed The metallic transport lock support is fastened with four screws Figure 3 22 Two screws 1 and 2 are fastened to the tray holder and two screws 3 and 4 are fastened to the track mechanism bar 3 Remove the tray holder screws 1 and 2 and the transport lock tag Store the lock tag for future use 4 Gently push the track mechanism where screws 1 and 2 were fastened into the instrument 5 Remove screw 3 and loosen screw 4 Figure 3 22 Tray holder screws 1 and 2 and track mechanism bar screws 3 and 4 on the transport lock 6 Turn the transport lock into its horizontal storage position Figure 3 23 32 Varioskan LUX Thermo Scientific Installation Chapter 3 Figure 3 23 Screws 1 2 3 and 4 and the released transport lock 7 Tighten screw 4 Fasten screw 3 back into the same hole from which it was unfastened 8 Fasten screws 1 and 2 onto the track mechanism bar Keep the screws there until needed for future relocation or transportation of the instrument The transport lock is now in its storage position How to install the plate tray and adapter
2. 1 USB connector 2 Mains power supply connector 3 Connect the instrument to the PC using the USB communication cable supplied How to install Skanlt Software Refer to the Skanit Software Technical Manual for installing Skanlt Software Thermo Scientific Varioskan LUX 39 Chapter 3 Installation 40 Varioskan LUX Installing upgrade kits As the instrument is modular you can upgrade it with uninstalled features later The additional upgrades may include the LAT module including the luminescence module the AlphaScreen module and or the TRF module as well as up to two dispensers and the Integrated gas module IMPORTANT Installation of the upgrade kits must be carried out only by trained and authorized expert service personnel How to align the LAT measurement position Varioskan LUX has a function to optimize the LAT module luminescence AlphaScreen and time resolved fluorescence measurement position Note You do not have to align the measurement position if the LAT module has been factory installed The measurement position alignment should be run after the installation of a new LAT module and when the LAT base has been removed for cleaning or service The measurement position is determined according to the plate dimension information in the plate templates However the optimal position is somewhat different for each instrument because of individual mechanical variations between the u
3. Figure 5 49 Crosstalk shield 4 Turm the module over to expose the crosstalk shield 5 Carefully clean the dirt from the bottom using a lint free tissue or a cotton stick soaked in a mild detergent solution soap solution or 7096 ethanol Do not use formaldehyde IMPORTANT Try to avoid loosening the crosstalk shield but if necessary be careful to remount it into the original position without using extensive force Do not wash any electronic parts Thermo Scientific Varioskan LUX 71 Chapter5 Maintenance Figure 5 50 Removing the crosstalk shield 72 Varioskan LUX 6 Remove the crosstalk shield according to Figure 5 50 1 Unfasten the fastening screw 2 Remove the crosstalk shield 3 Remove the lens cover Soak a lint free tissue in lens cleaning solution or isopropyl alcohol Wipe over the optical lens in a straight movement from inside to out Do not use any circular movements when cleaning the optics IMPORTANT Do not touch the surfaces of filters or optical lenses with bare hands 10 11 12 13 Replace the crosstalk shield with the fastening screw 2 Replace the LAT module unit Fasten the two bottom screws 3 Attach the two connectors 2 Run the LAT measurement alignment Refer to How to align the LAT measurement position on page 40 Routine maintenance of the dispensers To obtain optimal performance and maximum useful life for the dispenser
4. Thermo Scientific Varioskan LUX Rev 1 0 Cat No N16045 jenuew jedluyre 2 Varioskan LUX Copyright 2015 Thermo Fisher Scientific Inc All rights reserved AlphaScreen AlphaLISA DELFIA and LANCE are trademarks of PerkinElmer Inc or its subsidiaries in the United States and other countries Decon is a trademark of Decon Laboratories Limited ENLITEN and Steady Glo are trademarks of Promega Corporation Microside SQ is a trademark of Global Technologies Inc Neoflon is a trademark of Daikin Industries Limited Virkon is a trademark of E I du Pont de Nemours and Company or its affiliates All other trademarks are the sole property of Thermo Fisher Scientific Inc and its subsidiaries Reproduction of the accompanying user documentation in whole or in part is prohibited Patents This product is protected by the following patent US 6144455 The Thermo Scientific TM Varioskan LUX also has an international patent pending 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 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 superse
5. The software also provides Help In an effort to produce useful and appropriate documentation we appreciate your comments on this Technical Manual to your local Thermo Fisher Scientific representative Safety and special notices Make sure you follow the precautionary statements presented in this guide The safety and other special notices appear in boxes Safety and special notices include the following CAUTION Highlights hazards to humans property or the environment Each CAUTION notice is accompanied by an appropriate CAUTION symbol IMPORTANT Highlights information necessary to prevent damage to system loss of data or invalid test results or may contain information that is critical for optimal performance of the system Note Highlights information of general interest Tip Highlights helpful information that can make a task easier Varioskan LUX 3 4 Varioskan LUX Instrument safety and guidelines for use 1 Always follow basic safety precautions when using the Varioskan LUX to reduce the risk of injury biohazardous contamination fire or electrical shock 2 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 3 Observe all CAUTION IMPORTANT Note and Tip statements as well
6. Plate types 6 384 well plates Thermo Scientific Table 6 9Luminometry Table 6 10Alpha Table 6 11 TRF Table 6 12 Integrated Gas Module Thermo Scientific Technical Specifications Chapter 6 Performance specifications Luminometry Wavelength selection Filter wheel with no filter and up to 8 optional filter positions Filter size Diameter 25 0 0 0 3 mm Thickness 3 5 7 0 mm Detector Photomultiplier tube Wavelength range 360 670 nm Sensitivity Dynamic range 7 amol ATP well white 384 well plate 7 decades Crosstalk 196 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 Wavelength selection Double monochromators Detector Photomultiplier tube Wavelength range 270 840 nm Emission bandwidth 12 nm Wavelength setting resolution 1 nm Wavelength accuracy 3nm Measurement time 10 10 000 ms Spectral scanning speed lt 2 2 s well 400 500 nm 1 flash 2 nm steps Plate types 6 384 well plates wavelength Performance specifications Alpha Light source LED Alpha measurement excitation 680 nm Emission wavelength selection Filter wheel with up to 8 optional filter position
7. 3 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 4 The instrument uses the optics setting the user has selected in the Skanlt Software measurement session Normal no filter Filter The normal mode uses the LAT module without placing any filters in the lightway The filter mode uses user defined and installed filter s 5 The instrument uses the dynamic range setting the user has selected in the Skanlt Software measurement session Figure 4 37 AutoRange Manual range for the luminescence normal and filter optics is High range Medium range Low range e Manual range for the luminescence monochromator optics is equivalent to the fluorescence measurement Refer to Figure 4 37 a Automatic dynamic range selection AutoRange defauli selects automatically the optimal reading range used It is based on signal intensity in the well and uses the lowest possible reading range to obtain best sensitivity Thermo Scientific Operations Chapter 4 Tip Do not adjust the dynamic range selection if you do not know which dynamic range to use AutoRange is almost always the optimal readi
8. First install the tray then the adapter The tray adapter combinations are individually coded for automatic identification 1 Gently pull the tray holder out and slide it to the left Figure 3 24 Figure 3 24 Tray holder 1 Guide pins 2 Screw fix hole 3 Contact pins 4 Lever opening bar 2 Install the tray into the tray holder Make sure you first push the positioning lever to the left of the lever opening bar Make sure the two guide pins located on both sides of the tray holder are inserted into the tray Thermo Scientific Varioskan LUX 33 Chapter 3 Installation Note Install the universal tray without the adapter 3 Fasten the tray to the tray holder by the screw fix by turning the key clockwise Figure 3 25 Figure 3 25 Tray and tray holder Figure 3 26 Close up of the positioning lever when the tray is out 4 Choose the detachable adapter according to the plate type you are using Refer to Plate trays and adapters on page 22 34 Varioskan LUX Thermo Scientific Installation Chapter 3 0000000 900000000000 0000000 0000000 0000000 0000000 0000000 0000000 Figure 3 27 Detachable adapter for a O 96 well plate OOOOCOCO0O 5 Place the adapter at the very bottom of the tray and make s
9. 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 R reference chip Reference chip present for validation of the fluorometric optics RFU or rfu Relative Fluorescence Fluorometric Units The arbitrary units in which fluorescence intensity is reported rpm Revolutions per minute S 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 Varioskan LUX 107 Glossary 108 Varioskan LUX T T tau Fluorescence lifetime of the TRF label transmittance The ratio of transmitted and incident light I WM 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 U USB Universal serial bus W Ww With w o Without Thermo Scientific Glossary Thermo Scientific Varioskan LUX 109 110 Varioskan LUX Thermo Scie
10. 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 Disposables e Pipette tips for pipetting 15 ul 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 ul with 15 ul volume e Thermo Scientific Finnpipette for 200 1000 ul volumes Product components e rL L Reagent 1 vial e Reconstitution Buffer 12 ml e ATP Standard 10 7M 1 vial ATP Free Water 25 ml not needed in this test Thermo Scientific Varioskan LUX 103 Appendix B Preparation of Luminescence Alignment Plate 104 Varioskan LUX Reconstitution of assay components 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 Reconsti
11. 96 well adapter for plate without lid To be used with the most common 96 well plate formats without lids Dispensing yes 3 96 well adapter for plate with lid To be used with the most common 96 well plate formats with lids no 4 384 well adapter for plate without lid To be used with the most common 384 well plate formats without lids Recommended for reading of 1536 well plates 15 mm high plates in all measurement modes and 10 mm high plates in fluorescence mode yes 384 no 1536 5 384 well adapter for plate with lid To be used with the most common 384 well plate formats with lids no 65 1536 well adapter for plate w o lid To be used with the most common 10 mm high 1536 well plate formats without lids in luminescence mode no Thermo Scientific Functional Description Chapter 2 80 6 48 well adapter for plate without lid yes To be used with the most common 6 to 48 well plate formats without lids 48 6 48 well adapter for plate with lid no To be used with the most common 6 to 48 well plate formats with lids 96 well adapter for PCR plate without yes lid To be used with 96 well PCR plates with NUNG product codes 230012 264605 or equivalent plates from other plate manufacturers included in standard deliveries Thermo Scientific Nunc M Robotic tray and adapter The robotic tray comes equipped
12. Refer to Table 2 1 and Table 2 2 The universal tray is for basic use Figure 2 15 It is compatible with all plate formats 6 to 1536 well plates Always use a plate adapter with a universal tray Refer to Universal tray with adapters on page 24 A robotic plate tray is for automated use with robots Figure 2 16 It is compatible with 96 to 1536 well plate formats Remove the plate adapter when using a plate with a lid on a robotic tray Refer to Robotic tray and adapter on page 25 Thermo Scientific Figure 2 15 Assembly picture of the universal tray Thermo Scientific Functional Description Chapter 2 Tray composition The tray composition is shown in Figure 2 15 and described below ESE LEE EEES O SS ES ELL DS RARAS os Lolo Sam Sam Enero EE eo ep AAA 1 Means of identification of tray adapter combinations For automatic identification of tray adapter combinations there are codes that refer to the coding System employed The adapters are also marked with a visual identification number and the text WITH LID or NO LID to help differentiate them 2 Tip priming vessel The tip priming vessel is an 8 well plate strip 1x8 Thermo Scientific Microtiter Solid Strip Assembly 3 Screw fix The tray is fastened to the tray holder which is part of the track mechanism by a screw fix Refer to How to install the plate tray and adapter on page 33 4 Position c
13. absorbance and luminescence 2 dispensers LOLOTD2 Varioskan LUX with fluorescence V V V V 2 dispensers L B000D2 Varioskan LUX with fluorescence top amp bottom top absorbance luminescence and TRF 2 dispensers LOLAOD2 Varioskan LUX with fluorescence top absorbance luminescence and AlphaScreen 1 dispenser LOLATD2 Varioskan LUX with fluorescence top absorbance luminescence AlphaScreen and TRF and absorbance 2 dispensers LBLOOD2 Varioskan LUX with fluorescence top amp bottom absorbance and luminescence 2 dispensers LBLOTD2 Varioskan LUX with fluorescence top amp bottom absorbance luminescence and TRF 2 dispensers lt lt lt lt gt Re ms LBLAOD2 Varioskan LUX with fluorescence 2 dispensers op amp bottom absorbance luminescence and AlphaScreen 2 dispensers VLBLATD2 Varioskan LUX with fluorescence top amp bottom absorbance luminescence AlphaScreen and TRF Gas module for Varioskan LUX Cat no Instrument System VLGAS001 Integrated gas module factory installed at instrument purchase Upgrade kits for Varioskan LUX Cat no Instrument System VLUPLOOO2 Luminometric upgrade kit L module service upgradable VLUPADO3 AlphaScreen upgrade kit service upgradable requires L module VLUPTOO4 TRF upgrade kit service upgradable requires L module VLUPDOO5
14. 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 us 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 fluorometry The measurement of fluorescence Also known as fluorimetry FRET Fluorescence resonance energy transfer 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 I initialization tests So called self tests which are carried out before operation to ascertain that the necessary instrument adjustments have been carried out L LANCE PerkinElmer s TR FRET kit family using Europium ACP 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 Thermo Scientific Thermo Scientific luminometer An instrument used for measuring the intensity of luminescent radiation luminometric label luminophore A
15. level measurements are required after installation IMPORTANT Do not operate the instrument in an environment where potentially damaging liquids or gases are present Technical prerequisites Place the instrument on a normal sturdy laboratory bench The net weight of the unit is 53 to 60 kg 117 131 Ibs depending on the configuration The instrument operates at voltages of 100 240 Vac and the frequency range 50 60 Hz Thermo Scientific Installation Chapter 3 Installation setups This section describes the installation setups that have to be carried out before instrument operation How to release the transport locks There is a transport lock for the tray holder in the instrument Figure 3 19 Make sure the transport lock has been released before you put the instrument into operation w NY Figure 3 19 Transport locks and transport lock tags present ST Tray holder transport lock To release the tray holder transport lock 1 Lift up the dispenser sliding cover 1 Figure 3 20 Figure 3 20 Dispenser sliding cover opened Thermo Scientific Varioskan LUX 31 Chapter 3 Installation 2 Remove the front cover by pulling the two bolts 1 one on each side at the bottom sideways then turning the bottom of the cover 2 30 and lifting it off according to Figure 3 21
16. ow Benzyl alcohol Boric acid Bromine ojo Butyl alcohol k Butyl acetate Carbon sulfide Carbon tetrachloride Chloroacetic acid Chlorine ojpjojojojojojojojo o jo o Chlorobenzene Chloroform Chromic acid Cresol Cyclohexane Dimethyl sulfoxide DMSO oj Ethers Ethyl acetate Ethyl alcohol Ethyl chromide Formaldehyde Formic acid Freon Gasoline Glycerin Hydrochloric acid Hydrochloric acid conc ojojoilco o o co jco o jc Hydrofluoric acid Hydrogen peroxide Hydrogen peroxide conc Hydrogen sulfide Kerosene CeOIiojojojo o joj ojo nio o ji ethyl ethyl ketone MEK ethyl alcohol ethylene chloride aphtha tric acid tric acid conc ojoju o ojo jojoj o 2 itrobenzene Phenol Pyridine Silver nitrate ojojojojojojojojo oj joj jojojojojojojo o jo jojo o o o o oO Jo Jo jo Oo Continued Varioskan LUX 63 Chapter 4 Operations 64 Varioskan LUX Cont Solvent Soap solutions Stearic acid Sulfuric acid Sulfuric acid conc Sulfurous acid ojojojojoljo Tannic acid Tanning extracts Tartaric acid Toluene Trichloroethylene Turpentine Water Xylene ojjojojo
17. Checking for damage during transport Visually inspect the transport package the instrument and the accessories for any possible transport damage If the box has 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 If any parts are damaged contact your local Thermo Fisher Scientific representative or Thermo Fisher Scientific Oy Environmental requirements When you install Varioskan LUX avoid sites of operation with excess dust vibrations strong magnetic fields direct sunlight draft excessive moisture or large temperature fluctuations Make sure that e The working area is flat dry clean and vibration proof and leave additional room for cables covers and so on e There is at least 10 cm of free space around the instrument on the laboratory bench for ventilation e There is sufficient room behind the instrument to enable disconnecting the device e The ambient air is clean and free of corrosive vapors smoke and dust e The ambient temperature range is between 10 C 50 F and 40 C 104 F e Humidity is low so that condensation does not occur relative humidity is between 10 and 80 non condensing e CO and O sensors are present on the wall of the room where the Varioskan LUX with a Integrated gas module is installed The Varioskan LUX does not produce operating noise at a level that would be harmful No sound
18. DR le a 22 Plate trays and Adapters i a atre rtm etm et estie eb maa p eet E 22 Tay COMPOSION iva eros eo ep taa 23 Universal tray with adapters 0 H He 24 Robotlc fray and adapter teet et eren essct CY aee Y a t us 25 How to change the location of the holder for the tip priming vessel 26 How to remove or replace the robotic tray adapter 0oooo ooooo momo 21 Ghapter 3 Installation siii a a a A ee 29 What to do upon delivery aaiae neo ia aga enter eee Hee 29 POW 10 UNDACK os erede Ce ae AA tea e Y DI si De d 29 Checking delivery for completeness ooo cocina RERRRRI vet es 30 Checking for damage during tansport s scii mii eaa e 30 Environmental requirements o oo oooooooocccocor Hn 30 Technical pre QUISTES ee ree dt e Re rm ee E ns 30 Installation SetuDS iiie Hm Hm RR 31 How to release the transport locks isssssssse ee eet teens 31 Tray holder transport loCcK 60 e R IIR 31 How to install the plate tray and adapter n n ccc eee teen ens 33 How to Set up ThE disperiSelis porra Rp ekz EeEPXerYessaweg meer 35 How to set up the Integrated gas Module 6 e 37 Thermo Scientific Varioskan LUX 5 Table of Contents 6 Varioskan LUX HOW TO CONNECE Ci ia eee e Rea N 39 How to install Skanlt Software 0 Hn 39 stalling Upgrade Sia as 40 How to align the LAT measurement position oooooooooorooroooomo o 40 How to install LAT module fil
19. Discovery Edition Filters for Varioskan LUX Table 8 25 Filters Contact your local Varioskan LUX sales representative for the latest list of available TRE AlphaScreen AlphaLISA and luminometric filters Thermo Scientific Varioskan LUX 99 Certificate of Decontamination Name Address Tel Fax Instrument Serial No A confirm that the returned items have not been contaminated by body fluids toxic carcinogenic or radioactive materials or any other hazardous materials B 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 letters Please include decontaminating solution used Thermo Scientific Varioskan LUX 101 Preparation of Luminescence Alignment Plate Required reagents materials and equipment These required reagents materials and equipment are not provided by Thermo Fisher Scientific unless otherwise indicated 1 Reagents
20. Dispenser upgrade kit service upgradable VLUPGOO6 Integrated gas module upgrade kit Service upgradable Accessories for Varioskan LUX Cat no Instrument System N12391 uDrop Plate NO3079 Robotic Tray including adapter for plate without lid N02692 96 well adapter for plate without lid N02693 96 well adapter for plate with lid N02690 384 well adapter for plate without lid N02691 384 well adapter for plate with lid N02696 6 48 well adapter for plate without lid N02697 6 48 well adapter for plate with lid N06210 1536 well adapter for plate without lid N03395 96 well adapter for PCR plate without lid N02339 Adapter for plate without lid Continued Thermo Scientific Ordering Information Chapter 8 Cont N16443 Adapter holder N16484 Filter pick up tool 2805690 Aspirate tube assembly for dispenser SP 00094 Dispensing tube assembly SP 00096 Dispenser syringe 1 ml SP 00093 Dispenser syringe 5 ml N164263 Varioskan LUX Demo instrument box Verification tools for Varioskan LUX Table 8 23 Catalog numbers for m Instrument System verification tools N03394 Spectrophotometric verification plate 2806460 Lumiwell Verification Plate Skanlt Software Table 8 24 Catalog numbers for Skanlt Cat no Instrument System Software 5187139 Skanlt SW for Microplate Readers Research Edition 5187149 Skanit SW for Microplate Readers Drug
21. IMPORTANT If the dispensers are not properly installed leakage may occur Thermo Scientific Figure 5 51 Replacing the dispensing tip A Thermo Scientific Maintenance Chapter 5 Replacing the aspirate tube assembly or the complete dispensing tube assembly To remove either the aspirate tube assembly that is the input tubing Figure 3 30 or the complete dispensing tube assembly that is the output tubing Figure 3 30 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 2 Insert the fitting into the valve and tighten it finger tight to fit the new tubing Replacing a dispensing tip The dispensing tip replacement procedure is carried out in two stages e Manual replacement of the dispensing tip e Defining the dispensing tip data using Skanlt Software if you have changed the tip size If the instrument has the wrong information about the dispensing tip size used it will affect the dispensing jet speed Refer to the Skanit Software Technical Manual To replace a dispensing tip Figure 5 51 1 Remove the dispensing head tube Figure 5 51 from the brass tube holder Figure 5 51 by turning the dispensing head tube counterclockwise and the brass part closest to it clockwise Dispensing tip 0 40 mm default Dispensing head tube 3 Dispensing head position sensor Brass tube holde
22. Refer to How to set up the dispensers on page 35 Dispensing This section provides valuable information on how to use and maintain the optional dispensers IMPORTANT When using a dispenser make sure the aspiration tube end is completely submerged in the contents of the reagent bottle and there is a sufficient volume of the reagent in the bottle for all priming and actual dispensing IMPORTANT The instrument carries out automatic checks but nevertheless make sure you do not dispense into the instrument by mistake Ensure that e A correct microplate has been inserted into the plate tray e The microplate or tip priming vessel is not too full The recommended Check plate before session execution in Skanlt Software Settings gt General settings is ticked Thermo Scientific Varioskan LUX 57 Chapter 4 Operations Figure 4 44 Identification of the dispensing tip position Table 4 4 Readings carried out in measurement positions F and L 58 Varioskan LUX O D d j F O O Ox The two dispensing positions are marked F and L Figure 4 44 The F and L dispensing positions can be used with either Dispenser 1 or 2 Dispensing position F points the dispensing head towards the measurement position F Dispensing position L points the dispensing heads towards the measurement position L The readings that are carried out in both of these measurement positions are presented in Table 4 4 If you want to
23. The user loads the cell plate onto the device and starts the kinetic run in which the desired reaction is followed 4 The session can maintain measurements using any measurement technology as well as shaking dispensing and incubating adjustments The system contains warning mechanisms in case of any error situations occurring Other functions The Varioskan LUX also has shaking incubating and reagent dispensing features which are presented below Track mechanism The track mechanism Figure 4 39 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 start up 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 Varioskan LUX 53 Chapter 4 Operations Figure 4 39 Part of the Varioskan LUX track mechanism Figure 4 40 ON OFF period time and total shaking time 54 Varioskan LUX 1 Tray holder 2 X carriage 3 Tray 4 Y carriage Orbital shaking The orbital shaking function is used for shaking the microplate to mix the samples Movement of the track mechanism Figure 4 39 can perform the shaking action
24. V c i ste bp e LR LELREYA DEPO qp tot eee Sie Pon nad aper saeua 105 Varioskan LUX 7 Figure 1 1 Thermo Scientific TM Varioskan LUX Thermo Scientific Introduction to Varioskan LUX Overview Thermo Scientific Varioskan LUX Figure 1 1 is a modular multi technology microplate reader Varioskan LUX is controlled by Thermo Scientific Skanlt Software for Microplate Readers End point kinetic and spectral measurements can be carried out in the UV Vis NIR range from appropriate microplate formats In fluorescence intensity time resolved fluorescence luminescence and AlphaScreen measurements 6 to 1536 well plates can be used and correspondingly 6 to 384 well plates in absorbance measurements The instrument is always equipped with the following detection technologies e Absorbance e Fluorescence intensity Fl Depending on the instrument model the following optional detection technologies LAT module may be included e Luminescence e AlphaScreen e Time resolved fluorescence TRF Varioskan LUX 9 Chapter 1 Introduction to Varioskan LUX The instrument selects the measurement wavelength either by using filters or monochromators depending on the measurement technology Monochromators are used in absorbance and fluorescence intensity measurements Filters are used in AlphaScreen and TRF measurements Most luminescence measurements do not require any wavelength selection But if
25. 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 Absorbance Each selected wavelength is calibrated 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 expiry period is one hour in absorbance Recalibration is performed 45 minutes after the previous calibration depending on the measurement procedure Fluorescence intensity amp Time resolved fluorescence The automatic calibration expiry period is 10 minutes Recalibration is performed 7 5 minutes after the previous calibration depending on the measurement procedure Luminescence amp AlphaScreen The automatic calibration expiry period is 10 minutes Recalibration is performed 7 5 minutes after the previous calibration depending on the measurement procedure In luminescence calibration a dark level control measurement is also performed to compensate for possible electronic component and photomultiplier signal level drift Measurements and scanning CAUTION Do not open the measurement chamber door Figure 3 25 durin
26. and do not let any cleaning fluids enter the measurement chamber Figure 5 47 Use external containers Weak detergent or 1096 bleach To clean the dispensers Figure 3 28 with weak detergent or 10 bleach 1 Prime the dispenser with a weak detergent solution or a 1096 bleach solution Make a solution of 1096 bleach by adding one part of commercial bleach to nine parts of water Leave the solution in the dispenser with the syringe Figure 3 30 fully lowered for 30 minutes 2 After the 30 minute period remove the aspirate tubing Figure 3 30 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 Weak base and acid in sequence To clean the dispensers Figure 3 28 with weak base and acid 1 Prime the dispenser with 0 1 M NaOH and leave the solution in the dispenser for 10 minutes with the syringe Figure 3 30 fully lowered Varioskan LUX 73 Chapter5 Maintenance 74 Varioskan LUX IMPORTANT Do not spill any alkalines onto any instrument surfaces to avoid damage to the instrument If needed use suitable protection covering 2 Flush the dispenser with distilled or deionized water 3 Prime the dispenser with 0 1 M HCI and leave the solution in the dispenser for 10 minutes with the syringe fully lowered 4 After the 10 min
27. are aligned 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 Install the syringe and pull the syringe plunger until it is above the carriage Align the valve using the plunger as a guide and tighten from 1 8 to turn after the syringe touch off Pull the syringe plunger all the way into the carriage and secure by tightening the plunger lock screw Thermo Scientific Maintenance Chapter 5 Valve Tubing Ports E T 9 Valve Plug Valve o gt e y Screw Mounts Valve Coupler Fitting Figure 5 55 3 port valve replacement Front View Syringe Mount Rear View Not shown Always located at bottom Disposal information Disposal of materials Follow laboratory and country specific procedures for biohazardous or radioactive waste disposal Refer to local regulations for the disposal of infectious material CAUTION The samples can be potentially infectious Dispose of all used plates strips priming vessels disposable gloves syringes disposable tips and so on as biohazardous waste Disposal of the instrument If the Varioskan LUX 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 CAUTION Decontaminate the inst
28. concentration below target Target CO concentration below target 99 CO sensor failure CO sensor failure Thermo Scientific Table 7 17 Warning codes reported Code 100 NEEL Unable to comply with the defined lag time Troubleshooting Chapter 7 Suggested action 101 Unable to comply with the defined well interval 102 103 Unable to comply with the defined wavelength interval Unable to comply with the defined kinetic interval 104 Unable to comply with the defined group interval 105 The timer referenced in the WAI timer command is not anymore running Your timing requirement is not met Make sure you started the timer with a long enough wait time 106 The lamp lifetime has reached its end Arrange for the replacement of the lamp as soon as convenient 107 Calibration validity has expired The accuracy of the measurement results may have suffered The action depends on the calibration options your PC offers and the type of assay you were running If there is no waiting time in the assay then you have to accept the possible accuracy reduction If there is waiting time you could switch on the automatic calibration feature if that is an option in the PC software 108 Command has no effect This just informs that a command has been used which has no effect in the current measurement method Commands BWS PAR 4 PAR 5 PAR 8 PMT and SET 9 are no o
29. heads properly into the dispensing head positioners IMPORTANT Carefully insert the dispensing head straight into the dispensing head holder without damaging or contaminating the dispensing tip Thermo Scientific Varioskan LUX 59 Chapter 4 Operations 60 Varioskan LUX Tip priming The instrument has a Prime tip ul feature see the Skanlt Software Technical Manual This function can be selected in Skanlt Software or it can be turned off If selected the dispenser automatically dispenses 2 to 10 ul reagent with the 1 ml syringe and 5 or 10 ul 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 within the range of 2 to 50 ul 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 Note Tip priming is a different procedure from manual priming that must be performed when a reagent bottle Figure 3 28 is installed next to the dispenser and the dispenser tubes are completely empty The tip priming vessel is an 8 well plate strip 1x8 Thermo Scientific Microtiter M Solid Strip Assembly There is a cavity for the tip priming vessel in the left side of the universal tray position A Figure 2 15 The robotic tray has two optional locations for
30. in appropriate microplates The instrument also has incubating atmospheric control shaking and reagent dispensing capabilities It is used with an external computer control software The reader is intended to be used in research laboratories by professional personnel The multi technology reader is not intended for diagnostic use For validation of the entire system it is recommended that Good Laboratory Practice GLP is followed to ensure reliable analyses Varioskan LUX 11 Functional Description Instrument layout Front view The front view of the Varioskan LUX instrument is shown in Figure 2 2 MU Power switch LED indicator Dispenser sliding cover Front cover Measurement chamber door A Pon Figure 2 2Varioskan LUX front view Thermo Scientific Varioskan LUX 13 Chapter 2 Functional Description Back view The back view of the Varioskan LUX instrument is shown in Figure 2 3 and Figure 2 4 PO 4 Z OF Figure 2 3 Varioskan LUX back view meee e gt Cooling fan outlets USB connector Mains power supply connector Power switch Gas connectors opi Or S oN Figure 2 4 Close up of the computer and mains supply connectors 1 USB connector 2 Mains power supply connector 14 Varios
31. is only Contact service reported in response to the FCA command 40 Emission gratings 405 nm calibration failed This is only Contact service reported in response to the FCA command 41 Emission gratings 695 nm calibration failed This is only Contact service reported in response to the FCA command 42 Flash lamp calibration failed This is only reported in Contact service response to the FCA command Continued Thermo Scientific Thermo Scientific Troubleshooting Chapter 7 Cont Code Explanation Suggested action 43 PMT relative spectral sensitivity calibration failed This is Contact service only reported in response to the FCA command 44 PMT gain calibration failed This is only reported in Contact service response to the FCA command 45 XY table position calibration failed This may be reported Contact service after start up or in response to the FCA command 46 No factory calibration for the current measurement After adding hardware for a new measurement method method the method cannot be used until the instrument is recalibrated by giving the FCA command 47 Plate tray alignment error This is only reported in Check that the plate tray is properly inserted response to the FCA command Contact service 48 LAT module calibration failed This is only reported in response to the FCA command 49 No plate tray attached to the X carriage The XY table will not move unless there is a plate tr
32. nominal Power consumption 200 VA max User interface The instrument is under PC software control Computer interface USB 2 0 Measurement types Fluorescence intensity time resolved fluorescence photometry luminometry and AlphaScreen Incubator Incubator included heating Shaker Orbital shaking Dispensers Up to two 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 LUX 85 Chapter 6 Technical Specifications Table 6 7 Fluorometry Table 6 8 Photometry 86 Varioskan LUX Performance specifications in room temperature This section provides the performance specifications for the relevant measurement techniques and other instrument capabilities 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 12nm Wavelength setting resolution 1nm Wavelength accuracy Excitation 2 nm emission 3 nm Xenon flash lamp Lamp lifetime typically 10 flashes 109 96 well microplates using 100 ms integration time per well Sensitivity dynamic range Top reading 0 4 fmol f
33. of Luminescence Alignment Plate Refer to the Skanit Software Technical Manual on how to start the measurement position alignment in Skanlt Software How to install LAT module filters The LAT module has built in AlphaScreen and TRF excitation filters You need to install the AlphaScreen TRF and luminescence emission filters IMPORTANT Do not touch the surfaces of filters with bare hands 1 Turn on the instrument and open Skanlt Software 2 Open the dispenser sliding cover and the LAT module cover 3 Select the filter position in Skanlt Software a Click Settings on the application menu b Click Instruments c Click the icon on the right side of the instrument name to open the Edit instrument parameters window d Click the Filter definition tab e Click Add f Selecta free filter position from the filter wheel and add the new filter information g Click Next The filter wheel is now turned to the selected position General Dispensers LAT module Report Filter definition Figure 3 34 The Add new filter pop up window Net gt Cancel 4 Open the blue filter nest lid on the LAT module 5 Loosen the filter wheel screw on the selected position 6 Place the filter on a clean dust free surface with the arrow on the side of the filter pointing upwards 7 Use the filter pick up tool to place the filter into the bottom of the filter nest Figure 3 35 8 Tighten the filter wheel
34. page 35 When the dispensers are not in use the dispensing heads can be stored in the dispensing head holders Figure 2 5 IMPORTANT To avoid light from entering the measurement chamber use blind plugs to close empty dispensing positions Priming Priming means filling the tubing with liquid to be dispensed The dispensers can be primed using Skanlt Software or control buttons for priming the dispensers Prime the dispenser tubing if necessary Either click on the Prime icon in Skanlt Software or use the Prime control button located on top of each dispenser Figure 2 5 and Figure 2 14 1 To prime remove the dispensing heads Figure 2 5 from 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 2 Insert an empty plate of the same type as the actual assay plate into the tray For safety reasons there must be a plate on the tray during priming 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 incorrectly inserted into the dispensing head positioners during priming 3 Press the Prime button and visually check that the dispensing jets are straight 4 Insert the dispensing
35. screw Thermo Scientific Varioskan LUX 41 Chapter 3 Installation OS Figure 3 35 Filter nest and pick up tool CS P 1 Filter nest 9 Close and fasten the filter nest lid 10 Click Finish 42 Varioskan LUX Thermo Scientific Thermo Scientific Operations The operation of the Varioskan LUX modular multi technology reader is controlled by an external computer and run on Skanlt Software CAUTION Do not operate your instrument from a power outlet that has no ground connection CAUTION Do not smoke eat or drink while using the Varioskan LUX Wash your hands thoroughly after handling test fluids Observe normal laboratory procedures for handling potentially dangerous samples Use proper protective clothing Use disposable gloves Ensure that the working area is well ventilated Do not spill fluids in or on the equipment IMPORTANT Operate the instrument only with software and hardware specifically designed for it Thermo Fisher Scientific assumes no liability for the use of third party software applications IMPORTANT It is recommended that the assay includes internal quality control samples to verify operation Operational check First switch the Varioskan LUX 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 a
36. the light source and the wavelength selection devices CAUTION Do not open the optical covers under any circumstances There is a risk of ultraviolet radiation injury Only authorized service personnel have permission to open the optical covers Thermo Scientific Varioskan LUX 17 Chapter2 Functional Description Figure 2 7 Excitation optics Figure 2 8Principle of the double monochromator 18 Varioskan LUX Diffraction Xenon order 1 Mono 2 Mono Bandwidth flash selection chromator chromator selector Excitation light module filters Light source A xenon flash lamp is used as the light source The lamp provides the wide spectral range needed for absorbance and fluorescence 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 measurement quality and measurement speed requirements Diffraction order selection filters Excitation diffraction order filters namely 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
37. 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 16 Tip priming vessel in holder position A portrait orientation Positioning lever Figure 2 16 Robotic tray fitted with adapter for plate w o lid 126 Robotic tray frame Remove the adapter when you use microplates with lids Refer to How to remove or replace the robotic tray adapter on page 27 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 Refer to Table 2 2 and Chapter 8 Ordering Information Thermo Scientific Varioskan LUX 25 Chapter2 Functional Description Table 2 2 Compatibility of the robotic tray and plate specific adapter Holder for tip priming vessel position A portrait orientation position B landscape orientation Figure 2 17 Changing the location of the tip priming vessel holder Washer and retaining screw for holder for tip priming vessel 26 Varioskan LUX Picture ID no Description Dispensing 126 Robotic tray with adapter for plate yes 96 amp 384 without lid no 1536 To be used with the most common 96 to 1536 well plate formats without lids 127 Robotic tray without adapter for plate no with lid To be used with the most common 96 to 1536 well plate formats with lids
38. 5 Chapter 7 Troubleshooting Table 7 18 Troubleshooting guide 96 Varioskan LUX Troubleshooting guide The problems covered below are considered faults that require repair or corrective work Table 7 18 If problems occur or reoccur contact authorized technical service immediately Symptom Connection to the instrument fails Cause The instrument is switched off The instrument is not completely initialized Suggested action Switch on the instrument Wait for the initialization tests to be performed An empty well or an incorrect layout vs actual layout Measurement at wrong wavelength The USB cable is not connected Connect the USB cable The USB cable is broken Replace the USB cable Too low signal or no signal at all The plate is missing Insert the plate Check that the plate vs layout matches Measure with the correct wavelengths Poor measurement precision and Incorrect plate template Select Modify the plate template to inconsistent results correspond to the used microplate Inconsistent or bad Incorrect or missing adapter with or Place the correct adapter onto the measurement results without lid plate tray Too high results in fluorescence Excitation and emission wavelengths chosen too close to each other Excitation and emission wavelengths have to be correct in relation to the used bandwidth Inconsistent results in LAT measurements Either t
39. Figure 2 10 is basically similar to excitation optics Refer to Excitation optics on page 17 CAUTION Do not open the optical covers under any circumstances There is a risk of ultraviolet radiation injury Only authorized service personnel have permission to open the optical covers Diffraction Photo Emission light 3 Mono 4 Mono order multiplier from the well chromator chromator selection tube UT UT LL filters PMT Emission monochromators Two diffraction grating monochromators are connected serially as in excitation optics to gain high stray light rejection The monochromator bandwidth is 12 nm Emission diffraction order filters Emission diffraction order filters are used to block unwanted harmonic transmission from the monochromators Varioskan LUX 19 Chapter2 Functional Description Figure 2 11 Absorbance measurement module Figure 2 12 Fluorescence bottom reading optics 20 Varioskan LUX 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 Besides fluorescence the emission optics is also used for generating luminescence spectra Absorbance measurement module Absorbance measurement Figure 2 11 is carried out by using the excitation optics module as the absorbance measurement light source Reference sensor Meas
40. How to change the location of the holder for the tip priming vessel 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 17 or in the front position B Figure 2 17 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 andscape orientation or vice versa Figure 2 1 7 1 Remove the holder for the tip priming vessel Figure 2 17 by unfastening the holder retaining screw Figure 2 1 7 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 17 The guide pin controls that the holder is placed correctly Then fasten the holder retaining screw fitted with a washer Position A A Guide pins Thermo Scientific Functional Description Chapter 2 How to remove or replace the robotic tray adapter The elevation adapter is used for microplates without lids Remove t
41. Principle of the optical system LAT module LAT Emission Spectral scanning module Excitation optics Dispensing position L Y Measurement position L Dispensing position F Y Photometer beam through the plate Photometer Fluorometer optics bottom optics Emission fiber Emission Excitation Excitation fiber Thermo Scientific Functional Description Chapter 2 The Varioskan LUX optical unit consists of six subunits Figure 2 6 e Excitation optics produces light of selected wavelength for fluorescence measurement and also for absorbance measurement The excitation optics module consists of the light source and wavelength selection devices namely diffraction order selection filters and monochromators Refer to Excitation optics on page 17 Measurement optics produces a high definition optical beam for fluorescence and absorbance measurements at measurement position F Figure 4 44 and Table 4 4 The excitation light reference detector is incorporated into the measurement optics module Simultaneously the measurement optics module collects emission light which is fed to the emission reading channel Refer to Measurement optics on page 19 Emission optics carries out the reading of a selected wavelength for fluorescence and spectral scanning module based luminescence The emission optics module is basically similar to the excitation optics modu
42. The shaking action can be intermittent and consist of so called ON shaking periods and OFF times pause periods between shaking periods Figure 4 40 or be a constant shake You can also select whether the shaking sequence starts or ends with a shaking ON time Refer to the Skanlt Software Technical Manual The speed and shaking force can be selected by the user in the Skanlt Software session The speed is adjustable from 60 to 1200 rpm revolutions per minute in 60 rpm increments The shaking force selection is Low Medium or High and corresponds to a certain diameter of the orbital movement Centrifugal forces greater than 1 G are automatically prevented IMPORTANT The system only allows certain shaking speed and force combinations to help avoid liquid spillage inside the instrument However do not fill the wells too full Figure 5 47 Thermo Scientific Upper heater slightly tilted Top reading window Figure 4 41 Varioskan LUX incubator cross section Thermo Scientific Operations Chapter 4 Incubator The instrument has an accurate incubator for temperature control up to 45 C It is useful for temperature critical applications for example certain enzyme assays and cell based applications Set the temperature via Skanlt Software The universal tray is specially designed for precise and uniform temperature control The incubator consists of two main parts Figure 4 41 A fixed upper heater and A movi
43. UAD analog board gain steps failed Contact service 25 Analog signal outside measuring range This situation is Contact service an error during start up and the execution of the CAL command 26 Flash lamp failure Contact service 27 Attempt to set the instrument serial number when it Do not try to set the serial number already has been set 29 A dispenser is not responding Contact service 30 Non volatile parameters lost Contact service 31 The digital to analog converters check of the QUAD Contact service analog board failed 32 The requested measurement method is not available Do not try to use measurement methods not supported by the instrument 33 Dispensing or priming was attempted when there was no Insert a plate into the plate tray plate inserted into the plate tray 34 Dispenser tip priming was attempted when there were Insert the waste wells no waste wells inserted into the plate tray 35 Excitation gratings zero reflection angle not found This is Contact service only reported in response to the FCA command 36 Excitation grating 1 335 nm calibration failed This is only Contact service reported in response to the FCA command 37 Excitation grating 1 823 nm calibration failed This is only Contact service reported in response to the FCA command 38 Excitation grating 20 nm calibration failed This is only Contact service reported in response to the FCA command 39 Excitation grating 2 823 nm calibration failed This
44. alibration hole s There are three holes for the automatic calibration and alignment of the track mechanism The hole marked is used for the positional check of the track mechanism at start up and when changing the tray 5 Holder for tip priming vessel The plate tray has a cavity for the tip priming vessel 6 Adapter The adapter is a detachable part that is fitted into the universal tray It lifts the microplate to the optimum height for measurement and dispensing 7 Positioning lever The positioning lever is used for automatic positioning of the microplate 8 Universal frame Varioskan LUX 23 Chapter2 Functional Description Table 2 1 Compatibility of the universal tray and plate specific adapters 24 Varioskan LUX Universal tray with adapters Plate adapters are used for adapting plates that are of different heights Adapters lift the plates to the optimum height for measuring and dispensing Before you run a measurement check that the correct plate adapter is in the plate tray Choose the adapter based on the plate format you have and whether you are using a lid or not IMPORTANT Do not use plates with dimensions exceeding the top rim of the tray Note that the maximum total height of plates is manufacturer specific Refer to Table 2 1 and Chapter 8 Ordering Information For more information on plate type settings refer to the Skanit Software Technical Manual ID no 2 Adapter Description
45. annot be changed Thermo Scientific Varioskan LUX 37 Chapter 3 Installation Figure 4 31 Supplied tubes for O left and CO right 1 Attach the gas supply system to the filters using silicon tubing or similar with 4 mm internal diameter 2 Attach the connectors at the other ends of the tubes to the gas connectors on the back panel of the instrument see Figure 4 32 and Figure 2 3 Attach CO gas line to the connector 1 and N gas line to the connector 2 3 p Figure 4 32 Gas connectors p 7 1 Connector for CO gas line 2 Connector for N gas line 38 Varioskan LUX Thermo Scientific Installation Chapter 3 How to connect cables This section shows the location of all relevant connectors and how to connect the mains supply cable CAUTION Ensure that the mains switch Figure 2 2 on the left side panel is in the off position Do not operate your instrument from a power outlet that has no ground connection 1 Connect the mains supply cable to the mains power supply connector Figure 3 33 on the back 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 om o Figure 3 33 Connecting the mains y supply cable
46. as safety symbols and markings on the instrument and in the documentation 4 Do not open any other covers of the Varioskan LUX than the dispenser sliding cover Figure 2 2 or measurement chamber door Figure 2 2 while the instrument is plugged into a power source 5 Do not open the measurement chamber door while the instrument is in operation when the LED indicator is orange 6 Do not push in the tray manually unless the instrument is switched off 7 Do not force a microplate into the instrument 8 The Varioskan LUX 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 Practice GLP is followed to ensure reliable analyses 9 Follow the preventive maintenance instructions closely to keep the instrument in the best condition for maximum reliability A poorly maintained instrument will not give the best results Emergency situations If there is any abnormal situation during the operation such as fluids spilling inside the instrument 1 Switch off the instrument Figure 2 2 2 Unplug the instrument immediately from the power supply Figure 3 33 3 Carry out appropriate corrective measures However do not disassemble the instrument If these corrective measures taken do not help contact authorized technical service or your local Thermo Fisher Scien
47. ay 50 Too high background level The background level is checked during execution of the CAL command Contact service Attach a plate tray to the X carriage of the XY table Clean away any possible liquid spills inside the measurement chamber Contact service if the error persists 51 A dispenser has failed to initialize properly Contact service 52 A 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 Valve position does not allow moving the plunger Contact service 65 Dispenser command buffer overflow Contact service 66 Attempt to dispense when the dispenser is not primed Prime the dispenser before dispensing 67 Dispenser is not installed Do not try to use nonexistent dispensers 68 The drift compensation factor is too far from the nominal Contact service value of 1 0 69 Not enough memory for a new user defined parameter Using the PUT command delete some unused This error may be reported in response to a PUT use
48. 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 namely stray light 2 4 rotating concave grating y d Faint stray light Faint stray light diffraction ia diffraction Input slit y Intermediate slit Exit slit Diffracted light amp faint stray light Selected pure wavelength Racal 1 rotating concave grating Thermo Scientific Figure 2 9 Measurement optics Figure 2 10 Emission optics Thermo Scientific Functional Description Chapter 2 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 for fluorescence measurement and for absorbance measurement Simultaneously the measurement optics collects emission light which is fed to the emission reading channel Reference sensor Excitation light Emission light 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 Emission reading module Emission optics
49. ctive glasses and clothing Decontamination should be performed in accordance with normal laboratory procedures Any decontamination instructions provided with the reagents used should be followed It is strongly recommended to perform the complete decontamination procedure before relocating the instrument from one laboratory to another Example of decontaminants Ethanol 7096 e Virkon solution 1 3 Glutaraldehyde solution 4 Chloramine T Microcide SQ 1 64 Decon 90 min 4 IMPORTANT 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 inconsistent test results Thermo Scientific Thermo Scientific Maintenance Chapter 5 To decontaminate the instrument 1 Prepare the decontaminant 200 ml 4 glutaraldehyde solution or another agent recommended by your safety officer 2 Empty the tray Figure 2 15 and Figure 2 16 Ensure that you are wearing disposable gloves 3 Switch off the power Figure 2 2 and disconnect the mains supply cable Figure 3 33 4 Disinfect the outside of the instrument using a cloth dampened with 70 ethanol 5 Place the instrument in a large plastic bag Ensure that the dispenser sliding cover and front cover Figure 3 21 are open and the tray is out Figure 4 36 6 Place a cloth soaked in the prepar
50. cts the signal during the defined TRF integration time These actions form one TRF measurement cycle Figure 4 38 which is performed within a 10 ms period The cycle is repeated as many times as defined by the measurement time Thermo Scientific Figure 4 38 Structure of the TRF measurement cycle Thermo Scientific Operations Chapter 4 RFU Background fluorescence Y A Y Time us Lamp Delay Measurement flash time time 6 lf necessary set the settle delay in Skanlt Software For more details refer to Settle delay on page 52 IMPORTANT Because of the relative nature of fluorescence it is recommended to use known samples or controls to verify instrument operation Fluorescence spectrum scanning The phases of the fluorescence spectrum scanning measurement are the same as for the fluorescence measurement but with a continuous range of wavelengths Refer to Fluorescence measurement on page 46 Absorbance measurement 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 Inthe absorbance 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 measur
51. ctuations 10 from nominal Installation category overvoltage category ll according to IEC 60664 1 see Note 1 Pollution degree 2 according to IEC 60664 1 see Note 2 Note 1 The installation category overvoltage category defines the level of transient overvoltage which the instrument is designed to withstand safely It depends on the nature of the electricity supply and its overvoltage protection means For example in CAT Il which is the category used for instruments in installations supplied from a supply comparable to public mains such as hospital and research laboratories and most industrial laboratories the expected transient overvoltage is 2500 V for a 230 V supply and 1500 V for a 120 V supply Note 2 The pollution degree describes the amount of conductive pollution present in the operating environment Pollution degree 2 assumes that normally only nonconductive pollution such as dust occurs with the exception of occasional conductivity caused by condensation Thermo Scientific Varioskan LUX 89 Table 7 16Error codes reported Thermo Scientific Troubleshooting IMPORTANT Do not use the instrument if it does not appear to function properly Error and warning codes When an error is detected the current operation is terminated After an error it is best to abort the current run and restart from the beginning after the problem is fixed The error Table 16 and warning codes lable 7 17 that may a
52. des all previous editions Remarks on screenshots and pictures The version number displayed in screenshots may not always be the one of the currently released version Screenshots and pictures are only replaced if the content related to the application has changed No liability for consequential damages Thermo Fisher Scientific shall not be liable for any indirect or consequential damages whatsoever arising out of the use or inability to use this product Power failure The system requires uninterrupted power supply in order to operate correctly Thermo Fisher Scientific has no responsibility whatsoever for system malfunctions arising from power failures Manufacturer Thermo Fisher Scientific Oy Ratastie 2 P O Box 100 Fl 01621 Vantaa Finland Thermo Scientific Thermo Scientific Preface About this guide Thermo Scientific Varioskan LUX is a modular multi technology microplate reader controlled by Thermo Scientific Skanlt Software for Microplate Readers This guide gives a thorough description of the instrument installation procedures and also recounts the main operations routine maintenance and troubleshooting of the instrument Related documentation In addition to this guide Thermo Fisher Scientific provides the following documents e Thermo Scientific Varioskan LUX User Manual Cat No N16044 e Thermo Scientific Skanit Software for Microplate Readers Technical Manual Cat No N16046
53. e painted surfaces to concentrated acids or alcohols for prolonged periods of time as damage may occur Plastic covers and surfaces can be cleaned with a mild laboratory detergent or ethanol If you believe that liquid has entered the Varioskan LUX first switch the instrument off Figure 2 2 and unplug the instrument Carry out corrective measures Refer to How to clean the measurement chamber on page 68 and Decontamination procedure on page 80 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 81 Although the Varioskan LUX 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 Varioskan LUX 67 Chapter5 Maintenance IMPORTANT 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 ElA tests resulting in inconsistent test results CAUTION If any surfaces have been contaminated with biohazardous material a mild sterilizing solution should be used IMPORTANT Do not autoclave any part of this instrument IMPORTANT Do not use alkaline or chlorite solutions for cleaning any parts of the measurement chamber Figure 3 21 w
54. e 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 A both inside and attached to the outside of the package in which you return your instrument or other items Varioskan LUX 81 Chapter5 Maintenance 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 85 for details on storage and transportation temperatures How to refit the transport locks When you relocate the instrument or ship it for service make sure you refit the transport lock of the tray holder Tray holder transport lock Note that the tray holder transport lock support piece is easily recognizable having a metallic color and a yellow label Figure 3 22 Refer to How to release the transport locks on page 31 work phases in reverse order to refitting Unfasten the attached tray from the screw fix of the tray holder Figure 3 25 Push the tray holder gently into the instrument by hand Lift up the dispenser sliding cover Figure 3 20 Be ON oL Remove the front cover by first pulling the two bolts one on each side at the bottom sideways then tu
55. ed 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 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 pack for service 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 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 15 and Figure 2 16 and the complete dispensing tube assemblies Figure 5 53 after decontamination e Install the transport lock of the tray holder Then replace the front cover by first inserting the two top door latches into place while at the same time turning the cover 30 and then snapping it shut Figure 5 46 e Pack the instrument according to the enclosed packing instructions e Use the original packaging to ensur
56. ement 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 Varioskan LUX 49 Chapter 4 Operations 50 Varioskan LUX 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 lf necessary set the settle delay in Skanlt Software For more details refer to Settle delay on page 52 Absorbance spectrum scanning The phases of the absorbance spectrum scanning measurement are the same as for the absorbance measurement but with a continuous range of wavelengths See Absorbance measurement on page 49 The air blank spectrum is also measured in absorbance spectrum scanning measurements Luminescence measurement In luminescence intensity measurements the following actions are carried out by the instrument 1 The tray is driven in 2 The LAT filter selector position is rotated to the blocked position
57. f a wavelength of light through a liquid log dimension 4 adapter The elevation device used to hold and raise the microplate in the tray AFP Alpha fetoprotein common label in FRET and TR FRET assays AlphaScreen AlphaScreen Amplified Luminescent Proximity Homogeneous Assay Screen is a versatile technology used to detect and quantify a broad range of analytes in complex biological processes AlphaLISA AlphaLISA extends AlphaScreen technology to allow for high throughput detection and quantification of markers in biomolecular reactions 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 B 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 C chemiluminescence Luminescence as a result of pure chemical reactions D 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 dif
58. fraction 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 disinfection The destruction of pathogenic bacteria usually with an antiseptic chemical or disinfectant Varioskan LUX 105 Glossary 106 Varioskan LUX double monochromator Two serially one after another connected 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 E 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 F 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
59. g measurement because this causes stray light to enter and aborts the measurement The dispenser sliding cover can however be left open Fluorescence measurement In fluorescence intensity Fl or time resolved fluorescence TRF measurements the following actions are carried out by the instrument 1 The tray is driven in 2 In fluorescence intensity measurements excitation and emission wavelengths are selected by rotating the excitation and emission monochromatic gratings On the other hand in time resolved fluorescence measurements the emission wavelengths are selected by rotating the filter wheel Thermo Scientific Figure 4 37 Dynamic range selection Thermo Scientific Operations Chapter 4 3 In the signal level calibration procedure the instrument reads the fluorescence from the reference chip 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 instrument uses the dynamic range setting the user has selected in the Skanlt Software measurement session Figure 4 37 AutoRange Manual range High range Medium high range Medium low range Low range The measured values are comparable regardless of the dynamic range selection AutoRange or any of the fixed manual ranges j High range Viedium h
60. gas concentration Varioskan LUX 65 Chapter 4 Operations CAUTION The module must only be used by trained and authorized personnel CAUTION Do not use cell or tissue cultures in the device that are not in accordance with the regulations of safety levels L1 L2 and L3 CAUTION Ensure that the working area is well ventilated In order to detect gas leakages CO and O sensors must be present on the wall of the room where the Varioskan LUX with a Integrated gas module is installed IMPORTANT The O sensor must be changed annually The disposable gas filters must also be changed during the same service or more frequently if necessary 66 Varioskan LUX Thermo Scientific Thermo Scientific Maintenance Contact local authorized technical service or your local Thermo Fisher Scientific representative for assistance if necessary IMPORTANT It is recommended to service the instrument at least yearly Preventive maintenance 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 IMPORTANT Painted surfaces can be cleaned with most laboratory detergents Dilute the cleaning agent as recommended by the manufacturer Do not expos
61. h the dispensing head located in the dispensing head holder and with a small vessel underneath 2 Either select the Wash function in Skanlt Software or use the Prime control button located on top of each dispenser Figure 2 5 and Figure 2 14 When you press the Prime control button continuously the instrument fills and then empties the syringe by 2096 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 3 Change the washing and rinsing solutions manually IMPORTANT Carefully insert the dispensing head straight into the dispensing head holder without damaging or contaminating the dispensing tip Thermo Scientific Varioskan LUX 61 Chapter 4 Operations Table 4 5 Compatibility chart of solvents suitable with the plastic materials used in the dispensers 62 Varioskan LUX Chemical resistance of the dispensers Table 4 5 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 IMPORTANT Failure to determine compatibility of chemicals used in individual applications with the dispensers may result in damage to the dispensers and o
62. he adapter however when you use microplates with lids To remove or replace the robotic tray adapter Figure 2 18 1 To remove the factory installed adapter unfasten the four adapter retaining screws fitted with washers Figure 2 18 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 18 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 A1 corner and fasten the A1 corner adapter retaining screw firmly Finally fasten the rest of the adapter retaining screws firmly to the tray Adapter for plate F PD ieee without lid 126 725 p AF Washer and adapter retaining screw Figure 2 18 Removing or replacing the adapter for plate w o lid 126 Refer to Table 2 2 and Chapter 8 Ordering Information Thermo Scientific Varioskan LUX 27 Thermo Scientific Installation CAUTION The Varioskan LUX weighs 53 to 60 kg 117 131 Ibs depending on the configuration and care must be taken when lifting it Two persons must lift the instrument one on each side by hooking their fingers under the sides What to do upon delivery This section covers the relevant procedures to be carried out upon arrival of the instrument How to unpack Move the packed instrument to its site of o
63. he emission filters are missing or they are misplaced in the module in relation to the software settings Check that the filters are present in the correct filter slots CO concentration below target The CO gas cylinder closed Open the CO gas cylinder Check that all the gas lines are properly connected Thermo Scientific Table 8 19 Instrument catalog numbers Thermo Scientific Ordering Information Contact your local Thermo Fisher Scientific representative for ordering and service information Table 8 19 through Table 8 25 Varioskan LUX VLOOOODO Varioskan LUX with fluorescence top and absorbance VLOLOODO Varioskan LUX with fluorescence top absorbance and luminescence VLOLOTDO Varioskan LUX with fluorescence top absorbance luminescence and TRF VLOLAODO Varioskan LUX with fluorescence top absorbance luminescence and AlphaScreen VLOLATDO Varioskan LUX with fluorescence top absorbance luminescence AlphaScreen and TRF VLBOOODO Varioskan LUX with fluorescence top amp bottom and absorbance VLBLOODO Varioskan LUX with fluorescence top amp bottom absorbance and luminescence VLBLOTDO Varioskan LUX with fluorescence top amp bottom absorbance luminescence and TRF VLBLAODO Varioskan LUX with fluorescence top amp bottom absorbance luminescence and AlphaScreen VLBLATDO Var
64. he transportlocKs i e I 82 Tray holder transport lock 6 IR 82 Chapter 6 Technical Specifications 0 ccc cece e eee eee eee nmn 85 General Specifications s p ke bie wk aa dee dewe aaron STA 85 Performance specifications in room temperature n naana 86 Safety SPSCMCATONG a author ae A Tace ae ON a a URS ee YS 89 In conformity with the requirements 0 00 ccc cece eect teens 89 Chapter 7 Troubleshooting ccce cece cece eee eee eee n n m Hn mH mm 91 Error and Warming codes HH 91 Troubleshooting gE eseni doute Re ago acie e eo rie tee ee tetas emet denda 96 Chapter8 Ordering Information llilellseeeseenn nnn KR 97 PEINT 97 Gas module for Varioskarn LUX sista re icc ca cicc oet a ACER ERROR e o ee 98 Upgrade kits for Varioskan LUX nesai iiis 98 Accessories for Varioskan LUX isses teen eee 98 Verification tools for Varioskan LUX m teens 99 Skant SOWA E sc aas ree OP ELO tq RTL dee d c RERO D RC ep et A 99 Filters for Varioskan LUX iiis RR HH 99 Appendix A Certificate of Decontamination 000c cece cece eee eee eens 101 Appendix B Preparation of Luminescence Alignment Plate luueeeesee 103 Required reagents materials and equipment 00 ee ete 103 POGUELCOMPONEMIS 53 4 230s bh teet ey eee e dere E ken Or ee 103 Reconstitution of assay components 6 ee teen n 104 PIDETCIMOPIMSEMUCTIONS aa eo ibas 104 3l08Sdl
65. hich may result in immediate damage to the instrument How to clean the measurement chamber 1 Switch the Varioskan LUX off by turning 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 20 3 f you suspect that liquids have entered the measurement chamber Figure 5 47 remove the front cover by first pulling the two bolts 1 one on each side at the bottom sideways then turning the bottom of the cover 2 30 and finally by lifting it off according to Figure 5 46 Figure 5 46 Front cover removed 68 Varioskan LUX Thermo Scientific Figure 5 47 Internal view of the measurement chamber Thermo Scientific Maintenance Chapter 5 If you have spilt infectious agents into the measurement chamber Figure 5 47 decontaminate according to Decontamination procedure on page 80 Otherwise clean the measurement chamber surface using a soft cloth or tissue paper soaked in a mild detergent solution soap solution or 70 ethanol IMPORTANT Do not leave or store corrosive materials inside the measurement chamber AL IMPORTANT Do not touch the photometric or the bottom reading optics lens with bare hands on the top 5 Replace the front cover by first inserting the two top door latches into
66. igh range Viedium low range ow range T U Concentration decades 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 Tip Do not adjust the dynamic range selection if you do not know which dynamic range to use AutoRange is almost always the optimal reading range 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 if the settle delay is off Sometimes it is necessary to set the settle delay on in Skanlt Software For more details refer to Settle delay on page 52 Varioskan LUX 47 Chapter 4 Operations 48 Varioskan LUX 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 on the other hand produces 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 measureme
67. ilure is repeated contact authorized technical service All error messages are stored in the internal memory log file of Varioskan LUX The error log file can be accessed with Skanlt Software by selecting Settings gt Instrument gt Edit instrument parameters gt Reports gt Instrument Error Log gt Run Report Refer to Error and warning codes on page 91 Loading the microplate IMPORTANT Ensure that you select a correct plate type Too high a plate may become jammed and with too low a plate the dispensing might fail and skip over Also with too low a plate the plate may not be at the optimal height for measurement Note When placing a microplate onto the tray always make sure the correct plate type has been selected in Skanlt Software Plate template before you do anything else To load the microplate 1 Ensure that the plate type tray adapter and the Skanlt Software plate template match Refer to Plate trays and adapters on page 22 and How to install the plate tray and adapter on page 33 44 Varioskan LUX Thermo Scientific Figure 4 36 Microplate loaded Thermo Scientific Operations Chapter 4 Microplate Tray Adapter Plate template see Plate trays and adapters see the Skanlt Software on page 19 Technical Manual 2 The tray is driven out after which the microplate can be loaded If the tray is in first drive the tray out by pressing the Plate Out button or by selecti
68. inge 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 2 5 000 yl with 1 ul increments 5 ml syringe 5 25 000 pl with 5 ul increments Automatic safety control based on maximum well volume 1 ml syringe 0 40 mm tip size default Accuracy lt 1 ul with 50 ul Precision lt 1 pl with 50 ul 1 ml syringe Accuracy lt 0 2 ul with 5 ul 0 25 mm tip size Precision lt 0 25 ul with 5 ul Dispensing speed 96 well plate 30 s 384 well plate 80 s 5 ul well 1 ml syringe 0 40 mm tip Dead volume 100 ul total tubing volume 800 pl Tip priming volume 2 10 ul Plate sensing Photometric Plate types 6 384 well plates Thermo Scientific Technical Specifications Chapter 6 Safety specifications This section describes the safety specifications for the Varioskan LUX instrument In conformity with the requirements Varioskan LUX bears the following markings Type 3020 100 240 Vac 50 60 Hz 200 VA CE marking cTUVus 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 2000 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 flu
69. ioskan LUX with fluorescence top amp bottom absorbance luminescence AlphaScreen and TRF VLOOOOD1 Varioskan LUX with fluorescence top and absorbance 1 dispenser VLOLOOD1 Varioskan LUX with fluorescence top absorbance and luminescence 1 dispenser VLOLOTD1 Varioskan LUX with fluorescence top absorbance luminescence and TRE 1 dispenser VLOLAOD1 Varioskan LUX with fluorescence top absorbance luminescence and AlphaScreen 1 dispenser VLOLATD1 Varioskan LUX with fluorescence top absorbance luminescence AlphaScreen and TRF 1 dispenser VLB000D1 Varioskan LUX with fluorescence top amp bottom and absorbance 1 dispenser VLBLOOD1 Varioskan LUX with fluorescence top amp bottom absorbance and luminescence 1 dispenser VLBLOTD1 Varioskan LUX with fluorescence top amp bottom absorbance luminescence and TRF 1 dispenser VLBLAOD1 Varioskan LUX with fluorescence top amp bottom absorbance luminescence and AlphaScreen 1 dispenser VLBLATD1 Varioskan LUX with fluorescence top amp bottom absorbance luminescence AlphaScreen and TRF 1 dispenser Continued Varioskan LUX 97 Chapter 8 Ordering Information Table 8 20 Catalog number for Integrated gas module Table 8 21 Catalog numbers for upgrade kits Table 8 22 Catalog numbers for accessories 98 Varioskan LUX Cont VLO000D2 Varioskan LUX with fluorescence top and absorbance 2 dispensers LOLOOD2 Varioskan LUX with fluorescence top
70. istance between measurement points is too short Use normal measurement instead of scan Do for scan measurement The plate cannot be moved as not try to measure all the points with a single slowly as the distance requires scan but use interleaving scans Continued Varioskan LUX 91 Chapter 7 Troubleshooting 92 Varioskan LUX Cont Code Explanation Suggested action 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 a shorter sampling time the sampling time requires 19 The requested plate position is outside the mechanical Check the plate definition PLA command limits of the XY table Remember that the actual plate position depends on the measurement method and dispensers used Note that plate scanning SCA command requires some extra space in the scanning direction for start stop ramps 20 The offset voltage of the temperature measurement Contact service electronics is too high 21 An offset voltage on the QUAD analog board is too high Contact service 22 A background noise on the QUAD analog board is too Contact service Commands REP 21 and REP high 38 may be used to diagnose the source of this error 23 Error when checking the QUAD analog board reference Contact service voltage 24 Calibration of the Q
71. ize 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 2 Use detergent in the well if possible Set on the settle delay in Skanlt Software The used settle delay time is automatically selected according to the plate format o Settle delay is the time for waiting for the liquid surface to settle before the reading is carried out e Settle delay times are dependent on the plate format Table 4 3 Table 4 35ettle delay times vs plate Plate format Settle delay time ms formats 6 well plates ii 12 well plates 1300 24 well plates 200 48 well plates 96 well plates 108 384 well plates 5n 1536 well plates 2 For more information on the settle delay refer to the Skanlt Software Technical Manual 52 Varioskan LUX Thermo Scientific Thermo Scientific Operations Chapter 4 Measurements under controlled atmosphere In measurements under controlled atmosphere where the user wants to follow a long term process with cultivated cell cultures the following actions are carried out 1 The user sets the desired gas concentration using Skanlt Software Wait till the concentration has been reached Adjust either the CO or the O concentration or both to the desired level 2 The user sets the desired temperature using Skanlt Software Wait till the temperature has been reached 3
72. jo o Oo pjojo o Gas control The instrument can be equipped with an optional Integrated gas module to achieve a controlled atmosphere inside the instrument The module consists of CO and O sensors valves and the gas supply system The sensors are placed in a measurement chamber The CO and O concentrations are controlled independently by tuning the valves according to the readings of the Sensors Atmosphere In the space of the instrument the particular physiological ambient conditions for the preparation and cultivation of cell and tissue cultures are simulated The atmosphere is determined by the following factors CO concentration and 0 concentration CO supply To ensure the growth conditions for the cell and tissue cultures the instrument should be supplied with CO The pH of bicarbonate buffered culture media largely depends on the CO content of the atmosphere The CO content of the atmosphere can be controlled within a range of 0 1 to 15 The CO supply must have either of the following quality characteristics Purity 99 5 min e Medical gas quality Thermo Scientific Operations Chapter 4 N supply If the oxygen content during operation is to be lowered to less than 21 air oxygen content the instrument is supplied with nitrogen The O content of the atmosphere can be controlled within a range of 1 to 21 Use The Integrated gas module is a laboratory device for prepa
73. kan LUX Thermo Scientific Figure 2 5 Varioskan LUX internal views Thermo Scientific Internal view Functional Description Chapter 2 The internal view of the Varioskan LUX instrument is shown in Figure 2 5 The blue color alerts the user to find units parts that may be handled Plate In Out button LAT module Prime and Empty buttons Dispensers 1 and 2 Reagent bottle holder SONS Male connector of the dispensing tube assembly 3 Dispenser Reagent bottle holder Aspirate tube Varioskan LUX 15 Chapter2 Functional Description Figure 2 6 Varioskan LUX optics and measurement positions F and L 16 Varioskan LUX Optical system The Varioskan LUX uses Fluorescence intensity Time resolved fluorescence Luminescence Absorbance and AlphaScreen measurement techniques Fluorescence intensity measurements are conducted from the top or bottom of the well luminescence AlphaScreen and time resolved fluorescence measurements from the top of the well and absorbance measurements through the well The principle of the Varioskan LUX 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
74. lace a dispenser syringe of 1 ml Figure 5 54 or 5 ml 1 Remove the liquid from the dispenser syringe Figure 5 53 and from the tubing 2 Switch off the power from the instrument by turning the mains switch into the off position Figure 2 2 3 Loosen the plunger lock screw Figure 5 53 approximately three full turns clockwise Figure 5 54 item c 4 Pull the plunger holder arm Figure 5 53 firmly down Figure 5 54 item b 5 Unscrew the syringe from the valve Figure 5 54 item a 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 54 Thermo Scientific Varioskan LUX 77 Chapter5 Maintenance Syringe Plunger Holder Figure 5 54 Replacing the dispenser syringe f Plunger Lock Screw Valve Syringe Define the syringe size in Skanlt Software if you have changed the size Replacing the 3 port valve To replace the 3 port valve Figure 5 53 and Figure 5 55 it is recommended that you contact authorized technical service 1 2 3 78 Varioskan LUX Remove the fluid from the dispenser s Remove the syringe and tubing Remove the two Phillips head screws on the front of the valve and then remove the valve from the dispenser s Install the new valve by placing it on the front panel so the screw holes
75. le Refer to Emission reading module on page 19 The absorbance measurement module measures light beam intensity passing through the well Absorbance measurement is carried out by using the excitation optics module as the absorbance measurement light source Refer to Absorbance measurement module on page 20 Fluorescence 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 Fluorescence bottom reading on page 20 The LAT module consists of Luminescence measurement capability Optionally it may also contain AlphaScreen and or Time resolved fluorescence measurement capabilities The LAT module is an option that is mounted in the front of the unit with the dispensers to the right The LAT measurement optics module collects with a wide angle emitted light that is produced in the microplate well and measured by a photomultiplier tube The wavelengths are differentiated by filters not monochromators The shutter and filter selector incorporates positions for open blocked and eight configurable filters Refer to LAT measurement module on page 21 The LAT module has a capability to measure spectral Luminescence and Time resolved fluorescence For these measurement modalities the Spectral scanning module is used instead of LAT module photo multiplier tube Excitation optics Excitation optics Figure 2 7 consists of
76. luorescein well black 384 well plate 6 decades Bottom reading 4 fmol fluorescein well clear bottom black 384 square well plate gt 5 5 decades Measurement time Fluorescence intensity 10 1000 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 2 2 s well 400 500 nm 1 flash 2 nm steps Plate types 6 1536 well plates top reading 6 384 well plates bottom reading 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 2nm Linear measurement range 0 4 Abs 96 well plate at 450 nm 2 0 3 Abs 384 well plate at 450 nm 2 Absorbance resolution 0 001 Abs Accuracy 0 003 Abs or 2 at 200 399 nm 0 2 Abs 0 003 Abs or 1 at 400 1000 nm 0 3 Abs Precision SD 0 001 Abs or CV 0 596 at 450 nm 0 3 Abs Stray light 0 00596 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 2 2 s well 400 500 nm 1 flash 2 nm steps
77. ng Run plate out in Skanlt Software 1 Microplate 2 Plate tray Load the microplate onto the plate tray for measurement Figure 4 36 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 15 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 A1 corner is positioned in the top left corner of the tray Figure 4 36 The tray frame is marked with A1 to facilitate correct insertion of the microplate Select the measurement parameters to define the measurement After this the measurement is executed with Skanlt Software Refer to the Skanit Software Technical Manual Varioskan LUX 45 Chapter 4 Operations 46 Varioskan LUX Automatic runtime calibration 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 Note If any measurement is performed after a specific time 10 minutes or an hour
78. ng range 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 principle for achieving best sensitivity is to select the lowest possible range to prevent 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 6 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 7 f necessary set the settle delay in Skanlt Software For more details refer to Settle delay on page 52 Luminescence spectrum scanning The phases of the luminescence spectrum scanning measuremen
79. ng universal tray The microplate Figure 4 41 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 4 41 The tray and the upper heater together form the isothermal chamber The upper element is slightly warmer than the lower element to avoid condensation on the plate lid Microplate 96 well adapter for plate without lid 2 Bottom reading hole Universal tray frame Note The samples in the microplate reach the target temperature usually much later than the instrument Varioskan LUX 55 Chapter 4 Operations Figure 4 42 Varioskan LUX dispensing system 56 Varioskan LUX Dispensers The instrument is equipped with up to two optional dispensers Figure 2 5 for automatic and accurate reagent addition The dispensers are located in the instrument housing Figure 2 2 under the dispenser sliding cover and consist of a pump with a valve a syringe 1 ml tubing and a dispensing head Figure 4 42 The instrument supports simultaneous dispensing and reading enabling fast signal monitoring from the very start of the reaction Syringe The 1 ml syringe is the default syringe A 5 ml syringe is available on request The volume range and increment depend on the syringe size used The volume range is from 2 ul to 5000 ul in 1 ul increments with the 1 ml syringe and from 5 ul to 25 000 ul in 5 ul increme
80. nits Optimizing the measurement position for each unit will ensure the specified crosstalk performance of the instrument The LAT module is fully functional and can also be used without this luminescence 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 adjacent wells in luminescence To execute the alignment sequence a white 384 well plate with 30 ul of luminous solution in well H12 is needed The luminescence intensity of the solution should be in the range of 10 to 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 Thermo Scientific Installation Chapter 3 Detailed instructions on how the alignment plate can be prepared using Promega s ENLITEN ATP kit are given in Appendix B Preparation
81. ns 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 The wavelength selection for Luminescence AlphaScreen and Time resolved fluorescence measurements are made with a set of filters LAT excitation module produces excitation light for AlphaScreen and Time resolved Fluorescence measurements by a light emitting diode and a xenon flash lamp respectively Measurement of excitation light in spectral scanning Luminescence and Time resolved fluorescence is performed by Spectral scanning module Varioskan LUX 21 Chapter2 Functional Description Figure 2 14 Control buttons 22 Varioskan LUX Control buttons There are five control buttons e One blue Plate In Out button for driving the plate carrier in or out Plate In Out function Figure 2 14 and e Two blue Prime and Empty buttons for priming and emptying the dispenser tubing Prime Empty function of each dispenser Figure 2 14 Note that the Prime and Empty buttons are only present if there is a dispenser fitted 1 Plate In Out control button 2 Prime control buttons 3 Empty control buttons Plate trays and adapters There are two types of plate trays universal and robotic
82. nt 5 The wells are measured with a selected measurement time that can vary from 10 to 1000 ms in fluorescence intensity measurements and from 10 to 10 000 ms in time resolved fluorescence TRF 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 it is necessary 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 measurement parameters TRF delay time and TRF integration time The TRF delay time defines the time difference between the excitation flash and the start of emission signal collection while the TRF integration time defines the time used for emission signal collection When the Varioskan LUX performs a TRF measurement it excites the sample with a very short light pulse waits for the defined TRF delay time and then colle
83. ntific www thermoscientific com 2015 Thermo Fisher Scientific Inc All rights reserved Decon 90 is a trademark of Cole Palmer Microcide SQ is a trademark of Global Biotechnologies Inc Microsoft and Windows are trademarks of Microsoft Corporation Virkon is a trademark of E I du Pont de Nemours and Company or its affiliates All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Africa Other 27 11 570 1840 Finland Norway Sweden Middle East 43 1 333 50 34 0 Australia 61 2 8844 9500 46 8 556 468 00 Netherlands 31 76 579 55 55 Austria 43 1 333 50 34 0 France 33 1 60 92 48 00 South Africa 27 11 570 1840 Belgium 32 53 73 42 41 Germany 49 6103 408 1014 Spain 34 914 845 965 Canada 1 800 530 8447 India 91 22 6742 9434 Switzerland 41 61 716 77 00 China 86 10 8419 3588 Italy 39 02 950 591 UK 44 1442 233555 Denmark 45 70 23 62 60 Japan 81 45 453 9100 USA 1 800 532 4752 Europe Other 43 1 333 50 340 Latin America 1 608 276 5659 N16045 Varioskan LUX Technical Manual Thermo SCIENTIFIC A Thermo Fisher Scientific Brand
84. nts with the 5 ml syringe The upper limit of the range is 80 of the maximum well volume which is supported by Skanlt Software when using multiple dispensings into the same well Reagent consumption 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 4 42 and the tip priming feature ensures that even very small volumes can be dispensed accurately Male connector of the dispensing head position sensor Dispensing tube assembly Dispenser syringe and plunger Plunger lock screw Valve Aspirate tube assembly Dispensing head Dispensing tip WwW NY CON O 5 Thermo Scientific Operations Chapter 4 Dispensing tip There are two different dispensing tip options Figure 4 43 e The black dispensing tip 0 40 0 40 mm and e The transparent dispensing tip 0 25 0 25 mm Dispensing tip 0 40 Cat No NO3080 is the default tip and is recommended for volumes gt 5 ul since dispensing is then more rapid Dispensing tip 0 25 Cat No NO3081 has to be ordered separately and is intended for small volumes in the range of 2 to 20 ul ow oze Figure 4 43 Dispensing tip options 0 40 mm and 0 25 mm
85. ode luminescence 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 Thermo Scientific Operations Chapter 4 IMPORTANT Do not use any liquids with automatic dispensers that can cause precipitation or coagulation or that contain any mechanical particles You may need to adjust the dispensing speed The default setting is for water You can find the adjustments and selections in Skanlt Software Emptying The dispensers can be emptied either manually by using the control buttons for emptying the dispensers or by clicking the Empty icon in Skanlt Software To avoid wasting 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 The total dispenser tubing volume is 800 ul The dead volume reagent loss is 100 yl To empty the dispenser 1 Keep the aspirate tube in the reagent bottle 2 Press the Empty button until you have emptied all the liquid into the reagent bottle 3 Remove the aspirate tube carefully from the reagent bottle Dispenser washing The dispensers can be washed using Skanlt Software or the control buttons for priming the dispensers Wash the dispenser tubing if necessary Refer to the washing instructions in the Skanlt Software Technical Manual To wash the dispenser 1 Carry out dispenser washing wit
86. peration To prevent condensation the instrument should be left in its protective plastic wrapping until the ambient temperature has been reached Unpack the Varioskan LUX 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 for Varioskan LUX reader e Packing list for Varioskan LUX reader e Warranty Certificate card e Varioskan LUX reader performance measurements IMPORTANT Do not touch or loosen any screws or parts other than those specifically allowed in the instructions Doing so might cause misalignment and will void the instrument warranty 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 81 Varioskan LUX 29 Chapter 3 Installation 30 Varioskan LUX Checking delivery for completeness Check the enclosed packing list against order If any parts are missing contact your local Thermo Fisher Scientific representative or Thermo Fisher Scientific Oy
87. peration commands for some measurement methods 109 The interval time defined with a SYN command for the next two measurements was too short The second measurement started later than requested 2 110 114 Dark level interpolation for luminescence results was requested but the results buffer became full before it could be applied Set gas concentration not reached Do not use dark level interpolation or use it between each well Alternately try reducing the well group size The instrument cannot apply dark level interpolation if more than 1 536 results are measured between the dark level measurement points See command SET 11 112 Gas concentration out of the set value The following commands will not execute if there is such a start up error that there is a possibility for an incorrect measurement result CAL DIS DME MEA PRI PTI and SCA Also the command FCA executes but will fail at the end with error 81 unless it fails earlier to some other error Error 81 is persistent it can be cleared to execute other commands than listed above but the listed commands return the same error again A successful start up sequence is required to clear the error permanently 2 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 Thermo Scientific Varioskan LUX 9
88. place while at the same time turning the cover 30 and then snapping it shut Figure 2 2 and Figure 5 46 Also close the dispensing sliding cover arrows in the opposite direction than in Figure 3 21 How to clean the optics in the measurement chamber To clean the optics that is the absorbance lens and the fluorescence bottom reading optics 1 es Soak a lint free tissue in 70 ethanol Wipe over the lens in a straight movement from inside to out Do not use any circular movements when cleaning the optics Repeat the procedure if the lens is not clean IMPORTANT Do not touch the optical lenses with bare hands Avoid touching the lens of the absorbance window Figure 5 47 and the white reference chip with bare hands Varioskan LUX 69 Chapter5 Maintenance 70 Varioskan LUX How to clean the tray To clean the tray follow the instructions below Keep the instrument tray s Figure 2 15 and Figure 2 16 clean to avoid dust and dirt from entering the measurement chamber Figure 5 47 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 If you have spilt infectious agents on the tray decontaminate according to Decontamination procedure on page 80 CAUTION Ensure that the bottom of each mic
89. ppear in Skanlt Software are presented below Code EME LOU Suggested action 0 The command was executed successfully i Internal firmware error When this error occurs Contact service the internal software halts This error can only be seen in the error log returned for the LOG command 2 The instrument did not recognize the command it Contact service received 3 The arguments of the received command are not valid Contact service 4 The XY table X position is incorrect Contact service 5 The XY table Y position is incorrect Contact service 6 The 1 excitation grating position is incorrect Contact service 1 The 2 excitation grating position is incorrect Contact service 8 The 1s 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 polarization filter above below selector Contact service position is incorrect 13 Emission polarization filter above below selector Contact service position is incorrect 14 Luminescence filter position is incorrect Contact service 15 Excitation bandwidth selector position is incorrect Contact service 16 Beam diameter selector position is incorrect Contact service 17 The d
90. r Varioskan LUX 75 Chapter5 Maintenance 2 Replace the dispensing tip connected with a small piece of silicone tube in the dispensing tube Figure 5 52 Use the dispensing 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 Figure 5 52 Replacing the dispensing ip B tip B 1 Dispensing head tube 2 Dispensing tip to be discarded 3 Replace the dispensing head tube Figure 5 52 Fasten the parts by turning in the opposite directions than shown in Figure 5 51 4 Change the tip size setting in Skanlt Software if you have changed the tip size Replacing a dispenser syringe The dispenser syringe replacement procedure is carried out in two stages Manual replacement of the dispenser syringe Defining the dispenser syringe data using Skanlt Software only if you have changed the syringe size If the instrument has the wrong information about the syringe used it will dispense incorrect volumes Refer to the Skanlt Software Technical Manual 76 Varioskan LUX Thermo Scientific Maintenance Chapter 5 Figure 5 53 Dispenser assembly Male connector of the dispensing head position sensor Dispensing tube assembly Dispenser syringe and plunger wh 4 Plunger lock screw 5 Valve 6 Aspirate tube assembly 7 Dispensing head 8 Dispensing tip To rep
91. r parameters to make room for new command parameters 72 The drop shield position is incorrect Contact service 73 Emission first bandwidth selector position is incorrect Contact service 74 Emission second bandwidth selector position is incorrect Contact service 75 Default PMT voltages calibration failed Contact service 76 Expanded dynamic range PMT voltages calibration failed 17 The dark level signal on some of the QUAD board input channels is too high Contact service Contact service 78 PMT linearity calibration failed 79 Bandwidth factors calibration failed Contact service Contact service Continued Varioskan LUX 93 Chapter 7 Troubleshooting 94 Varioskan LUX Cont Code Explanation Suggested action 80 Measurement chamber door is open See that nothing is obstructing the measurement chamber door from closing when the plate is driven in Do not open the door while the instrument is measuring 81 Any error during start up preventing the execution of The measurement chamber door must be some commands closed during start up If it is not contact service 82 The number of filter pairs does not match the number of Make sure that the number of filter pairs in a delay integration time pairs This error may be reported FIL command and the number of delay in response to a measurement command when a TRF in
92. r test results 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 Daikin Note Check 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 Classification in the table No data available No effect excellent Minor effect good Moderate effect fair Severe effect not recommended Polypropylene satisfactory to 22 C 72 F Polypropylene satisfactory to 49 C 120 F 0N Teflon Polypropylene Acetaldehyde 0 0 0 Acetates 0 0 Acetic acid 0 0 0 ic anhydride 0 Acetone 0 0 0 Acetyl bromide 0 Ammonia 0 0 Ammonium acetate 0 Ammonium hydroxide 0 0 0 Ammonium phosphate 0 0 Continued Thermo Scientific Thermo Scientific Cont Solvent Ammonium sulfate Teflon Operations Chapter 4 Polypropylene 0 Amyl acetate Aniline Benzene 1O
93. required filters can be used The LAT module has built in excitation filters for TRF and AlphaScreen The TRF and AlphaScreen and luminescence emission filters you need to install yourself Note All measurement technologies except AlphaScreen allow spectral scanning measurements with monochromators The instrument has an incubator for temperature control up to 45 C and a plate shaking capability with orbital shaking mode The instrument can also be equipped with Dispensers up to two for automatic reagent addition e Integrated gas module for controlling the gas atmosphere CO amp 0 inside the instrument The optical system of the instrument allows you to perform Fluorescence intensity measurements from the top or bottom of the well Luminescence TRF and AlphaScreen measurements from the top of the well Absorbance measurements through the well Note Fluorescence measurements from the bottom of the well require an instrument model supporting bottom reading Note Your instrument may not have all of the features presented in this guide As the instrument is modular you can upgrade it with missing features later 10 Varioskan LUX Thermo Scientific Thermo Scientific Introduction to Varioskan LUX Chapter 1 Intended use The Varioskan LUX modular multi technology reader Figure 1 1 is used to measure fluorescence intensity absorbance luminescence time resolved fluorescence and AlphaScreen from samples
94. ring and cultivating cell and tissue cultures The device allows the simulation of the special physiological ambient conditions for these cultures due to the exact control of CO content and e OJN content The Integrated gas module e Offers a comprehensive solution for a variety of cell based applications such as cell toxicity cell proliferation and stem cell cultivation e Can be used in conjunction with all measurement technologies e Enables simultaneous independent control of CO and O concentrations e CO concentration is generally adjusted to ca 5 6 e 0 concentration is adjusted with nitrogen N To use the Integrated gas module 1 Turn on the instrument and open Skanlt Software 2 Open the CO or N gas cylinder and ensure that the operating pressure is correct 3 Set the desired gas concentration using Skanlt Software a Click the gas atmosphere icon above the Start button to open the pop up window Figure 5 45 b Set the O and CO levels as needed and click OK You can now see the current and target gas concentrations above the Start button Incubator Off 20 0 C O 2 CO 2 5 0 m Regulate oxygen Oz level 25 21 0 v Vv Regulate carbon dioxide CO level 75 5 0 uy Figure 5 45 The gas atmosphere pop up o concer window Thermo Scientific 4 Wait till the concentration has been reached The time required for this depends on the used gas type and the
95. rning the bottom of the cover 30 and finally by lifting it off according to Figure 5 56 Al MY 82 Varioskan LUX Thermo Scientific Figure 5 56 Transport lock released A and fastened B screws 1 4 shown Thermo Scientific Maintenance Chapter 5 Ii 10 Remove the two screws marked 1 and 2 of the transport lock from their storage site on the track mechanism bar Figure 5 56 A Remove the screw marked number 3 Figure 5 56 A Loosen the screw marked number 4 of the transport lock slightly by using the hexagonal screwdriver supplied Figure 5 56 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 56 B Pull the tray holder gently from inside the instrument so that you can fit the transport lock bar into its groove in the tray holder Refit the two screws marked 1 and 2 into the holes designated for them using the hexagonal screwdriver supplied Figure 5 56 B At the same time replace
96. roplate is dry Fluid on the bottom of a microplate may constitute a contamination hazard Use proper laboratory practices when handling any hazardous materials Keep all the holes in the adapter clean both the identification holes Figure 2 15 and the well holes Table 2 1 and Table 2 2 IMPORTANT Do not autoclave the tray Figure 2 15 and Figure 2 16 How to clean the reagent basin and dispensing area To clean the reagent basin and dispensing area follow the instructions below CAUTION If any surfaces have been contaminated with biohazardous material a mild sterilizing solution should be used Keep the reagent basin Figure 2 5 and dispensing area Figure 3 29 clean Clean the surface daily using a soft cloth or tissue paper soaked in a mild detergent solution soap solution or 7096 ethanol Wipe up spills immediately Do not use formaldehyde How to clean the LAT module 1 Switch off the Varioskan LUX by turning 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 LAT module according to Figure 5 48 Thermo Scientific Maintenance Chapter 5 Figure 5 48 Removing the LAT module Lift open the cover 2 Detach the two connectors Unfasten the two bottom screws Liftthe module off upwards
97. rument before disposal Refer to Decontamination procedure on page 80 about decontamination Note Observe all federal state and local environmental regulations Follow laboratory and country specific procedures for biohazardous or radioactive waste disposal All device components can be disposed of after they have been decontaminated properly 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 Thermo Scientific Varioskan LUX 79 Chapter5 Maintenance 80 Varioskan LUX Pollution degree 2 see Safety specifications on page 89 Method of disposal Electronic waste Contaminated waste Infectious waste Do not treat electrical and electronic equipment as unsorted waste Collect waste from electrical and electronic equipment separately Regarding the original packaging and packing materials use the recycling operators known to you Note Thermo Fisher Scientific offers a recycling service for discarded components at the Owner s expense For more information contact your local Thermo Fisher Scientific representative Decontamination procedure If you have spilled infectious agents carry out the decontamination procedure CAUTION The decontamination procedure should be performed by authorized trained personnel in a well ventilated room wearing disposable gloves prote
98. s Alpha measurement emission wavelength range 400 660 nm Alpha measurement limit of detection lt 100 amol phosphotyrosine well white 384 well plate Performance specifications TRF Light source Xenon flash lamp TRF excitation wavelength 334 nm Emission wavelength selection Filter wheel with up to 8 optional filter positions TRF emission wavelength range 400 670 nm Eu sensitivity for 384 well plate 1 amol well white low volume 384 well plate dynamic range gt 6 decades Performance specifications Measurement chamber O concentration accuracy Integrated Gas Module x 1 096 37 C 1 0 Measurement chamber CO concentration accuracy 1 0 37 C 5 CO Measurement chamber O 1 21 concentration Measurement chamber CO 0 1 15 concentration Gas concentration recovery time 10 minutes Varioskan LUX 87 Chapter 6 Technical Specifications Table 6 13 Incubator Table 6 14 Shaker Table 6 15 Dispensers 88 Varioskan LUX Performance specifications Incubator Temperature range From ambient 4 C to 45 C Setting range From 10 C to 45 C in 0 1 C increments Liquid warm up time 1 h from 25 C to 37 C covered 96 well plate 200 ul water well Shaking method Orbital shaking Shaking speed 60 1200 rpm Maximum centrifugal force 1G Performance specifications Dispensers Syr
99. s Figure 3 28 it is important that the recommended cleaning maintenance instructions are followed Refer also to Dispenser washing on page 61 For use of the Wash functionality refer to the Skanlt Software Technical Manual The Varioskan LUX 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 Thermo Scientific Thermo Scientific Maintenance Chapter 5 Daily maintenance The basic maintenance procedure should be performed regularly and on a daily basis to ensure proper dispenser operation 1 Flush the dispenser tubings Figure 3 30 out thoroughly with distilled deionized 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 IMPORTANT Do not allow the dispensers to run dry for more than a few cycles Weekly maintenance 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 Remove the dispensing heads Figure 2 5 from the dispensing head positioners Figure 2 5
100. start the measurement simultaneously to dispensing without any delay time between them place the dispensing head into the dispensing position which points at the correct measurement technology For example flash luminescence measurements require simultaneous measurement and dispensing However both dispensers and dispensing positions can be used to dispense into any well using any measurement mode s If you are using a dispensing position which does not point at the measurement position of the correct measurement technology an extra plate movement may be carried out before the measurement step This may cause minor time delays Measurement position F Measurement position L absorbance luminescence fluorescence top of the well fluorescence bottom of the well spectral TRF TRF spectral luminescence AlphaScreen Thermo Scientific Operations Chapter 4 The dispensers are each equipped with a dispensing head position sensor Figure 4 44 and Figure 3 30 The instrument recognizes a dispensing head placed into the dispensing position Dispensing will not take place 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 set up the dispensers on
101. substance which emits light at room temperature A group of atoms that can make a compound luminescent M 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 0 optical density absorbance log 1 transmittance log dimension 0 D 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 P 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 Glossary 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 Q quadruple monochromator Dual serially connected double monochromators quantum yield Q The ratio of the number of emitted photons to the number of excited molecules
102. t are the same as for the luminescence measurement but it always uses the spectral scanning module monochromators for controlling the wavelengths Refer to Luminescence measurement on page 50 Thermo Scientific Varioskan LUX 51 Chapter 4 Operations AlphaScreen measurement In AlphaScreen measurements the following actions are carried out by the instrument ll If not already installed install the AlphaScreen AlphaLISA filter before starting the session 2 The tray is driven in 3 The filter selector position is rotated to the selected AlphaScreen AlphaLISA position 4 In Steps 4 7 the instrument uses the settings the user has selected in the Skanlt Software session The excitation time is selected 5 The delay time is selected 6 The integration time is selected 7 The measurement time ms equals the sum of the excitation delay and integration times It is shown on the user interface but is not user selectable The time can vary from 20 to 1000 ms Settle delay 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 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 optim
103. t start up for example nstrument configuration Tray positioning Measurement selectors positioning Excitation and emission double monochromators Excitation and emission diffraction order filters Non volatile memory Varioskan LUX 43 Chapter 4 Operations e Temperature measurement electronics e Measurement electronics e Excitation bandwidth selector e Light sources e Reference detectors e LAT filter positioning e LAT crosstalk shield positioning e Dispensers and 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 After start up 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 stabilization of the incubator can take up to 10 minutes The complete stabilization of the electronics will take about one hour so for the best possible performance the instrument should be allowed to stay continuously on for at least one hour It is further recommended to carry out an empty run to verify proper instrument operation The instrument also performs automatic signal long time stability checks during runtime 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 fa
104. tegration time pairs in a SET 8 command are measurement is selected equal 83 LAT module aperture plate position is incorrect Contact service 84 LAT module reference LED failure Contact service 85 Dispensing tip is in wrong position When priming or washing a dispenser the dispensing tip must not be inserted into any dispensing position When dispensing or carrying out tip priming the dispensing tip must be inserted into one of the dispensing positions 86 The command cannot be executed for the wrong plate Use a suitable plate type DME and DIS type commands only work with plate types having up to 384 wells 87 A dispenser has run out of liquid Add liquid and prime the dispenser 88 LAT module base is installed but the LAT module itself is Install the LAT module or replace the base not with a dual dispensing base or a dummy 89 LAT module position calibration failed 90 LAT module position calibration failed 91 TRF lamp fail LAT module flash lamp failure 92 Varioskan LUX LED fail LAT module Varioskan LUX LED failure 93 Integrated gas module is not installed Gas control feature not available 94 Integrated gas module communication error Errors detected in communication with the Integrated gas module 95 Gas concentrations too high 96 Integrated gas module memory error Integrated gas module memory access failed or data was corrupted 97 CO concentration above target Target CO concentration above target 98 CO
105. ters ics 41 Chapter 4 Operations ocio A ri a aaa 43 Operational Chek tree cs A 43 Loading the microplate o ooooooocoocoor RI 44 Automatic runtime calibration RI 46 Measurements and scanning 0 cc cece cette bernia nents 46 FIUCKESCENCEMEASUEMENT serves ice ir E PR nC ie i ks 46 Fluorescence spectrum SCANNING ria a e na a ee eee 49 Absorbance measurement 00 0 t teens 49 ADSOnb ance SPECI UNI Scalig Siria we as Rae aci uc a NN Dai weak Stes 50 Luminescence measurement coco 50 Luminescence spectrum SCANNING n asanare 51 AlphaSereen measuremieril er wei dacs va we A dace MeN 52 oeltl delay AO 52 Measurements under controlled atmosphere 1 sen 53 Other TUFIGUOTIS i d ertet E eria eer e nA pale peras tay elim 53 Track fecha DIIS ETE epe catre nece erectae a c de cec e TR a 53 Orbital Shaka le tc e atc le a ORC ee bea 54 A etre eto e RU Mere a E LENS IMP ICM UE USE IUS 55 DISPONES a du Caos 56 MM a etm ta data A 56 Reagent CONSUMPUON 00d or dida 56 DISPENSING tip oooo oooooooooo RR 57 DISPENSING 2c AO A A Ce ao C et 6 57 IMS SEDET TUTTA 59 MP PMI ss ie a dec d Rc atto rate COPI OU CRDI 60 AAPP ema Ro tb eck cette e 60 Dispensing and measurement isses 60 EMPIINO ti PN atado ata 61 Dispenser WASHING cons Rha CE ACE e C ewan eee 61 Chemical resistance of the diSpenSerS 0 eee eee teens 62 GAS CONTO ci irte anda doen et dada Golder d ick nante id 64 BUTIOSDIIGTB
106. the tip priming vessel either position A or in the front of the tray position B Figure 2 17 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 ul and after 2500 tip primings if the priming volume is 2 ul Pull back feature The pull back feature prevents liquid droplets from forming on the dispensing tip between dispensings Thus the liquid is pulled slightly inwards This function can be set in Skanlt Software or it can be turned off If set the dispenser automatically pulls back 1 to 10 ul 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 Dispensing and measurement 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 44 To minimize the delay before the measurement place the dispensing head into the corresponding dispensing position Refer to Figure 4 44 and Table 4 4 Measurement and dispensing in normal and filter m
107. the transport lock tag under the topmost screw marked number 1 before tightening the screw marked number 2 Tighten the screws marked number 3 and 4 firmly once you have fastened the screws marked number 1 and 2 of the transport lock Figure 5 56 B Replace the front cover by first inserting the two top door latches into place while at the same time turning the cover 30 and then snapping it shut Figure 2 2 and Figure 5 46 Also close the dispenser sliding cover Varioskan LUX 83 Technical Specifications General specifications Thermo Fisher Scientific reserves the right to change any specifications without prior notice as part of our continuous product development program lable 6 6 through Table 6 15 Table 6 6 General specifications General specifications Overall dimensions 526 mm W x 579 mm D x 509 mm H Weight 53 60 kg 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 Performance specification All performance specifications shall be fulfilled within an ambient temperature conditions range of 20 25 C in humidity range of 10 80 Transportation conditions 40 C to 70 C packed in transport packaging Storage conditions 25 C to 50 C packed in transport packaging Mains power supply 100 240 Vac 50 60 Hz
108. tific representative Contacting us For the latest information on products and services visit our websites at http www thermoscientific com http www unitylabservices com Thermo Scientific Table of Contents O E ON 3 About TIS QUIE secet a ad a 3 Related COCUMENTATION vota a dace ic d c ed ol xac IR HR CR RR RR 3 Safety ano Special NOUCES lt lt 4 x ote peperere Re ker ee ped ppc pere E TERRE 3 Instrument safety and guidelines for use iiis 4 Emergeney SItUatiorts serr cea E cese v er eei Y edo Qe ee D e eroe Y 4 Contacting US esos ae pea mor Dre A en opta AUR s 4 Chapter 1 Introduction to Varioskan LUX oooococcccccccc m X eH 9 OQVEIVIEW cercas dad Gor Sa deat dda Gi bathe bbs de dod dtd pedet dete dtd 9 A Dvvcc Im 11 Chapter2 Functional Description 0000 ccc cece eee eee nnn RIR 13 Mstirument AVOUT cet atc a DEN Pd e UD FR eRe eaten ae 13 SONS PM 13 BACK VIEW Si doter Geil den ed ete o asa om v e doeet e 14 INEM VIGW scis Peete bro ooh rd lle ada eu 15 Optical SUS rasp erac dd 16 Principle of the optical system 2 cere eee teen tne ens 16 Excitation OPTICS i giii aa IRR 17 Measurement ODNGS esnie octets atm o oda ate atea a Sa ee aes ea 19 Emission reading module 19 Absorbance measurement module e 20 Fluorescence bottom reading s e hrair aa sannana piraan e 20 LAT measurement mogules copre RE RE en EENT RES 21 Control DUO os ces ec tn ate ect c nt n C RD RR ICA
109. tion sensor Connect the male connector of the dispensing head position sensor to the corresponding female connector on the instrument 3 Insert the dispensing heads into their respective dispensing head holder slots on the left hand side Dispenser 1 and Dispenser 2 of the dispenser 4 Remove the protective cap which protects the thin dispensing tip Figure 3 29 a Figure 3 29 Protective cap removed from P the dispensing tip 0 40 mm 36 Varioskan LUX Thermo Scientific Installation Chapter 3 Figure 3 30 Dispenser assembly Male connector of the dispensing head position sensor Dispensing tube assembly Dispenser syringe and plunger wh 4 Plunger lock screw 5 Valve 6 Aspirate tube assembly 7 Dispensing head 8 Dispensing tip IMPORTANT If the dispensers are not properly installed leakage may occur How to set up the Integrated gas module The optional Integrated gas module is integrated in the instrument You just need to attach the supplied tubes Figure 4 31 between the instrument and a gas supply system The gases required for the module CO and or N are supplied to the device from a separate gas supply system either from gas cylinders or from a central pressurized gas container The layout of the gas supply system must ensure that the operating pressure of the gas supply lines can be set to a range between 0 8 bar min to 1 bar max and that the pressure c
110. tuted 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 aliquots 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 Pipetting instructions Pipette 15 ul of ATP Standard solution and 15 ul 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 Thermo Scientific Thermo Scientific Glossary A absorbance optical density A logarithmic function of the transmission o
111. ure it is level It may be a tight fit A click sound indicates a successful installation Note The adapter will not go to the bottom of the tray if the positioning lever is in the way 6 Push the plate tray into the instrument 7 Replace the front cover by inserting the two top door latches into place while turning the cover 30 and then snapping it shut How to set up the dispensers The two optional dispensers are factory installed Figure 3 29 Although they are factory installed you have to install the dispensing tube assembly manually The complete dispensing tube assembly is packed with the accessories eclee f 9 Figure 3 28 Varioskan LUX with the dispenser sliding cover open j d 1 Reagent bottles 2 Dispensers Thermo Scientific Varioskan LUX 35 Chapter 3 Installation Note that the aspirate tubing Figure 3 30 is factory installed into the right Dispenser 1 and 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 To set up the dispensers 1 Fit the complete dispensing tube assembly Figure 3 30 into the left hole Dispenser 1 and Dispenser 2 of the valve and tighten it finger tight The dispensing tube is used to dispense reagent from the syringe into a microplate 2 Each dispenser is equipped with an individual electronic dispensing head posi
112. urement optics Photometer module Fluorescence bottom reading Fluorescence intensity bottom reading is based on fiber optics that transmits the light from the spectral scanning module to the fluorescence bottom reading optics Figure 2 12 Excitation light Reference sensor measurement optics EX fiber link EM fiber link Thermo Scientific Figure 2 13LAT measurement module Thermo Scientific Functional Description Chapter 2 LAT measurement module The LAT module is an option that is mounted in the front of the unit to the left of the dispensers For an overview of the module see Figure 2 13 LAT module Photo multiplier tube Shutter amp filter selector Measure ment optics Excitation Reference optics system Automatic cross talk shield 96 384 1536 AlphaScreen or Time resolved fluorescence excitation Luminescence AlphaScreen or Time resolved fluorescence emission The LAT 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 correct 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 positio
113. ute period remove the aspirate tubing Figure 3 30 from the 0 1 M HCI solution and remove all the fluid from the syringe and tubing into a waste container 5 Prime the dispenser a minimum of 10 cycles with distilled or deionized water Periodic maintenance 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 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 53 and or the dispensing tip Figure 5 51 2 syringe Figure 5 53 and 3 valve Figure 5 53 If the plunger is stuck Improper washing of a syringe may cause the plunger to get jammed The following may help 1 Remove the syringe see Replacing a dispenser syringe on page 76 and soak it in alcohol or detergent solution 2 Ifthe plunger does not move after this you will need to replace it 3 If the 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
114. vo tica era DRE Recto o oe CR Pa wt we 64 Pom EU 64 A ri eesi trens EDU rc ced eoe e Rap Gee rece A 65 Wd EU 65 Chapter 5 Maintenance svi cece cuoi a nmn ee ele ae a nmm eee nm nn 67 Preventive maintenance 6 Hm 67 How to clean the measurement chamber 68 How to clean the optics in the measurement chamber oooo ooocooooo o 69 How to clean the tray ce ttre m 70 How to clean the reagent basin and dispensing area n isis eee 70 Thermo Scientific Thermo Scientific Table of Contents How to clean the LAT Module osissa assure rris Hn 70 Routine maintenance of the diSPENSerS 6 tet o 72 Daily miallTeralle i ciui aea di c CAE c C n 73 Weekly maihteriarce e iiie reg Force EP e eot oet 73 Weak detergent or 10 bleach n n isses 13 Weak base and acid in SEQUENCE cece eee eee eens 73 Periodic maintenan iesse nennen ce m 74 If the plunger is stuck i e 74 Replacing the aspirate tube assembly or the complete dispensing tube assembly 75 Replacing a dispensing AO x eserse ie rper bw rara A 75 Replacing a dispenser Syringe n n nananana 76 Replacing tfie 3 port ValVe i ox dates cod kai a trees Moc ee Kee un 78 Disposal information se oiai ae a aaa RR E 79 Disposalot matena 5 5 creo rat a e tee cae adus E via 79 Disposal of the instrument isse teen e ene ns 79 Decontamination procedure Hm 80 OW 10 PACK Tor SEIS ue ue t ucc ee oe io Ree Ee cus eec ee d 81 How to refit t

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