tecan Infinite 200 PRO Instructions For Use Manual Download Page 138

Page: 138 / 192

6. Instrument Features

138

IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6

2016-04

6.3

Fluorescence Intensity and
Time Resolved Fluorescence (TRF)

Parameters

Characteristics

Wavelength Range -
Infinite M200 PRO:

230 - 850 nm

selectable in 1 nm steps

Wavelength Range -
Infinite F200 PRO:

230 - 850 nm

Standard Filter:

Not applicable –
instruments are equipped with user-defined filters

Parameters

Instrument type

Characteristics

Wavelength Range

standard

Excitation: 230 – 600 nm

spectrally-
enhanced

Excitation: 230 – 850 nm

standard

Emission: 330 – 600 nm

spectrally-
enhanced

Emission: 280 – 850 nm

Gain Setting

Values

Measurement Range

Manual

1 - 255

0 - 60,000 RFU

Optimal

automatic

0 - 60,000 RFU

Calculated from
Well

automatic

0 - 60,000 RFU

TRF Parameters

Characteristics

Integration Time

10 - 2000 µs

Lag Time

0 - 2000 µs

Summary of Contents for Infinite 200 PRO

Page 1: ...Instructions for Use for Infinite 200 PRO Document Part No 30052730 2016 04 Document Revision No 1 6...

Page 2: ...ts on this publication Manufacturer Tecan Austria GmbH Untersbergstr 1A A 5082 Gr dig Austria T 43 62 46 89 33 F 43 62 46 72 770 E mail office austria tecan com www tecan com Copyright Information The...

Page 3: ...Dual Luciferase and Enliten are registered trademarks of Promega Corporation Madison WI USA Starna is a registered trademark of Starna Scientific Limited 52 54 Fowler Road Hainault Essex IG6 3UT Engla...

Page 4: ...ON NEGATIVE ENVIRONMENTAL IMPACTS ASSOCIATED WITH THE TREATMENT OF WASTE DO NOT TREAT ELECTRICAL AND ELECTRONIC EQUIPMENT AS UNSORTED MUNICIPAL WASTE COLLECT WASTE ELECTRICAL AND ELECTRONIC EQUIPMENT...

Page 5: ...2 4 2 Absorbance 22 2 4 3 Luminescence 23 2 4 4 AlphaScreen AlphaLISA 23 2 5 Optical System 24 2 5 1 Fluorescence Intensity System Infinite M200 PRO 24 2 5 2 Fluorescence Intensity System Infinite F2...

Page 6: ...76 5 3 General Options 76 5 4 Defining Filter Slides Infinite F200 PRO only 78 5 4 1 About Filters 78 5 4 2 Filter Slide and Filter Orientation 78 5 4 3 Installing a Custom Filter 80 5 4 4 Defining t...

Page 7: ...ng Measurements 123 5 13 Cuvette Measurements 124 5 13 1 Cuvette Strip 124 5 13 2 Cuvette Movements 124 5 13 3 i control Cuvette Examples 125 5 14 i control Examples 129 5 15 Finishing a Measurement S...

Page 8: ...Polarization Infinite F200 PRO only 167 7 3 5 Glow Luminescence 169 7 3 6 Absorbance Accuracy 170 7 3 7 Absorbance Wavelength Accuracy 170 7 3 8 Absorbance Baseline Flatness Infinite M200 PRO 171 7 3...

Page 9: ...PLATE DEFINITION FILES THAT ARE RECEIVED WITH THE INSTRUMENT SOFTWARE WE HAVE TAKEN EVERY PRECAUTION TO ENSURE THAT THE PLATE HEIGHTS AND WELL DEPTHS ARE CORRECT ACCORDING TO THE DEFINED PLATE TYPE T...

Page 10: ...ws and guidelines 1 National industrial protection law 2 Accident prevention regulations 3 Safety data sheets of the reagent manufacturers WARNING DEPENDING ON THE APPLICATIONS PARTS OF THE INFINITE 2...

Page 11: ...s conforming to the ANSI SBS standards see 6 13 2 Recommended Types of Microplates for further details Note System Validation by Operating Authority is required The Infinite 200 PRO has been validated...

Page 12: ...nce Intensity FI Bottom Time Resolved Fluorescence TRF Fluorescence Resonance Energy Transfer FRET Time Resolved Fluorescence Resonance Energy Transfer TR FRET Flash Fluorescence with injectors Fluore...

Page 13: ...can be recorded When running a spectrum there is no restriction due to cut off filters The Infinite F200 PRO offers high flexibility for the customization of fluorescence and absorbance measurements s...

Page 14: ...RoHS Orange Logo 5 Label Technical Inspection Agency T V 6 Power Supply Fan 7 Name Plate 8 Label Options Configuration 9 Injector Connection 10 USB Connection 11 Warranty Label ATTENTION REMOVING OR...

Page 15: ...ocal Tecan representative 2 3 Injectors Optional The Infinite 200 PRO can be optionally equipped with an injector module consisting of one or two syringe pumps XE 1000 Tecan Systems located in a separ...

Page 16: ...e Infinite 200 PRO can be equipped with one pump pump A or two pumps pumps A and B One Injector Option one pump An Infinite 200 PRO equipped with one pump allows injections in any SBS conform micropla...

Page 17: ...One 125 ml bottles and two 15 ml bottles for the Two Injectors option two pumps The injector option includes up to two bottle holders that are designed for tubes of different sizes and volumes The bo...

Page 18: ...ftly into the injector port until you hear a clicking noise The instrument contains an injector sensor that checks that the position of the injector carrier for the actions inject and dispense is corr...

Page 19: ...ure 7 Inserting the injector carrier into the injector port CAUTION IF THE INJECTOR CARRIER IS NOT INSERTED CORRECTLY IN THE INJECTOR PORT THE INJECTOR SENSOR WILL NOT DETECT THE INSERTED INJECTOR AND...

Page 20: ...ents Fluorescent molecules emit light of specific wavelength when struck by light of shorter wavelength Stokes Shift In particular a single fluorescent molecule can contribute one fluorescence photon...

Page 21: ...pplications by Brian A Pollok and Roger Heim trends in Cell Biology Vol 9 February 1999 Overview is given in the Review Article Application of Fluorescence Resonance Energy Transfer in the Clinical La...

Page 22: ...slow rotation of the labeled molecule indicating that binding probably did occur A resultant low mP value denotes a fast rotation of a molecule indicating that binding probably did not occur The FP me...

Page 23: ...ween fusion proteins containing Renilla luciferase Rluc and a mutant of the Green Fluorescent Protein GFP The BRETsignal is generated by the oxidation of p a DeepBlueC a coelenterazine derivative that...

Page 24: ...double monochromator 3 Fluorescence top optics 4 Emission double monochromator 5 and fluorescence detection The solid arrows indicate the light path of the excitation light the dashed arrows indicate...

Page 25: ...n 2016 04 IFU for Infinite 200 PRO No 30052730 Rev No 1 6 25 Fluorescence Intensity Bottom Diagram Figure 9 Optical System Fluorescence Bottom Figure 10 Detailed view of excitation and emission double...

Page 26: ...e 3 Many applications only one kind of lamp 4 No warm up time required Condenser Condenser type optics from fused silica focus the flash light onto the entrance slit of the excitation monochromator Fi...

Page 27: ...when the monochromator is set for light with a wavelength of 500 nm and the detector detects a signal of 10 000 counts light with different wavelengths creates a signal of only 10 counts For applicati...

Page 28: ...ber bundle The objective lenses are made from fused silica This material provides high UV transmission and is virtually void of auto fluorescence Excitation Spot Size The size of the fiber bundle cros...

Page 29: ...of excitation light and nonspecific fluorescence The wavelength range is selectable from 330 600 nm in the standard instrument and from 280 850 nm in the spectrally enhanced instrument The bandwidth...

Page 30: ...em of the Infinite F200 PRO instrument 1 Light Source 2 Fluorescence Optics 3 Fluorescence Detection System The fluorescence top system is shown in Figure 11 the bottom system in Figure 12 The solid a...

Page 31: ...2 General Description 2016 04 IFU for Infinite 200 PRO No 30052730 Rev No 1 6 31 Fluorescence Intensity Bottom Diagram Figure 12 Fluorescence intensity bottom system of the Infinite F200 PRO...

Page 32: ...em Excitation Filter Wavelength specific bandpass filters serve to select the wavelength range of interest from the whole spectrum of excitation light coming from the flash lamp Filters are installed...

Page 33: ...excitation and emission wavelengths Mirror Type Reflection Excitation Transmission Emission 50 mirror 230 900 nm 230 900 nm 510 dichroic e g fluorescein 320 500 nm 520 780 nm 650 dichroic AlphaScreen...

Page 34: ...etector A photomultiplier tube PMT is used for the detection of such low light levels as involved with fluorescence For details see section Fluorescence Intensity Detection page 29 2 5 3 Fluorescence...

Page 35: ...is located underneath the plate carrier It measures the light being transmitted through the sample Before measurement of the microplate a reference measurement is performed with the plate carrier mov...

Page 36: ...sed through the dispensed liquid is slightly refracted at the interfaces between air liquid and plate bottom To accomplish a reliable measurement in the presence of the meniscus a focusing lens recoll...

Page 37: ...slit to the excitation monochromator A fiber bundle then guides the light from the excitation monochromator to the absorbance MTP optics which focuses the light into the wells The absorbance MTP meas...

Page 38: ...is 0 7 mm in diameter Absorbance Detection MTP A silicon photodiode is used for the measurement of the transmitted light It is sensitive to a wide range of wavelengths The photodiode is well suited fo...

Page 39: ...t PMT passing through a filter wheel The photon counting PMT photomultiplier tube is designed for applications in chemo and bioluminescence providing a high dynamic range The exceptionally low noise a...

Page 40: ...stem makes it necessary to attenuate high luminescence light levels therefore the filter wheel can also switch a neutral density filter across the selected fiber exit Filter Wheel Position Filter Posi...

Page 41: ...trum of filter Lumi Green Figure 18 Transmission spectrum of filter Blue 1 Figure 19 Transmission spectrum of filter Green 1 0 10 20 30 40 50 60 70 80 90 100 400 450 500 550 600 650 700 750 Wavelength...

Page 42: ...slit to the excitation monochromator A fiber bundle then guides the light from the excitation monochromator to the absorbance cuvette optics which focuses the light through the cuvette The absorbance...

Page 43: ...A silicon photodiode is used for the measurement of the transmitted light It is sensitive to a wide range of wavelengths The photodiode is well suited for the light levels being encountered with absor...

Page 44: ...E M200 PRO CANNOT BE USED FOR CUVETTES WITH A MEASUREMENT WINDOW 2 MM DIAMETER AND A CENTER HEIGHT BELOW 15 MM Inserting the Cuvette The cuvette holder is attached securely to the cuvette carrier and...

Page 45: ...chroic mirror optimized for the AlphaScreen and AlphaLISA wavelengths serves to maximize the excitation and emission light intensities and is selected automatically for Alpha measurements Note AlphaSc...

Page 46: ...measurements Note The dichroic mirror used for AlphaScreen AlphaLISA cannot be used for standard fluorescence measurements It is selected automatically for Alpha measurements PMT Detector AlphaScreen...

Page 47: ...kaging includes additionally the following items Accessory Box Filter stop rings 8 Filter assembly tool Plastic tweezers Filter slide The injector module packaging for 1 injector includes the followin...

Page 48: ...instrument for loose bent or broken parts Report any damage immediately 6 Compare the serial number on the rear panel of the instrument with the serial number on the packing slip Report any discrepan...

Page 49: ...t be damaged Before the instrument can be used the transport lock must be removed using the following procedure 1 Ensure that the instrument is disconnected from the main power supply 2 Open the plate...

Page 50: ...the original packing and installed transport locks Before shipping the instrument it must be thoroughly disinfected see 8 4 Instrument Disinfection 3 3 2 Storage Before storing the instrument the inje...

Page 51: ...rument is correct to this specification The voltage range is 100 120 220 240V If the voltage is not correct please contact your distributor CAUTION DO NOT USE THE INSTRUMENT IF THE VOLTAGE SETTING IS...

Page 52: ...he OFF position 2 Connect the computer to the instrument with the delivered USB interface cable 3 Insert the power cable into the main power socket with protective ground connection on the back panel...

Page 53: ...ors due to their vocational experience are familiar with the necessary safety precautions for handling gas and biohazardous substances Adhere to the following guidelines WARNING The gas control module...

Page 54: ...n can be set in the range from 0 10 vol The CO2 control range is 0 10vol The Accuracy of measured O2 concentration is 0 9 vol The Accuracy of measured CO2 concentration is 1 vol The GCM is available i...

Page 55: ...O2 configuration Carbon Dioxide Mode CO2 Mode Manual Mode o For instruments with CO2 configuration O2 NO SENSOR will appear on the display For the GCM with CO2 configuration O2 Mode and Dual Mode menu...

Page 56: ...30052730 Rev No 1 6 2016 04 Control Panel The control panel which consists of a display panel and a key pad is located on the top of the GCM enhanced 1 Display panel 2 Power gas flow indicator LED 3...

Page 57: ...switch modes 4 3 1 CO2 Mode In CO2 Mode it is possible to regulate CO2 gas concentration by setting the target value and starting the gas control module CO2 gas is introduced into the instrument 1 Sel...

Page 58: ...rument to below ambient air levels 1 Select the mode by pressing the I O key the following display appears 2 Change target value for O2 using the plus and minus keys The editable target value blinks h...

Page 59: ...appears 2 Change target value for CO2 using the plus and minus keys The editable target value blinks highlighted in gray above 3 Press the Mode key to switch to O2 target settings 4 Change target valu...

Page 60: ...display appears 2 Press the Mode key to switch between CO2 and O2 The selected gas will blink highlighted in gray above For instruments with CO2 configuration O2 NO SENSOR will appear 3 Press the I O...

Page 61: ...gs in Device name and Altitude Device name select a letter A B C D for device name using the plus and minus keys when more than one Infinite 200 PRO with GCM is connected to a PC maximum 4 instruments...

Page 62: ...ocate the gas control module near direct sunlight or heat sources Maintain a low dust environment Keep liquids and vapors away from the gas control module Adequate ventilation must be provided for the...

Page 63: ...D SAFETY REGULATIONS WHEN SETTING UP THE CO2 AND OR N2 SUPPLY READ ALL LABEL INFORMATION AND MATERIAL SAFETY DATA SHEETS MSDS FROM MANUFACTURER OR SUPPLIER IMPORTANT ALWAYS USE A REGULATOR APPROVED FO...

Page 64: ...regulator of the cylinder with a plastic clamp as depicted in Figure 26 below Figure 26 3 If the GCM is configured for CO2 and O2 nitrogen gas can be used to regulate the amount of oxygen in addition...

Page 65: ...Infinite 200 PRO using the provided short tube with quick connectors on both ends see Figure 28 below Figure 28 5 Use the provided data cable to connect the module with the Infinite 200 PRO instrument...

Page 66: ...ruptable power outlet 100 240 V 50 60 Hz A green light on the power adapter indicates that it is working see Figure 30 below Figure 30 Power Plug 12 V Power cable for gas control module 7 Connect the...

Page 67: ...quired mode After switching ON the instrument and before switching on gas flow regulation or when valves are closed in Manual Mode the Power gas flow indicator light is on and stable 4 Allow the senso...

Page 68: ...tly and that the valves of the gas cylinder are open to allow the flow of gas Slowly open the main valve of the cylinder or manifold Step 1 then open the shut off valve Step 2 When using CO2 or N2 use...

Page 69: ...FOIL TAPE OR COVER TO THE MICROPLATE SEALING THE PLATE ALLOWS FOR VENTING CULTURES WHILE SIMULTANEOUSLY ACTING AS A BARRIER TO REDUCE EVAPORATION DURING GAS SUPPLY WARNING TREAT BIO HAZARDOUS MATERIA...

Page 70: ...gas cylinder or laboratory gas handling system remove the tubes from the GCM and let out any remaining gas from the tubing Data logging The i control software supports data logging and data display fo...

Page 71: ...bar Make sure that the pressure reduction valve is designed for use with biological applications ask manufacturer The connection from gas cylinder to pressure reduction valve is different for each cou...

Page 72: ...actual current gas concentration is already beyond the target value Check that the target value is not higher than the actual value for O2 Check that the target value is not lower than the actual valu...

Page 73: ...t until actual current gas concentration is below 15 vol e g open plate carrier compartment to speed the reduction of CO2 value before resuming Data cable not connected to the gas control module and o...

Page 74: ......

Page 75: ...tion CAUTION WHEN PLACING A MICROPLATE INTO THE PLATE CARRIER ALWAYS MAKE SURE THAT THE CORRECT PLATE DEFINITION FILE PLATE HEIGHT HAS BEEN SELECTED IN THE SOFTWARE BEFORE YOU DO ANYTHING ELSE MAXIMUM...

Page 76: ...ablished between the instrument and the user interface The following steps are performed 1 OS filter wheels are initialized M200 only 2 Luminescence filter wheel is initialized 3 Z transport of lumine...

Page 77: ...iscus will shift to a lower position giving rise to slightly out of focus conditions Usually wells in the corner evaporate faster the next at the edges of the microplate When measuring fluorescence de...

Page 78: ...commonly used in microplate readers for absorbance measurements usually have a bandwidth of 10 nm Therefore it is not recommended to use fluorescence filters for absorbance measurements because the ba...

Page 79: ...ORE INSERTING A NEW FILTER CAREFULLY CONSIDER THE FILTER AND THE DIRECTION OF LIGHT THROUGH THE FILTER SLIDE 1 Filters with an arrow on the side Light must travel in the direction of the arrow 2 Filte...

Page 80: ...n and emission filters which are placed together with the polarizers either on the position 1 and 2 or 3 and 4 Figure 38 Infinite F200 PRO Filter slide with the indicated positions for fluorescence po...

Page 81: ...olarizer must be inserted into the slide as shown below Note Make sure that the filters are inserted correctly see Filter Types To ensure proper function do not reuse the stop rings more than 5 times...

Page 82: ...slide using tweezers taking care not to scratch it or get fingerprints on it 1 Carefully insert the filter into the opening taking care not to scratch or get fingerprints on the filter 2 Place the sto...

Page 83: ...new undefined customized filter slide is to be used the filter slides need to be defined Depending on the frequency of use and environmental conditions optical filters may deteriorate over time and t...

Page 84: ...LLY ENTERED AVOID USING THE SAME DESCRIPTION FOR THE DIFFERENT FILTER SLIDES Position 1 4 Filter definition editor for the filters filter pairs on positions 1 2 3 and 4 Select the appropriate filter p...

Page 85: ...Wavelength Enter the filter wavelength within the following range 1 Fluorescence intensity mode 230 to 850 nm Excitation and 280 to 850 nm Emission 2 Fluorescence polarization 300 to 850 nm Excitation...

Page 86: ...s saved together with other information about the filter on the filter slide microchip If you replace a filter this information will be lost unless the last filter flash number is manually documented...

Page 87: ...readings of lower concentration microplate wells separate from background as far as the background noise level allows for that Note If any well of interest is assigned OVER overflow you may manually r...

Page 88: ...on can be determined as follows 1 Manual When using the option manual a numeric z position value can be entered in the measurement stripe The default manual z position is 20000 m 2 Calculated from wel...

Page 89: ...ition optimization shall be performed The optimal z position can be simultaneously determined for up to 4 labels The label selection number of labels depends on the measurement script previously defin...

Page 90: ...imization Select the well s and click Scan to start the z optimization The z positioning option Max S B Ratio requires the measurement of two wells one filled with a fluorophore of interest signal and...

Page 91: ...is then applied to the second measurement with the blank well Therefore both signal and blank curves are directly comparable The z position for each selected label can be defined manually In the grap...

Page 92: ...al Integration Time is delayed against the preceding flash by a Lag Time TRF timing parameters may be established with the following procedure 1 As a starting point you may take the Fluorescence Lifet...

Page 93: ...e M200 PRO can switch between two wavelengths within 150 ms in case that the selected labels are measured with the same gain and no order sorting OS switching point is involved see Table 1 for switchi...

Page 94: ...e filled 21 24 well 12 well Circle filled 37 12 well 6 well Circle filled 76 6 well Table 2 Optimal Read spot patterns in different plate formats Changing the total number of flashes per well 1 100 wi...

Page 95: ...flashes as well as the number of flashes per well are also displayed Figure 39 Results output for a measurement with Optimal Read example for a 96 well plate Miscellaneous Features of Optimal Read Opt...

Page 96: ...tion is a dimensionless unit generally expressed in mP units To start an FP measurement the program strip must contain a valid measurement Blank range and valid G Factor settings 5 6 2 Measurement Bla...

Page 97: ...h of the fluorophore by measuring a sample with a known polarization value A valid calibration of the instrument resulting in a G factor is an important requirement for each fluorescence polarization...

Page 98: ...ce value select a polarization value for the reference used e g 20 mP for a 1 nM Fluorescein solution in 0 01 M NaOH Reference range click Change and select the wells filled with the reference Blank r...

Page 99: ...ter slides and thus stored with the different filter slide descriptions Caution The filter slide description is part of the G Factor key value Avoid using the same filter slide description for differe...

Page 100: ...he displayed G Factor does not match the calibrated value e g the G Factor has been manually changed or loaded with a method the corresponding value will be marked as Manual G Factor The calibrated G...

Page 101: ...uction The mean value of the respective blank wells is subtracted from each value well each for RFU RFU RFU RFU RFU RFU RFU RFU RFU par blk par blk par blk par smp par buf par buf par buf par ref par...

Page 102: ...rise to fluctuations of the measured values The effect has been observed in wells of 96 well plates and larger wells 5 7 2 Absorbance Ratio Mode Ratio Mode Using the Standard tab in i control up to 4...

Page 103: ...ogram strip by selecting the Multiple Reads per Well check box see Figure 40below Figure 40 Multiple Reads per Well Note The function Multiple Reads per Well is only available for the fixed wavelength...

Page 104: ...e optics 0 7 mm 0 5 mm Table 3 Theoretically calculated beam diameter at the focal point The MRW type displayed in the software is therefore only a schematic overview of the measurement pattern When m...

Page 105: ...m and wall of the microplate well CAUTION ALL ABSORBANCE AND FLUORESCENCE INTENSITY SPECIFICATIONS GIVEN IN THIS DOCUMENT ARE ONLY VALID FOR SINGLE POINT MEASUREMENTS ONE MEASUREMENT POINT PER WELL WH...

Page 106: ...41 Alignment graphic XY Line 3 x 3 for result of a fluorescence measurement In addition the standard deviation Stdev and the average value Mean of the measurements points well are also displayed Figu...

Page 107: ...yields more accurate values because of the irregular photon impact photon statistics The photonic noise shot noise cannot be reduced technically Note The signal to noise S N ratio can be optimized by...

Page 108: ...rovide a proper signal to noise ratio S N on the one hand and linearity on the other hand Therefore the gain should be tuned to make highest concentration microplate wells give highest possible readin...

Page 109: ...he intended use of the Infinite 200 PRO PMT is therefore specified for gain settings from 60 to 255 Gain settings below 60 are possible but the performance of the PMT is not specified for voltage supp...

Page 110: ...the holder of the injector carrier system For priming and washing steps of the injector system a default setting for injection speed and volume dispensed is provided If required the priming parameters...

Page 111: ...ming Procedure i control 1 Adjust parameters on the Prime tab of the Injector Maintenance dialog box in the Settings menu 2 Start the priming procedure by clicking Start prime in the Injector Maintena...

Page 112: ...ement with the actions injection or dispense 5 l of liquid are dispensed into a disposable container on the plate carrier before starting injection or dispense This initial dispense step makes sure th...

Page 113: ...Injector B or Injector A and B only primed injectors are available for backflush 12 Select the Piston Strokes 1 60 1 stroke equals 1 ml 13 Select the Backflush Speed 100 300 l sec 14 Select the Refill...

Page 114: ...e tubes reach the bottom of the bottle 2 Remove the injector from the carrier slot and insert it into the service position of the injector box 3 Put an empty container under the injector Washing Proce...

Page 115: ...E INJECTOR CARRIER MUST BE IN THE SERVICE POSITION FOR THE ACTION WASH DO NOT PERFORM WASHING WHEN THE INJECTOR IS IN THE INSTRUMENT IMPORTANT BE SURE TO RUN A FINAL WASH PROCEDURE WITH DISTILLED WATE...

Page 116: ...se step depends on the selected refill mode selected on the injector or dispense strip see chapter 5 10 5 Injector for details When using standard refill mode the dispense step is performed after each...

Page 117: ...CAUTION PLACE THE WASTE TUB ON THE PLATE TRANSPORT BEFORE STARTING A MEASUREMENT WITH THE ACTIONS INJECTION AND OR DISPENSE CAUTION IT IS RECOMMENDED TO EMPTY THE WASTE TUB BEFORE STARTING A MEASUREME...

Page 118: ...placement for any reason after the tubes have been changed do not forget to perform washing and priming before starting a measurement 5 11 4 Injector Modes i control When using the injector two modes...

Page 119: ...strip does not require an additional measurement strip Dispense volume The injection volume depends on the microplate type The plate definition files include a so called working volume This working v...

Page 120: ...efill mode Standard if refill should be performed when syringe is empty multiple injection steps are performed before refilling refill occurs after dispensing approx 800 l Select Refill for every inje...

Page 121: ...ing indented strips are performed well wise Injection Set up the injection parameters Only one injector can be selected per strip If both injectors are required or one injector will perform two inject...

Page 122: ...tion file is higher than the volume used for injection Wait Strip A Wait time delay or settle time can be inserted into the procedure Wait time Select a time in hh mm ss from 00 00 01 up to 23 59 59 O...

Page 123: ...spreadsheet The data are also stored in the software and can be applied to the following cuvette measurements performed with the same parameters The blank data are automatically subtracted when the c...

Page 124: ...one cuvette strip and one plate strip within one measurement script The cuvette measurement has to be positioned before the microplate measurement To perform an accurate microplate measurement the cu...

Page 125: ...2730 Rev No 1 6 125 5 13 3 i control Cuvette Examples Example 1 Example of how to use the Blanking measurement when measuring a DNA sample Prepare cuvette with sample buffer Set up the DNA measurement...

Page 126: ...blank cuvette Prepare sample cuvette and put it on the cuvette holder Start the measurement by clicking Start The cuvette holder is moved in and the measurement is performed The measured data Value a...

Page 127: ...fixed wavelength strip Apply blanking is disabled when a reference wavelength is selected Select the appropriate measurement parameters wavelength number of flashes and settle time Plate Necessary for...

Page 128: ...he measurement and therefore allows removing the cuvette from the cuvette port When confirming the request the measurement continues Move Cuvette In The cuvette port is moved in Plate Necessary for mi...

Page 129: ...ell Mandatory for measurements with injection Injection 1 Injector A injects 100 l with speed 200 l sec refill mode standard Wait Timer 2 s wait time Luminescence 1 Luminescence measurement with 10 s...

Page 130: ...ed Part of the plate Not shown can be optionally selected if only part of the plate should be processed Well Mandatory for measurements with injection Injection Injector A injects 100 l with speed 100...

Page 131: ...actions to be performed once in a kinetic run at a certain cycle The intended injection strip below it is only processed once at the selected cycle Injection Injector A injects 20 l with speed 200 l s...

Page 132: ...actions to be performed once in a kinetic run at a certain cycle The intended injection strip below it is only processed once at the selected cycle Injection Injector A injects 20 l with speed 200 l s...

Page 133: ...the instrument and the computer is terminated Note Remove the microplate and or cuvette before disconnecting the instrument from the computer 5 15 2 Instrument Shut Down Upon shut down the instrument...

Page 134: ......

Page 135: ...and Time Resolved Fluorescence TRF Fluorescence Polarization See 6 4 Fluorescence Polarization FP Infinite F200 PRO only Absorbance See 6 5 Absorbance Glow Type Luminescence See 6 6 Glow Type Luminesc...

Page 136: ...ing Linear and orbital shaking amplitude selectable from 1 6 mm in 0 5 mm steps Light Source High energy xenon flash lamp life time 10 8 flashes Optics Fused Silica Lenses Detectors Fluorescence Stand...

Page 137: ...Environmental Ambient Temperature Operation 15 C to 30 C 59 F to 86 F Non operation 20 C to 60 C 4 F to 140 F Relative Humidity Operation 80 non condensing Over voltage Category II Usage General Labo...

Page 138: ...ard Filter Not applicable instruments are equipped with user defined filters Parameters Instrument type Characteristics Wavelength Range standard Excitation 230 600 nm spectrally enhanced Excitation 2...

Page 139: ...Type number of wells 96 Dispensed Volume l 200 Flashes Reads per Well 25 Fluorescein Detection Limit pM 20 pM Uniformity at 25 nM Fluorescein 3 CV Reproducibility at 25 nM Fluorescein 2 6 3 3 Fluoresc...

Page 140: ...anced Excitation 300 850 nm standard Emission 330 600 nm spectrally enhanced Emission 330 850 nm Standard Filter both Configuration of Default filter slide Exc 485 20 nm Position on Exc filter slide E...

Page 141: ...ble Measurement Range Both 0 4 OD The following specifications are valid for the wavelength range from 300 700 nm for Infinite 200 PRO series Plate type number of wells 96 Accuracy 0 2 OD 1 10 mOD Acc...

Page 142: ...cteristics Wavelength Range 380 600 nm Linear Dynamic Range 6 orders of magnitude Integration Time well 100 20000 ms Cross Talk black plate 0 01 Measurement range 6 orders of magnitude 8 orders of mag...

Page 143: ...EMENT Luminescence Detection uses the photon counting technique Parameters Characteristics Wavelength Range 380 600 nm Measurement Range 6 orders of magnitude 8 orders of magnitude extended dynamic ra...

Page 144: ...MEASUREMENT Parameters Characteristics Built in wavelength See 2 5 4 Absorbance System Infinite F200 PRO Integration time 100 20000 ms Different integration times are possible for each wavelength Pla...

Page 145: ...rd Filters AlphaLISA Excitation 680 30 nm Emission 615 20 nm Gain Setting Values Measurement Range Manual 1 255 0 60 000 RFU Calculated from Well automatic 0 60 000 RFU Parameters Characteristics Exci...

Page 146: ...tion in 1 nm steps possible Measurement Range 0 4 OD 6 11 1 Cuvette Specifications The following specifications are valid for the wavelength range from 300 700 nm number of flashes 25 Cuvette types 12...

Page 147: ...ls with a rating D must not be used with the Infinite injectors They will severely damage the injector system A Rated Chemicals D Rated Chemicals Acetic Acid 60 Butyl Amine Acetonytrile Carbon Tetrach...

Page 148: ...aler for a recommended filter set Filters designed for a different type of instrument will not necessarily perform well with the Infinite F200 PRO Note If the excitation and the emission maximum of a...

Page 149: ...from 6 to 384 wells maximum plate height 23 mm including lid that conform to the following standards can be measured ANSI SBS 1 2004 ANSI SBS 2 2004 ANSI SBS 3 2004 and ANSI SBS 4 2004 When installin...

Page 150: ...fb 782 076 782 077 AC 782061 AC 78207x AC 782101 GRE96ft_half area 675 161 675 101 675 801 AC 675801 GRE96fw_half area 675 074 675 075 675 094 675 095 AC 675801 GRE96fb_half area 675 077 675 076 675 0...

Page 151: ...om 3711 DWG00682 COR384fw clear bottom 3706 DWG00682 COR384fc UV transparent 3675 DWG01479 COR1536fb 3724 DWG01840 COR1536fw 3725 DWG01840 COR1536fb clear bottom 3891 DWG01543 Nunclon NUN96ft 439 454...

Page 152: ...OptiPlate 6005290 http www perki nelmer com Cat alog Product ID 6005290 PE96fw_ProxiPlate 6006290 http www perki nelmer com Cat alog Product ID 6006290 PE384fg_AlphaPlate 6008350 TechnicalDataS heet_D...

Page 153: ...technique This is based on a dedicated luminescence PMT with appropriate measurement circuitry This technique is very robust against noise It is preferred for measurement of very low light levels For...

Page 154: ......

Page 155: ...uctions carefully The user should determine the appropriate intervals for this testing based on how frequently the instrument is operated We recommend adapting these tests and the acceptance criteria...

Page 156: ...cence Top Uniformity 3 CV Fluorescence Top Precision 2 CV Fluorescence Bottom Sensitivity Standard 100 pM Enhanced Infinite M200 PRO 45 pM Infinite F200 PRO 35 pM Fluorescence Bottom Uniformity 3 CV F...

Page 157: ...uctions are valid for the Infinite 200 PRO Infinite F200 PRO Infinite M200 PRO Standard version Spectrally enhanced version Sensitivity Perform the following measurement to determine the detection lim...

Page 158: ...agents 1 nM Fluorescein in 0 01 M NaOH Fluorescein sodium salt Sigma F6377 0 01 M NaOH Blank NaOH pellets Merck article no 6495 or Sigma S8045 1 Greiner 96 well plate black 200 l Pipette tips Calculat...

Page 159: ...olution 0 01 M NaOH into the appropriate wells according to the plate layout 1 2 3 4 5 6 7 8 9 10 11 12 A 1 nM Fluorescein Blank 1 nM Fluorescein Blank 1 nM Fluorescein Blank 1 nM Fluorescein Blank 1...

Page 160: ...M Fluorescein or the blank solution 0 01 M NaOH into the appropriate wells according to the plate layout 1 2 3 4 5 6 7 8 9 10 11 12 A 1 nM Fluorescein Blank 1 nM Fluorescein Blank 1 nM Fluorescein Bla...

Page 161: ...ing measurement to determine the detection limit for Fluorescein Measurement Parameters Parameter Setting Reading Mode Fluorescence Bottom Ex Wavelength Infinite F200 PRO 485 20 nm Infinite M200 PRO 4...

Page 162: ...rmine the detection limit for Fluorescein Measurement Parameters Parameter Setting Reading Mode Fluorescence Bottom Ex Wavelength Infinite F200 PRO 485 20 nm Infinite M200 PRO 485 nm Em Wavelength Inf...

Page 163: ...Ex Wavelength Infinite F200 PRO 485 20 nm Infinite M200 PRO 485 nm Em Wavelength Infinite F200 PRO 535 25 nm Infinite M200 PRO 535 nm Number of flashes 25 Integration Time 40 Settle Time 0 Gain Optima...

Page 164: ...e Time 0 Gain Optimal Plate Type GRE96fb Part of the Plate A1 Kinetic 20 Cycles Interval Time Minimal Plate Layout 1 2 3 4 5 6 7 8 9 10 11 12 A 25 nM Fluorescein Blank 25 nM Fluorescein Blank 25 nM Fl...

Page 165: ...measurement to determine the sensitivity Measurement Parameters Parameter Setting Reading Mode Fluorescence Top Ex Wavelength Infinite F200 PRO 340 35 nm Infinite M200 PRO 340 nm Em Wavelength Infinit...

Page 166: ...surement to determine the precision reproducibility Measurement Parameters Parameter Setting Reading Mode Fluorescence Top Ex Wavelength Infinite F200 PRO 340 35 nm Infinite M200 PRO 340 nm Em Wavelen...

Page 167: ...e valid for the Infinite F200 PRO Standard version Spectrally enhanced version Precision Perform the following measurement to determine the precision Measurement Parameters Parameter Setting Reading M...

Page 168: ...Blank B C D E F G H Filling volume 200 l well Material Reagents 1 nM Fluorescein in 0 01 M NaOH Fluorescein sodium salt Sigma F6377 0 01 M NaOH Blank NaOH pellets Merck Article No 6495 or Sigma S8045...

Page 169: ...nce Integration Time 1000 ms Settle Time 0 Plate Type GRE96fw Part of the Plate A1 D10 Plate Layout Pipette 200 l of the ATP reagents into the appropriate wells according to the plate layout 1 2 3 4 5...

Page 170: ...racy Use MultiCheck Plate For details please refer to the MultiCheck Instructions for Use 7 3 7 Absorbance Wavelength Accuracy The wavelength accuracy defines the deviation of the set measurement wave...

Page 171: ...t Parameters Parameter Setting Reading Mode Absorbance Measurement Wavelength from to 300 700 nm Number of flashes 25 Settle Time 0 Plate Type GRE96ft Part of the Plate A1 Plate Layout No plate is nec...

Page 172: ...ing Mode Absorbance Measurement Wavelength 340 nm 405 nm 492 nm 590 nm 620 nm 700 nm Number of flashes 25 Settle Time 0 Plate Type GRE96ft Part of the Plate A1 Kinetic Cycles 20 Minimal Interval Time...

Page 173: ...etermine the absorbance accuracy Measurement Parameters Parameter Setting Reading Mode Absorbance Measurement Wavelength 1 440 nm 2 635 nm Number of flashes 25 Time between Move and Read 0 Cuvette Typ...

Page 174: ...100 l pipette tips Omnibeads dilution series Dilute the Omnibeads stock solution 1 500 in PBS by adding 3 l of the stock solution 5 mg ml to 1497 l PBS yielding a solution of 10 g ml Prepare 12 furth...

Page 175: ...on for each Omnibead concentration Perform a blank reduction by subtracting the average signal of the blanks wells from the average signal of each Omnibead concentration Plot the average blank correct...

Page 176: ......

Page 177: ...he instrument s life and to reduce the need for servicing This section contains the following procedures Liquid Spills Instrument Disinfection Disinfection Certificate Instrument and Material Disposal...

Page 178: ...NG TO AVOID POTENTIAL INFECTIOUS DISEASE CONTAMINATION ADDITIONALLY ALL RESULTING WASTE FROM THE CLEAN UP MUST BE TREATED AS POTENTIALLY INFECTIOUS AND THE DISPOSAL MUST BE PERFORMED ACCORDING TO THE...

Page 179: ...ekly to remove precipitates and eliminate bacterial growth Follow these steps to clean the pump injector system with 70 EtOH ethanol 1 Depending on the user s application flush thoroughly the system w...

Page 180: ...TED AS POTENTIALLY INFECTIOUS AREAS IT IS ADVISABLE TO ADHERE TO APPLICABLE SAFETY PRECAUTIONS INCLUDING THE WEARING OF POWDER FREE GLOVES SAFETY GLASSES AND PROTECTIVE CLOTHING TO AVOID POTENTIAL INF...

Page 181: ...clearly visible See 8 4 3 Safety Certificate for an example of the safety certificate which must be completed before the instrument is returned to the service center for service or repair 8 4 3 Safet...

Page 182: ...ECT WASTE ELECTRICAL AND ELECTRONIC EQUIPMENT SEPARATELY 8 4 5 Disposal of Packing Material According to Directive 94 62 EC on packaging and packaging waste the manufacturer is responsible for the dis...

Page 183: ...ON ALWAYS DISINFECT THE INSTRUMENT BEFORE DISPOSAL Pollution degree 2 IEC EN 61010 1 Method of disposal Contaminated waste WARNING DEPENDING ON THE APPLICATIONS PARTS OF THE HYDROFLEX PLATFORM MAY HAV...

Page 184: ......

Page 185: ...in the Instrument Computer communication protocol Please report this error to your local Tecan customer support office 9 Binary Transfer command cmd at module n Unspecific internal communication erro...

Page 186: ...ernal communication error Please report this error to your local Tecan customer support office 29 Command Overflow Unspecific internal communication error Please report this error to your local Tecan...

Page 187: ...gth lambda Gain g Counts cts Unspecific Hardware failure Please report this error to your local Tecan customer support office 44 Steploss Error Actuator failure Please report this error to your local...

Page 188: ......

Page 189: ...ssion Filter 34 46 Excitation 32 Excitation Spot Size 28 F Filter Switch Time 93 Filter Wheel PMT 29 Filters recommended 148 Finishing a Measurement Session 133 Flash lamp 26 Flash Luminescence 23 Fla...

Page 190: ...al Disposal 182 Returning 182 PMT Properties 87 109 Polarization 101 Power Requirements 51 Precision Bottom Fluorescence 164 Fluorescence Polarization 167 Time resolved fluorescence 166 Top Fluorescen...

Page 191: ...ntact information Prior to contacting Tecan for product support prepare the following information for the best possible technical support see name plate Model name of your product Serial number SN of...

Page 192: ...ation 2014 30 EU EMC Directive 2006 42 EC Machinery Directive and that the standards referenced below were taken in consideration EN 61010 1 2001 AC 2002 Safety requirements for electrical equipment f...

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