tecan Infinite 200 PRO User manual

Instructions for Use for
Infinite 200 PRO
Document Part No.: 30052730
2016-04
Document Revision No.: 1.6

2 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 2016-04
WARNING
CAREFULLY READ AND FOLLOW THE INSTRUCTIONS PROVIDED
IN THIS DOCUMENT BEFORE OPERATING THE INSTRUMENT.
Notice
Every effort has been made to avoid errors in text and diagrams; however, Tecan
Austria GmbH assumes no responsibility for any errors, which may appear in this
publication.
It is the policy of Tecan Austria GmbH to improve products as new techniques
and components become available. Tecan Austria GmbH therefore reserves the
right to change specifications at any time with appropriate validation, verification,
and approvals.
We would appreciate any comments on this publication.
Manufacturer
Tecan Austria GmbH
Untersbergstr. 1A
A-5082 Grödig, Austria
T [phone]
F [phone]
E-mail: [e-mail]
www.tecan.com
Copyright Information
The contents of this document are the property of Tecan Austria GmbH and are
not to be copied, reproduced or transferred to another person or persons without
prior written permission.
Copyright Tecan Austria GmbH
All rights reserved.
Printed in Austria
Declaration for EU Certificate
See the last page of these Instructions for Use.
About the Instructions for Use
Original Instructions. This document describes the Infinite 200 PRO
multifunctional microplate reader. It is intended as reference and instructions for
use. This document instructs how to:
•Install the instrument
•Operate the instrument
•Clean and maintain the instrument

2016-04 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 3
Remarks on Screenshots
The version number displayed in screenshots may not always be the one of the
currently released version. Screenshots are replaced only if content related to
application has changed.
Trademarks
The following product names and any registered and unregistered trademarks
mentioned in this document are used for identification purposes only and remain
the exclusive property of their respective owners:
•Infinite®, i-controlTM, GCMTM, magellanTM, NanoQuant PlateTM,Tecan®
and the Tecan Logo are registered trademarks of
Tecan Group Ltd., Männedorf, Switzerland
•Windows®and Excel®are registered trademarks of Microsoft Corporation,
Redmond, WA, USA
•ChromaGloTM 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 England, United Kingdom
•BRET2TM, DeepBlueC®, PerkinElmer®, AlphaScreen®and AlphaLISA®are
registered trademarks of PerkinElmer, Inc., Waltham, Massachusetts, USA
Warnings, Cautions, and Notes
The following types of notices are used in this publication to highlight important
information or to warn the user of a potentially dangerous situation:
Note
Gives helpful information.
CAUTION
INDICATES A POSSIBILITY OF INSTRUMENT DAMAGE OR DATA LOSS
IF INSTRUCTIONS ARE NOT FOLLOWED.
WARNING
INDICATES THE POSSIBILITY OF SEVERE PERSONAL INJURY,
LOSS OF LIFE OR EQUIPMENT DAMAGE IF THE INSTRUCTIONS
ARE NOT FOLLOWED.
WARNING
THIS SYMBOL INDICATES THE POSSIBLE PRESENCE OF
BIOLOGICALLY HAZARDOUS MATERIAL. PROPER LABORATORY
SAFETY PRECAUTIONS MUST BE OBSERVED.

4 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 2016-04
WARNING
THIS SYMBOL INDICATES THE POSSIBLE PRESENCE OF
FLAMMABLE MATERIALS AND A RISK OF FIRE. PROPER
LABORATORY SAFETY PRECAUTIONS MUST BE OBSERVED.
ATTENTION
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 SEPARATELY.
Symbols
Manufactured by
Date of manufacture
Conformité Européenne
Read the Instructions for Use
before operating the instrument
Order number
Serial Number
USB label
WEEE symbol
RoHS Orange Logo
TÜV NRTL

2016-04 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 5
Table of Contents
1. Safety.................................................................................................................9
1.1 Instrument Safety.................................................................................9
2. General Description........................................................................................11
2.1 Instrument...........................................................................................11
2.1.1 Intended Use........................................................................................11
2.1.2 Multifunctionality...................................................................................12
2.1.3Performance.........................................................................................13
2.1.4 User Friendliness.................................................................................13
2.1.5 Onboard Control Button .......................................................................13
2.1.6 Rear View.............................................................................................14
2.2 Software..............................................................................................15
2.3 Injectors (Optional) ............................................................................15
2.3.1 Injector Measurement Modes...............................................................15
2.3.2 Injector Module Diagram......................................................................16
2.3.3 Injector Pump Options..........................................................................16
2.3.4 Storage Bottles and Bottle Holders......................................................17
2.3.5 Injector Carrier .....................................................................................18
2.4 Measurement Techniques .................................................................20
2.4.1 Fluorescence........................................................................................20
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 F200 PRO).............................30
2.5.3 Fluorescence Polarization System (Infinite F200 PRO).......................34
2.5.4 Absorbance System (Infinite F200 PRO).............................................35
2.5.5 Absorbance System (Infinite M200 PRO).............................................37
2.5.6 Luminescence System.........................................................................39
2.5.7 Cuvette Port (Infinite M200 PRO).........................................................42
2.5.8 AlphaScreen/AlphaLISA System (Infinite F200 PRO only)...................45
3. Installation.......................................................................................................47
3.1 Unpacking and Inspection.................................................................47
3.1.1 Unpacking Procedure...........................................................................48
3.2Removal of the Transport Locks.......................................................49
3.3 Transport and Storage.......................................................................50
3.3.1 Transport..............................................................................................50
3.3.2 Storage.................................................................................................50
3.4 Power Requirements..........................................................................51
3.5 Switching the Instrument On ............................................................52
4. Gas Control Module (Enhanced) ...................................................................53

6 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 2016-04
4.1 Safety...................................................................................................53
4.2 Gas Control Module Features............................................................54
4.2.1 Gas Control Module Configurations......................................................54
4.2.2 Top and Rear Views of the Gas Control Module ..................................55
4.3 Main Menu of Modes ..........................................................................57
4.3.1 CO2 Mode............................................................................................57
4.3.2 O2 Mode...............................................................................................58
4.3.3 DUAL Mode..........................................................................................59
4.3.4 Manual Mode........................................................................................60
4.4 Settings Menu.....................................................................................61
4.5 Installing the Gas Control Module ....................................................62
4.5.1 Requirements.......................................................................................62
4.5.2 Installation Procedure...........................................................................63
4.6 Operating the Gas Control Module...................................................67
4.7 CO2and N2Gas Cylinders (Not Supplied Accessory).....................71
4.8 Troubleshooting the Gas Control Module........................................72
5. Operating the Instrument ...............................................................................75
5.1 Introduction.........................................................................................75
5.2 General Operating Features ..............................................................76
5.2.1 Instrument Start Up ..............................................................................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 the Filters................................................................................83
5.5 Optimizing Fluorescence Measurements.........................................87
5.5.1 Instrument Parameters.........................................................................87
5.5.2 Z-Optimization (FI Top measurements with the Infinite M200 PRO only)88
5.5.3 FI Ratio Mode.......................................................................................93
5.5.4 Optimal Read........................................................................................94
5.6 FP Measurements...............................................................................96
5.6.1 Fluorescence Polarization ....................................................................96
5.6.2 Measurement Blank Range..................................................................96
5.6.3 G-Factor Settings .................................................................................97
5.6.4Measurement with an Uncalibrated G-Factor.......................................97
5.6.5 Measurement with a Simultaneous G-Factor Calibration .....................98
5.6.6 Measurement with a Calibrated G-Factor.............................................99
5.6.7 Measurement with a Manual G-Factor ...............................................100
5.6.8 Calculation of Fluorescence Polarization Parameters........................101
5.7 Optimizing Absorbance Measurements .........................................102
5.7.1 Measurement Parameters..................................................................102

2016-04 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 7
5.7.2 Absorbance Ratio Mode.....................................................................102
5.8 Multiple Reads per Well...................................................................103
5.8.1 MRW Type.........................................................................................103
5.8.2 MRW Size ..........................................................................................104
5.8.3 MRW Border ......................................................................................104
5.8.4 Result Display in MS Excel ................................................................106
5.8.5 Miscellaneous Software Features of MRW ........................................106
5.9 Optimizing Luminescence Measurements.....................................107
5.9.1 Integration Time .................................................................................107
5.9.2 Light Level Attenuation.......................................................................107
5.10 Optimizing AlphaScreen/AlphaLISA Measurements.....................108
5.10.1 Instrument Parameters.......................................................................108
5.11 Measurements with Injectors..........................................................110
5.11.1 Priming and Washing of the Infinite 200 PRO....................................110
5.11.2 Washing.............................................................................................114
5.11.3 Before Starting a Measurement with Injectors....................................118
5.11.4 Injector Modes (i-control)....................................................................118
5.12 Blanking 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 Session..................................................133
5.15.1 Disconnecting the Instrument.............................................................133
5.15.2 Instrument Shut Down........................................................................133
6. Instrument Features .....................................................................................135
6.1 Introduction ......................................................................................135
6.2 Instrument Specifications ...............................................................136
6.3 Fluorescence Intensity and Time Resolved Fluorescence (TRF).138
6.3.1 Definition of the Detection Limit..........................................................139
6.3.2 Fluorescein (Fluorescence Intensity) Top ..........................................139
6.3.3 Fluorescein (Fluorescence Intensity) Bottom.....................................139
6.3.4 Europium (Time Resolved Fluorescence)..........................................139
6.4 Fluorescence Polarization (FP).......................................................140
6.5 Absorbance ......................................................................................141
6.6 Glow Type Luminescence ...............................................................142
6.6.1 ATP Glow Luminescence...................................................................142
6.7 Flash Type Luminescence...............................................................143
6.8 Dual Color Luminescence (e.g. BRET)...........................................144
6.9 AlphaScreen/AlphaLISA..................................................................144
6.10 “On the Fly” Measurements............................................................145

8 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 2016-04
6.11 Cuvette Features (Infinite M200 PRO only).....................................146
6.11.1 Cuvette Specifications........................................................................146
6.12 Injector Specifications .....................................................................147
6.12.1 Injector Reagent Compatibility............................................................147
6.13 Measurement Accessories ..............................................................148
6.13.1 Recommended Filters (Infinite F200 PRO only).................................148
6.13.2 Recommended Types of Microplates.................................................149
6.13.3 Luminescence Detection....................................................................153
7. Quality Control ..............................................................................................155
7.1 Periodic Quality Control Tests........................................................155
7.2 Specifications - Passed/Failed Criteria...........................................156
7.3 Specifications - Test Instructions...................................................157
7.3.1 Fluorescence Top...............................................................................157
7.3.2 Fluorescence Bottom..........................................................................161
7.3.3 Time Resolved Fluorescence.............................................................165
7.3.4 Fluorescence 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
7.3.9 Absorbance Baseline Flatness (Infinite F200 PRO) ...........................172
7.3.10 Absorbance Cuvette (Infinite M200 PRO only)...................................173
7.3.11 AlphaScreen/AlphaLISA (Infinite F200 PRO only)..............................174
8. Cleaning and Maintenance...........................................................................177
8.1 Introduction.......................................................................................177
8.2 Liquid Spills......................................................................................178
8.3 Injector Cleaning and Maintenance ................................................178
8.3.1 Daily Maintenance:.............................................................................179
8.3.2 Weekly/Periodical Maintenance:.........................................................179
8.4 Instrument Disinfection ...................................................................180
8.4.1 Disinfection Solutions.........................................................................180
8.4.2 Disinfection Procedure .......................................................................181
8.4.3 Safety Certificate................................................................................181
8.4.4 Disposal..............................................................................................182
8.4.5 Disposal of Packing Material..............................................................182
8.4.6 Disposal of Operating Material ...........................................................182
8.4.7 Disposal of the Instrument..................................................................183
9. Troubleshooting............................................................................................185
Index.......................................................................................................................189

1. Safety
2016-04 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 9
1. Safety
1.1 Instrument Safety
1. Always follow basic safety precautions when using this product to reduce
the risk of injury, fire, or electrical shock.
2. Read and understand all information in the Instructions for Use. Failure to
read, understand, and follow the instructions in this document may result
in damage to the product, injury to operating personnel or poor
instrument performance.
3. Observe all WARNING and CAUTION statements in this document.
4. Never open the housing of the Infinite 200 PRO while the instrument is
plugged into a power source.
5. Never force a microplate into the instrument.
6. The Infinite 200 PRO is intended as a general purpose laboratory
instrument for professional use. Observe proper laboratory safety
precautions, such as wearing protective clothing and using approved
laboratory safety procedures.
CAUTION
TECAN AUSTRIA GMBH HAS TAKEN GREAT CARE IN CREATING THE
STORED 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. THIS PARAMETER IS USED TO DETERMINE THE MINIMUM
DISTANCE BETWEEN THE TOP OF THE PLATE AND THE CEILING OF THE
MEASUREMENT CHAMBER. ADDITIONALLY, TECAN AUSTRIA HAS ADDED A
VERY SMALL SAFETY GAP TO PREVENT ANY DAMAGE OCCURRING TO THE
MEASUREMENT CHAMBER AS A RESULT OF SMALL CHANGES IN PLATE
HEIGHT. THIS DOES NOT AFFECT THE PERFORMANCE OF THE
INSTRUMENT.
USERS MUST ENSURE THAT THE PLATE DEFINITION FILE SELECTED
CORRESPONDS TO THE ACTUAL PLATE BEING USED.
USERS SHOULD ALSO TAKE CARE THAT NO POTENTIAL FLUORESCENT
OR LUMINESCENT CONTAMINATION LIES ON TOP OF THE PLATE. BE
AWARE THAT SOME PLATE SEALERS LEAVE BEHIND A STICKY RESIDUE
THAT MUST BE COMPLETELY REMOVED BEFORE STARTING
MEASUREMENTS.
CAUTION
BEFORE STARTING MEASUREMENTS, MAKE SURE THAT THE MICROPLATE
POSITION A1 IS INSERTED CORRECTLY. THE POSITION OF WELL A1 HAS
TO BE ON THE UPPER LEFT SIDE.
CAUTION
TO INSURE THE OPTIMAL WORKING OF TECAN INSTRUMENTS
WE RECOMMEND A SERVICE INTERVAL OF 6 MONTHS.
STOP
STOP
STOP

1. Safety
10 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 2016-04
It is assumed that the instrument operators, because of their vocational
experience, are familiar with the necessary safety precautions for handling
chemicals and biohazardous substances.
Adhere to the following laws 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 200
PRO MAY COME IN CONTACT WITH BIOHAZARDOUS/INFECTIOUS
MATERIAL. MAKE SURE THAT ONLY QUALIFIED PERSONNEL
OPERATE THE INSTRUMENT. IN CASE OF SERVICE OR WHEN
RELOCATING OR DISPOSING OF THE INSTRUMENT, ALWAYS
DISINFECT THE INSTRUMENT ACCORDING TO THE
INSTRUCTIONS GIVEN IN THIS MANUAL.

2. General Description
2016-04 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 11
2. General Description
2.1 Instrument
The Tecan Infinite 200 PRO is a multifunctional microplate reader with injector
option. The Infinite 200 PRO provides high performance for the vast majority of
today’s microplate applications and research and is robotic compatible.
2.1.1 Intended Use
The Infinite 200 PRO has been designed as a general purpose laboratory
instrument for professional use, supporting common 6 to 384-well microplates
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 on a selected set of assays only. It is the
responsibility of any operating authority to ensure that the Infinite 200
PRO has been validated for every specific assay used on the
instrument.
WARNING
LIMITATION:
WHEN USING THE GAS MODULE OPTION TO MEASURE AND
CONTROL OXYGEN LEVELS, THE INSTRUMENT IS INTENDED FOR
RESEARCH USE ONLY - NOT FOR USE IN DIAGNOSTIC
PROCEDURES.
WHEN OXYGEN LEVELS ARE NOT MEASURED OR CONTROLLED, THE
INSTRUMENT IS INTENDED FOR USE AS A GENERAL PURPOSE
LABORATORY INSTRUMENT FOR PROFESSIONAL USE.

2. General Description
12 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 2016-04
2.1.2 Multifunctionality
Depending on the mode of wavelength selection, the Infinite 200 PRO is
available in two different versions:
Infinite M200 PRO
Infinite F200 PRO
The following measurement techniques are supported by the Infinite M200 PRO:
Fluorescence Intensity (FI) Top
Fluorescence Intensity (FI) Bottom
Time-Resolved Fluorescence (TRF)
Fluorescence Resonance Energy Transfer (FRET)
Flash Fluorescence (with injectors)
Absorbance
Absorbance (with injectors)
Absorbance in cuvettes
Glow Luminescence
Flash Luminescence
Bioluminescence Resonance Energy Transfer (BRET)
A fully-equipped Infinite F200 PRO supports the following measurement
techniques:
Fluorescence Intensity (FI) Top
Fluorescence Intensity (FI) Bottom
Time-Resolved Fluorescence (TRF)
Fluorescence Resonance Energy Transfer (FRET)
Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET)
Flash Fluorescence (with injectors)
Fluorescence Polarization (FP)
Absorbance
Absorbance (with injectors)
Glow Luminescence
Flash Luminescence
Bioluminescence Resonance Energy Transfer (BRET)
AlphaScreen/AlphaLISA
Any common microplate ranging from 6 to 384 well formats conforming to the
ANSI/SBS standards (ANSI/SBS 1-2004; ANSI/SBS 2-2004, ANSI/SBS 3-2004
and ANSI/SBS 4-2004) may be measured with any of the above measurement
techniques. Switching between measurement techniques or plate formats is fully
automated via software. It is not necessary to manually reconfigure the optics in
order to switch between the reading modes supported by the Infinite 200 PRO.
Both instrument versions, the filter-based Infinite F200 PRO and the
monochromator-based Infinite M200 PRO, may be equipped with up to two
injectors.

2. General Description
2016-04 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 13
2.1.3 Performance
The Infinite 200 PRO has been designed to meet the requirements of a general-
purpose laboratory instrument.
The Infinite 200 PRO provides a range of parameters for optimizing the
measurement results according to: the assay type (cell-based or homogeneous),
the microplate type, and the dispensed volumes per well and dispensing speeds.
2.1.4 User Friendliness
The Infinite M200 PRO offers unparalleled flexibility in wavelength selection for
fluorescence intensity and absorbance measurements. Via software any
wavelength can be easily adjusted within the specified wavelength range. In
addition to single wavelength measurements, absorbance and fluorescence
spectra 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; slides containing fluorescence and absorbance
interference filters are easily accessible to the user.
Note
If the instructions given in this document are not correctly performed,
the instrument will either be damaged or the procedures will not be
performed correctly and the safety of the instrument is not guaranteed.
2.1.5 Onboard Control Button
The Infinite 200 PRO possesses an onboard control button to control plate
movements without the need to be connected to the software. Upon pressing the
‘Plate In/Out’ button, the current position of the plate carrier is automatically
recognized and the plate is moved into or out of the instrument.
Figure 1: Onboard of the Infinite 200 PRO. The ‘Plate In/Out’ button is located in the front
right corner of the top cover.

2. General Description
14 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 2016-04
2.1.6 Rear View
Figure 2: Rear panel
1 Instrument Fan
2 Main Power Switch
3 Main Power Socket
4 Label – RoHS Orange Logo
5 Label – Technical Inspection Agency (TÜV)
6 Power Supply Fan
7 Name Plate
8 Label – Options/Configuration
9 Injector Connection
10 USB Connection
11 Warranty Label: “ATTENTION
REMOVING OR BREAKING
THIS SEAL VOIDS
WARRANTY!”
CAUTION
ONLY TECAN AUTHORIZED SERVICE TECHNICIANS ARE ALLOWED
TO OPEN THE INSTRUMENT. REMOVING OR BREAKING THE
WARRANTY SEAL VOIDS THE WARRANTY.
STOP

2. General Description
2016-04 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 15
2.2 Software
The Infinite 200 PRO is delivered with the i-control software, for operating the
instrument and includes an online-help file and a printed Instructions for Use. The
software is formatted as a self-extracting archive on CD-ROM. (For information
about the system requirements, refer to the Instructions for Use for the i-control
software. The Instructions for Use for the i-control can be found on the software
CD.)
For advanced data reduction, the Magellan software can be used to control the
Infinite 200 PRO. Magellan offers all functionality for compliance with the FDA
regulation 21 CFR part 11 for electronic records and signatures (for more
information, contact your local 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
separate box, which feed one or two injector needles.
The injector needles are designed to inject liquid in any SBS-conform microplate
well types, in which the well-size is equal to or larger than an SBS standard 384-
well plate.
Figure 3: Injector-box with bottle holders
2.3.1 Injector Measurement Modes
The injectors of the Infinite 200 PRO can be used with the following measurement
modes:
•Fluorescence Intensity top and bottom
•Time Resolved Fluorescence
•Absorbance
•Flash Luminescence
•Glow Luminescence
•Dual Color Luminescence
As the measurement position is not the same as the injector position, a short time
delay (approx. < 0.5 s) between injection and reading occurs.
For details on how to set up a measurement with injectors, please refer to chapter
5.10.5 Injector.

2. General Description
16 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 2016-04
2.3.2 Injector Module Diagram
Figure 4: Schematic view of the injector module
2.3.3 Injector Pump Options
There are up to two pumps available for the Infinite 200 PRO (see Figure 4
above):
•Pump A feeds injector needle A
•Pump B feeds injector needle B
The 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 microplate well types, in which
the well-size is equal to or larger than an SBS standard 384-well plate.
•Two Injector Option (two pumps): Some applications, such as flash
luminescence reactions or dual reporter gene assays require the injection of
two independent liquids into the same well; therefore, Tecan Austria offers a
two-injector option.

2. General Description
2016-04 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 17
2.3.4 Storage Bottles and Bottle Holders
The injector box can accommodate up to two 125 ml bottles.
The standard bottle set supplied with the Injector option consists of:
•One 125 ml bottle and one 15 ml bottle for the “One Injector option”
(one pump) or
•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 bottles and tubes containing the fluids that are
to be injected can be attached securely to the holder using flexible PVC clasps.
The tubes from the injector syringe can be inserted into a carbon needle reaching
down to the bottom of the flask to ensure the optimal aspiration of even small
volumes of fluid.
Figure 5: Bottle holders

2. General Description
18 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 2016-04
2.3.5 Injector Carrier
The injector carrier, which includes the injector needles, can be easily removed
from the instrument for priming or washing the system and for optimizing the
injection speed.
Figure 6: Injector carrier
When using the injector during a measurement or for just dispensing a plate the
injector carrier must be inserted correctly into the instrument. Remove the injector
dummy and insert the carrier into the injector port. Press the carrier softly 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 correct.
If the injector carrier is not inserted correctly, the injector sensor does not
recognize the inserted carrier and neither dispensing nor injection is possible.
On the other hand, actions like washing and priming are enabled although the
injector carrier is inserted; therefore, always make sure that the injector carrier is
in the service position for washing and priming.
CAUTION
THE INJECTOR CARRIER MUST BE IN THE SERVICE POSITION
FOR WASHING UND PRIMING.
PRIME AND WASH MUST NOT BE PERFORMED
WHEN THE INJECTOR IS IN THE INSTRUMENT!
STOP

2. General Description
2016-04 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 19
Figure 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 THEREFORE WASHING AND PRIMING
WILL BE ENABLED, WHICH CAN DAMAGE THE INSTRUMENT.
STOP

2. General Description
20 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 2016-04
2.4 Measurement Techniques
The following sections provide an introduction to the Infinite 200 PRO
measurement techniques when fully equipped. To keep this compact, a few
simplifications have been made. For details see the references.
2.4.1 Fluorescence
The Infinite 200 PRO offers the basic fluorescence measurement technique and
some even more sophisticated variants:
•Fluorescence Intensity (FI) (or simply Fluorescence)
•Fluorescence Resonance Energy Transfer (FRET)
•Fluorescence Time Resolved (TRF)
•Fluorescence Polarization (FP)
FI may also be used to measure Fluorescence Resonance Energy Transfer
(FRET). For some microplate applications, FRET offers advantages over FI and
TRF, because they simplify assay preparation. These preferably apply for mix
and measure binding studies. Compared to FP, FRET requires both binding
partners to be labeled in a suitable way. On the other hand, FRET may utilize
TRF labels for increased sensitivity, then being referenced as HTRF
(Homogeneous TRF).
TRF should not be confused with Fluorescence Lifetime measurements.
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 (quantum of light). This is a part of the
energy, which has been absorbed before (electronic excitation), but could not be
released quickly enough into thermal energy.
The average time it takes between excitation and emission is called the
fluorescence lifetime. For many fluorescent molecular species, fluorescence
lifetime is on the order of nanoseconds (prompt fluorescence). After excitation,
fluorescence emission occurs with a certain probability (quantum yield), which
depends on the fluorescent species and its environmental conditions.
For a detailed treatise on fluorescence techniques and applications see:
Principles of Fluorescence Spectroscopy by Joseph R. Lakowicz, Plenum
Press.
A) Fluorescence Intensity (FI)
In many microplate applications, the intensity of fluorescence emission is
measured to determine the abundance of fluorescent labeled compounds. In
these assays, other factors having an influence on fluorescence emission need to
be controlled experimentally. Temperature, pH-value, dissolved oxygen, kind of
solvent etc. may significantly affect the fluorescence quantum yield and therefore
the measurement results.