Luminoskan Ascent
Luminoskan Ascent
User Manual
User Manual Rev. 2.1; Dec. 2003, Cat. no. 1507520
2 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
Table of Contents
Symbols and Markings ................................................................ 7
1. Introduction ....................................................................... 9
2. Instrument Layout............................................................ 11
3. Installation ....................................................................... 19
4. Operation ......................................................................... 31
5. Maintenance..................................................................... 55
6. Troubleshooting Guide..................................................... 65
7. Frequently asked questions (FAQ) about the Luminoskan
Ascent.............................................................................. 69
8. Instrument Service...........................................................71
9. Disposal of the Instrument and Materials........................ 77
10. Ordering Information ....................................................... 79
11. Glossary and Abbreviations .............................................83
12. Literature.......................................................................... 85
13. Warranty Certificate......................................................... 87
14. Specifications................................................................... 89
15. Index................................................................................. 95
Appendix A. Brief User’s Guide ................................................. 99
Appendix B. Addresses............................................................ 101
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 3
Contents
Symbols and Markings................................................................ 7
1. Introduction .......................................................................... 9
1.1 Intended use .................................................................................. 10
1.2 Method descriptions ..................................................................... 10
1.2.1 Luminometric measurement principle.................................. 10
2. Instrument Layout............................................................... 13
2.1 Front view ...................................................................................... 13
2.2 Rear view ....................................................................................... 14
2.3 Internal view .................................................................................. 15
2.4 Optical system............................................................................... 16
2.5 Control switches............................................................................ 16
2.6 Incubator........................................................................................ 17
2.7 Dispensers ..................................................................................... 17
2.8 Plate carrier.................................................................................... 18
3. Installation .......................................................................... 19
3.1 Upon delivery ................................................................................ 19
3.1.1 Unpacking............................................................................... 19
3.1.2 Checking delivery for completeness..................................... 20
3.1.3 Checking for damage during transport................................. 20
3.1.4 Environmental requirements and noise................................ 21
3.1.5 Things to avoid....................................................................... 21
3.1.6 Technical prerequisites.......................................................... 22
3.2 Releasing the transportation lock................................................. 22
3.3 Power and computer connections ............................................... 23
3.4 Installation of dispensers .............................................................. 25
3.5 Installation of the drop plate......................................................... 26
3.6 Plate adapters ................................................................................ 27
3.7 Adjusting the plate carrier............................................................. 29
3.8 Operational check.......................................................................... 30
4. Operation ............................................................................ 31
4.1 Switching on.................................................................................. 31
4.2 Loading the microplate ................................................................. 32
4.3 Luminometric measurement ........................................................ 33
4.3.1 Luminometric scaling............................................................. 34
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User Manual Rev. 2.1, Cat. no. 1507520
4.4 Other functions...............................................................................35
4.4.1 Orbital shaking ........................................................................35
4.4.2 Incubator .................................................................................38
4.4.3 Dispensers...............................................................................38
4.4.4 Dispenser head height adjustment ........................................41
4.4.5 Chemical resistance of the dispenser ...................................43
4.5 Changing the measurement direction ..........................................46
4.6 Installing or removing the light shield ..........................................49
4.7 Installing the filters .........................................................................50
4.8 Shutdown .......................................................................................53
5. Maintenance........................................................................ 55
5.1 Routine cleaning of the instrument...............................................55
5.2 Cleaning the optical system ..........................................................56
5.2.1 Visual filter check ....................................................................56
5.3 Cleaning the plate carrier...............................................................57
5.4 Replacing the fuses........................................................................58
5.5 Routine maintenance of optional dispensers and main lens ......59
5.5.1 Basic maintenance..................................................................59
5.5.2 Extended maintenance...........................................................59
5.5.2.1 Weak detergent or 10% bleach ....................................................59
5.5.2.2 Weak acid and base in sequence..................................................60
5.5.2.3 Cleaning the main lens..................................................................60
5.6 Periodic maintenance ....................................................................61
5.6.1 Replacing the dispenser tubings............................................61
5.6.2 Replacing the dispensing tip ..................................................63
5.6.3 Replacing the dispenser syringe............................................64
6. Troubleshooting Guide ........................................................ 65
6.1 Troubleshooting.............................................................................65
6.2 Error messages ..............................................................................68
7. Frequently asked questions (FAQ) about the Luminoskan
Ascent ................................................................................. 69
8. Instrument Service .............................................................. 71
8.1 Service request protocol ...............................................................71
8.2 Decontamination procedure..........................................................71
8.3 Certificate of decontamination ......................................................73
8.4 Shipping the instrument (or items) ...............................................75
8.5 Service contracts............................................................................75
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 5
9. Disposal of the Instrument and Materials........................... 77
9.1 Disposal of the instrument............................................................ 77
9.2 Disposal of materials..................................................................... 77
10. Ordering Information ....................................................... 79
10.1 Product code numbers ................................................................. 79
10.2 List of recommended spare parts ................................................ 82
10.3 Ordering filters............................................................................... 82
11. Glossary and Abbreviations ............................................. 83
12. Literature ......................................................................... 85
13. Warranty Certificate ........................................................ 87
13.1 Warranty limitations ...................................................................... 88
14. Specifications .................................................................. 89
14.1 General specifications................................................................... 89
14.2 Safety specifications ..................................................................... 91
14.3 In conformity with the requirements............................................ 92
14.4 Performance specifications........................................................... 93
15. Index ................................................................................ 95
Appendix A. Brief User’s Guide ................................................ 99
Appendix B. Addresses ........................................................... 101
6 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
Symbols and Markings
SYMBOLS USED IN THE LUMINOSKAN ASCENT
Power ON
Power OFF
Connection to the protective grounding system
WARNING MARKINGS USED ON THE INSTRUMENT
Caution: risk of electric shock.
Caution: risk of personal injury to the operator or a
safety hazard to the surrounding area. See the
WARNING MARKINGS USED IN THE DOCUMENTATION
accompanying documentation.
CE compliance mark
Caution: risk of electric shock.
Caution: risk of personal injury to the operator or a
safety hazard to the surrounding area.
Caution: risk of serious damage to the instrument, other
equipment or loss of performance or function in a
specific application.
Caution: biohazard risk.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 7
Luminoskan Ascent, Cat. no. 5300160 and 5300170 (with dispenser)
User Manual Rev. 2.1; Dec. 2003, Cat. no. 1507520
Copyright
1995 – 2003 Thermo Electron Corporation. All rights reserved. Printed in
Finland. Reproduction of the accompanying user documentation in whole
or in part is prohibited.
Patents
Luminoskan Ascent has national and international patents and patents
pending.
Trademarks (
and )
Ascent Software, Luminoskan, Luminoskan Ascent and Microtiter are
registered trademarks of Thermo Electron. BioBind and Microlite are
trademarks of Thermo Electron. Virkon is a registered trademark of Antec
International Ltd. All other trademarks, registered trademarks and
copyrights are property of their respective holders.
Disclaimer
Thermo Electron 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. Although this manual has been
prepared with every precaution to ensure accuracy, Thermo Electron
assumes no liability for any errors or omissions, nor for any damages
resulting from the application or use of this information. This manual
supersedes all previous editions.
No liability for consequential damages
Thermo Electron shall not be liable for any damages whatsoever arising
out of the use or inability to use this product.
Contact information
P.O. Box 100, FIN-01621 Vantaa, Finland
Tel. +358-9-329 100, Fax +358-9-3291 0415
www.thermo.com
8 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
1. Introduction
The Luminoskan Ascent (Fig. 1) designed and made by Thermo Electron is
a microplate luminometer which offers versatility and flexibility for even
the most demanding luminometric applications. The instrument covers the
full range of glow and flash luminometric applications.
Fig. 1. Luminoskan Ascent
Ascent Software controls all the reader functions and provides easy assay
optimization, flexible data handling and convenient report formatting. The
software for the Luminoskan Ascent is a dedicated software for
luminometric applications.
High sensitivity is one of the main benefits of the instrument. The
Luminoskan Ascent can also be equipped with filters for luminometric
applications.
Up to three reagent dispensers can be fitted on-board, making the reagent
addition simple and highly accurate. The ability of the instrument to
dispense and measure simultaneously enables the detection of flash
luminescence reactions and rapid kinetic applications. For assays requiring
temperature control, the instrument has an on-board incubator. Built-in
orbital shaking speeds up reaction times and ensures effective mixing.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 9
The robotic integration is simple and effective with the Luminoskan
Ascent. The plate carrier allows convenient access for the robotic arm and
Ascent Software is easy to integrate with robotic and HIS/LIMS systems.
The Luminoskan Ascent is also fully compatible with Thermo Electron
robotic plate handling devices, expanding the measurement capacity. For
further information, contact your local Thermo representative.
1.1 Intended use
1. The Luminoskan Ascent is a high-quality microplate luminometer
intended for laboratory research use by professional personnel. The
Luminoskan Ascent is used to measure luminescence from suitable 1to 384-well plates mentioned in this manual. It also has incubation,
shaking and reagent dispensing capabilities.
2. For verification of the entire system, it is recommended that Good
Laboratory Practices (GLP) be followed to guarantee reliable analyses.
3. Use for self-testing is excluded.
1.2 Method descriptions
1.2.1 Luminometric measurement principle
The filter slots 7 and 8 are reserved only for luminometric measurements.
Filter slot 7 is empty for measurement and filter slot 8 is blocked to enable
measurement of the PMT (photomultiplier tube) dark current. This feature
is important to obtain optimal sensitivity. The light path from the
measurement well to the first lens is protected by a light shield (Fig. 4.6a
and Fig. 4.6b). The light shield is required with 96- and 384-well plates to
avoid crosstalk in luminometric measurements. When plates are higher
than 15 mm, the light shield must be removed. If the 384-well plate has a
height of less than 15 mm, then an adapter must be used below the plate
to raise the plate to the proper height. This adapter for low 384-well plates
(Fig. 3.6:4) is included in the Luminoskan Ascent instrument and can also
be ordered separately.
10 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
Caution: A luminometric measurement without a light
shield may cause extra crosstalk.
Caution: The light shield must be removed with higher
plates.
Note: The emission filter slots 7 and 8 are reserved only for
luminometric measurements. Filter slot 7 is empty for
measurement and filter slot 8 is blocked to enable
measurement of the PMT (photomultiplier tube) dark
current.
DO NOT install any filters into slots 7 and 8.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 11
12 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
2. Instrument Layout
2.1 Front view
5
1
2
3
4
1 Instrument housing
2 Dispenser and optics cover
3 Instrument chassis
4 Measurement chamber door
5 Power, busy and error indicator
Fig. 2.1 Luminoskan Ascent front view
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 13
2.2 Rear view
6
7
5
4
3
1
2
1 Mains power supply socket
2 Power switch (ON/OFF)
3 Power fuses
4 Serial communication connector for the computer
5 Identification plate
6 Housing retaining screws
7 Cooling-air outlet
Fig. 2.2 Luminoskan Ascent rear view
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User Manual Rev. 2.1, Cat. no. 1507520
2.3 Internal view
1
7
2
3
5
6
4
8
1 Light cover for the optical unit
2 Dispensers (opt.)
3 Dispensing head (opt.)
4 Leakage tray (opt.)
5 Dispensing head holder (opt.)
6 Dummy plug
7 Control switches
8 Cover sensor
Fig. 2.3 Luminoskan Ascent internal view
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 15
2.4 Optical system
2
3
5
4 1
1 In the Luminoskan Ascent the light beam is strictly limited by a light shield to
avoid crosstalk.
2 Mirror
3 An optional filter in the filter wheel
4 The photomultiplier tube (PMT) detects the light.
5 Light shield
Fig. 2.4 Principle of the optical system
2.5 Control switches
The control switch box (Fig. 2.3:7) contains three rocker switches for
priming and emptying dispenser tubings, one rocker switch for driving the
plate carrier in or out, and a sensor (Fig. 2.3:8) to monitor that the
dispenser cover is in the closed position in luminometric measurements.
The priming control switches are only functional when the plate carrier is
located outside the instrument.
16 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
2.6 Incubator
The incubator contains two heating element plates in the measurement
chamber, one heating element plate under the microplate and another
above it. The incubator only heats, but does not cool.
2.7 Dispensers
The optional dispensers, 1 to 3 dispensers from left to right, are located
inside the instrument housing under the dispenser cover, as seen in
Fig. 2.3. The dispensers consist of modular digital pumps with valves,
syringes, tubing and dispensing heads. The dispensing heads have three
alternative dispensing positions, one of these dispenses into the well in the
measurement position.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 17
2.8 Plate carrier
p
A1 corner of the
micro
late
2
1 Positioning lever
2 Adjustable stoppers
3 Waste strip holder (opt., 4 wells) for the tip priming during the measurement
session
4 Plate carrier door
3
1
4
Fig. 2.8 Plate carrier
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User Manual Rev. 2.1, Cat. no. 1507520
3. Installation
3.1 Upon delivery
3.1.1 Unpacking
The Luminoskan Ascent
Move the unpacked instrument to its site of operation. To prevent
condensation, the instrument should be left in its protective plastic
wrapping until the ambient temperature has been reached. Unpack the
Luminoskan Ascent instrument and accessories carefully with the arrows
on the transport package pointing upwards. Open the top of the package
and lift the Luminoskan Ascent
(Fig. 3.1). The following notes and instructions are sent with the instrument
and are immediately available when you open the package:
• the packing instructions
• the packing list
• the Thermo Electron Warranty Certificate card
• the performance test reports
• the User Manual.
Caution: DO NOT touch or loosen any screws or parts other
than those specifically designated in the instructions. Doing so
might cause misalignment and will void the instrument
warranty.
Caution: The Luminoskan Ascent
(46 lbs.) without dispensers and should be lifted with care. It is
recommended that two persons lift the instrument together,
taking the proper precautions to avoid injury.
is packed in a specially designed shipping carton.
out of the shipping carton
weighs approximately 21 kg
Retain the original packing materials and shipping carton for future
transportation. Also retain all the documentation provided with the
instrument.
If you relocate your instrument or ship it for service, remember to:
1. Empty the dispenser(s) and remove the tube assembly.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 19
2. Remove any loose items from the plate carrier, for example, adapters,
plates and priming vessels.
3. Remove the power cable as well as the serial cable.
4. Replace the transportation lock.
For further information, see Section 8.4 Shipping the instrument (or items).
Intended lifting
area
Fig. 3.1 Luminoskan Ascent
3.1.2 Checking delivery for completeness
Check the enclosed packing list against order. If any parts are missing,
contact your local Thermo representative or Thermo Electron Oy.
3.1.3 Checking for damage during transport
Visually inspect the transport package, the instrument and the accessories
for any possible transport damage.
If the carton 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.
20 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
Visually check all interconnections in the basic instrument. Check that
there are no loose parts inside the instrument.
If any parts are damaged, contact your local Thermo representative or
Thermo Electron Oy.
3.1.4 Environmental requirements and noise
When you set up your Luminoskan Ascent, avoid sites of operation with
excess dust, vibrations, strong magnetic fields, direct sunlight, draft,
excessive moisture or large temperature fluctuations.
• Make sure the working area is flat, dry, clean and vibration-proof and
leave additional room for cables, connections, controlling computer,
printer, etc.
• Make sure the ambient air is clean and free of corrosive vapors, smoke
and dust.
• Make sure the ambient temperature range is between +10°C (50°F)
and +40°C (104°F).
• Make sure relative humidity is between 10% and 90% (noncondensing).
Leave sufficient space (at least 10 cm) at both sides of the instrument and
at the back of the unit to allow adequate air circulation. Make space for the
controlling computer on one side of the Luminoskan Ascent.
The Luminoskan Ascent does not produce operating noise at a level which
could be harmful. No sound level measurements are needed after
installation.
Warning: DO NOT operate the instrument in an environment
where potentially damaging liquids or gases are present.
3.1.5 Things to avoid
DO NOT smoke, eat or drink while using the Luminoskan Ascent. Wash
your hands thoroughly after handling test fluids. Observe normal
laboratory procedures for handling potentially dangerous samples. Use
proper protective clothing. Use disposable gloves. Be sure the working
area is well-ventilated.
Never spill fluids in or on the equipment.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 21
3.1.6 Technical prerequisites
Place the instrument on a normal laboratory bench close to the mains
power supply socket. The net weight of the unit is approx. 21 kg (46 lbs.).
The instrument operates at voltages of 100 – 240 Vac. The frequency range
is 50/60 Hz.
3.2 Releasing the transportation lock
2
3
4
1
1 Cover retaining screws, 4 pieces
2 Dispenser and optics cover
3 Rear of the cover
4 Point to where cover lifted
Fig. 3.2a Removing the instrument cover
1. Remove the four cover retaining screws (Fig. 3.2a:1).
2. Open the dispenser and optics cover (Fig. 3.2a:2).
3. Lift the rear of the cover at first about 3 cm (Fig. 3.2a:3).
4. Lift the cover aside (Fig. 3.2a:4).
5. Undo the two screws (Fig. 3.2b:1) at the right rear corner of the
measurement chamber.
6. Turn the locking piece upside down (Fig. 3.2b:2).
22 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
7. Fit the locking piece back with the fitting screws (Fig. 3.2b:3).
8. Refit the cover by first fixing the front corners.
1 2
3
1 Connected locking piece with two screws
2 Disconnected locking piece turned upside down
3 Reconnected locking piece
Fig. 3.2b Removing the transportation lock
Caution: If the locking piece is not fitted into its place, light
may enter the measurement chamber and affect the results.
3.3 Power and computer connections
1. Ensure that the power switch (Fig. 3.3:1) is in the OFF position.
2. Connect the mains supply cable to the mains power supply socket
(Fig. 3.3:2). The instrument box contains two different mains supply
cables with North American and European types of plugs. Select the
correct type used in your laboratory. If any other type of mains supply
cable is needed, use only cables certified by the local authorities.
3. Connect the instrument to a correctly installed line power outlet that
has a protective conductor also called ground or earth.
4. Connect the serial cable to the serial connector (Fig. 3.3:3) and secure it
with the locking screws. Connect the other end similarly to the
controlling computer.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 23
3
2
1
1 Power switch (ON/OFF)
2 Mains power supply socket
3 Serial connector
Fig. 3.3 Power and computer connections
Warning: Always make sure the power switch on the
instrument is in the OFF position and remove the mains
power supply cable from the back of the instrument prior
to any installation or relocation of the instrument.
Warning: Never operate your instrument from a power
outlet that has no ground connection.
24 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
3.4 Installation of dispensers
The optional dispensers 1 to 3 are installed in number order from left to
right. The complete dispensing assemblies are packed with the
accessories.
hole of the valve. Ensure that the aspirate tubing is finger-tight. If
necessary, turn the fitting another quarter to half turn using a 7.9 mm (5/16
in.) wrench. The aspirate tubing is used to fill the syringe with reagent.
When using the dispensers, make sure that the aspiration tube end is
completely submerged in the reservoir and there is a sufficient volume of
the reagent in the reservoir (for all primings and the actual dispensing).
Fit the complete dispensing tube assembly (Fig. 3.4:2) into the right hole of
the valve and tighten it finger-tight. Then turn the fitting another quarter to
half turn using a 7.9 mm (5/16 in.) wrench. The dispensing tube is used to
dispense reagent from the syringe into a microplate. Place the dispensing
heads in the dispensing head holder on the left-hand side of the
dispensers.
First push the plunger manually upwards into the upper position before
tightening the plunger lock screw (Fig. 3.4:5). Ensure that the plunger lock
screw is sufficiently tightened. Note that the plunger can be extremely
firm.
1
The aspirate tubing (Fig. 3.4:1) is factory installed into the left
1
Instructions concerning the pump are reproduced from CAVRO XP3000 Modular Digital
Pump Operators Manual made by Cavro Scientific Instruments, Inc., USA, 1994.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 25
1
2
3
4
5
1 Aspirate tube assembly (incl. tubing and end weight)
2 Complete dispensing tube assembly
3 3-port valve
4 Dispensing syringe (1.0 ml) and plunger
5 Plunger lock screw
Fig. 3.4 Automatic dispenser unit
3.5 Installation of the drop plate
The instrument is supplied with a special drop plate (Fig. 3.5a). The drop
plate is used to protect the instrument from damage caused by accidental
dispensing without any microplate. If the user forgets to place a microplate
onto the plate carrier but has the drop plate in place, the reagent will be
dispensed into the drop plate, not inside the instrument. The drop plate is
placed like an adapter into the plate carrier (Fig. 3.5b) and the microplate is
placed onto the drop plate. The holding capacity of the drop plate is 19 ml
of liquid.
Note: The drop plate cannot be used for bottom reading.
26 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
Fig. 3.5a Drop plate
Fig. 3.5b Drop plate placed into the plate carrier
3.6 Plate adapters
Reading certain plate types with the Luminoskan Ascent instrument
requires special plate adapters (Fig. 3.6). These plate adapters are required
for the following reasons:
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 27
1. Raising the 384-well plate with a height of less than 15 mm to the
proper height for luminometric measurement.
2. If the 384-well plate height is less than 13.5 mm, an adapter must be
used to raise the plate for dispensing.
3. Positioning of other sample vessel types than microplates (for
example, Petri dishes or Terasaki plates) onto the plate carrier for
measurement and dispensing.
4. Supporting flexible PCR plates and tubes on the plate carrier for
measurement and dispensing.
When using any adapter, measure the height of the plate and the adapter
together and ensure that this value is correctly entered into the Ascent
Software plate template parameter file. Refer to the Ascent Software
User’s Guide for further information.
1
2
1 Adapter for Petri dish, 6 x 40 mm
2 Adapter for 96-well PCR plates
3 Adapter for Petri dish, 93 mm
4 Adapter for low 384-well plates
Fig. 3.6 Example of plate adapters
28 Luminoskan Ascent
3
4
User Manual Rev. 2.1, Cat. no. 1507520
Note: If the orientation of the plate adapter is important, the correct
orientation is printed on the adapter or the adapter has been
designed in such a way that only one orientation is possible.
When the adapter is needed, place it into the plate carrier. Then place the
sample vessel(s) on the corresponding adapter or a drop plate, select the
correct plate template (marked “with adapter”) from Ascent Software and
run the instrument normally.
Note: Always remember to remove the plate adapter before
using the instrument with any other plate types.
3.7 Adjusting the plate carrier
The plate carrier of the Luminoskan Ascent has been designed for plates
with different footprints and to be robot compatible. The plate carrier has
two adjusting knobs with two different orientations. These knobs have
been set for standard plate types at the factory. The setting is also valid for
the robotic 384-adapter but readjustment of the knobs will be needed with
the plate types requiring special adapters (see Section 3.6). Fig. 3.7 shows
how the knobs should be adjusted for special plate adapters. Both knobs
should be rotated 90° clockwise after which all special adapters fit into the
plate carrier.
Rotate two knobs
90° clockwise
Fig. 3.7 Two plate carrier knobs rotated 90
°
clockwise for special plate
adapters
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 29
3.8 Operational check
Switch the instrument ON. The instrument automatically performs a
complete set of initialization tests or adjustments.
When the initialization tests and adjustments have been completed
successfully, the indicator LED turns from yellow to green.
If anything fails in the initialization tests or adjustments, the indicator LED
will turn red. In this case, switch the instrument OFF and ON again. If this
failure is repeated, contact authorized technical service.
The instrument is ready for operation.
The instrument also performs different kinds of additional runtime
hardware tests before each measurement run, like automatic blanking.
Because of the relative nature of luminometry, we recommend you use
known samples or controls to verify proper instrument operation.
30 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
4. Operation
The Luminoskan Ascent is fully computer-controlled. Thermo Electron
Ascent Software controls the reader functions and provides complete data
handling and report formatting. For further details on how to use the
software, refer to the Ascent Software User’s Guide.
Warning: DO NOT use any other software than Thermo
Electron software designed for this instrument. Use only valid
combinations of the PC software and the instrument’s internal
EEPROM.
Warning: The Luminoskan Ascent instrument does not verify
the logic flow of the received commands.
The instrument is equipped with a power switch (ON/OFF) and a threecolor LED indicator. When the instrument is switched ON, the color
indicates the state of the instrument:
Green The instrument is ready and waiting for a command.
Orange The instrument is busy, executing a command.
Red The instrument has found an error, the error message is sent
to the computer and the computer has not acknowledged it.
4.1 Switching on
When switched on, the instrument performs the initialization tests. When
all the initialization tests and adjustments have been performed
successfully, the indicator LED turns green and the instrument is ready to
receive computer commands. The recommended warm-up time is
15 minutes, but the instrument will perform commands immediately after
the initialization period.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 31
4.2 Loading the microplate
The microplate is loaded onto the plate carrier of the instrument for
measurement. The plate carrier is able to handle microplates of different
sizes, therefore the free space in the plate carrier is clearly larger than the
standard 96-well plate. The positioning lever (Fig. 4.2:1) in the plate carrier
(Fig. 4.2) will automatically position the plate correctly into the upper left
corner of the carrier when the plate is driven in.
A1 corner of the
microplate
Positioning lever
Fig. 4.2 Plate carrier
32 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
4.3 Luminometric measurement
Luminometric reading is optimized for 96- and 384-well plates and top
reading. To obtain the best sensitivity and the lowest crosstalk, a light
shield (Fig. 4.6a) between the optics and the plate must be used. In
luminometric measurements the dispenser cover must be closed.
Reading 1- to 48-well plates as well as bottom reading can cause increase
in background signal and effect sensitivity in luminometric measurements,
because the light shield cannot be used and most of these plates are
transparent, which enables light leakage from one well to another.
A measurement function has several phases:
1. The anode voltage of the PMT is set according to the selected value of
the software.
2. The filter slot 8 (= block) is held in place until the plate door is closed.
The empty position/selected filter is then driven to the measurement
positions. The filter slot 8 is also used for the blanking procedure to
compensate for the possible PMT drift.
3. In the blanking procedure the instrument reads the PMT dark signal. If
the bottom value drifts, the results are compensated. To obtain the
most accurate results, the measurement time of the dark must be as
long as the measurement time of the sample. The measurement time of
the dark is divided into two parts. The first half is measured before the
sample measurement and the second half after the sample
measurement. The time used for the instrument blanking procedure
can be selected from Ascent Software (refer to the Overview part of the
Ascent Software User’s Guide).
Note: Ensure that there are no items preventing the closure of
the plate carrier door when measurement is started. DO NOT
open the plate carrier door during measurement.
Note: DO NOT install any filters into the filter wheel positions
7 and 8.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 33
4.3.1 Luminometric scaling
The measured results are expressed as Relative Light Units (RLU). Scaling
is a way to convert readings to show desired values.
1. Prepare a scaling reference solution and a blank solution and pipette
several wells of blanks and the known concentration.
2. Measure the wells using the correct filter or an open slot.
3. Calculate the average values of the measured reference values and
blank values. The blank values should be very small compared to the
reference values.
4. Calculate the scaling factor using calculated average values:
Factor = Known reference / (Measured reference - Measured blank)
Example
• The known concentration is 500 pmol/well.
• The calculated average of the measured reference wells is
1825 RLU.
• The calculated average of the measured blank wells is 0.2 RLU.
• Factor = 500 / (1825 - 0.2) = 0.274
• After scaling the measured result is: 0.274 x 1825 RLU =
500 pmol/well.
You can find the scaling entry in Ascent Software. The menu selection is
Setup|Filters. Select the corresponding filter or None and key in the
scaling factor. The measured results are expressed as Relative Light Units
(RLU).
34 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
4.4 Other functions
4.4.1 Orbital shaking
The track movement system can perform the shaking action. The speed is
adjustable from 60 to 1200 rpm (revolutions per minute) and the diameter
of the orbital movement is adjustable from 1 to 50 mm.
Some combinations of speed and diameter would cause too high g-forces
inside the well area resulting in spills inside the measurement chamber.
Therefore, only certain combinations are available.
The following tables (Table 4.4.1a – Table 4.4.1e) show the recommended
and the not recommended but available and unavailable speed and
diameter combinations with different plate types. These tables are based
on the liquid used being of low viscosity like water and the volumes being
appropriate (the wells are not full). The gray area in the tables show the
recommended (Recommended), the black area the not recommended
(Possible), and the white area the prevented speed and diameter
combinations (Not allowed).
Table 4.4.1a
1200
1140
1080
1020
960
900
840
780
720
660
600
540
480
Speed (rpm)
420
360
300
240
180
120
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 35
Shaking speeds for 48- (or more) well plates
Rec omm ended Not a llowed
60
0
1 3 5 7 9 111315171921232527 2931333537394143454749
Diameter (mm)
Table 4.4.1b
1200
1140
1080
1020
960
900
840
780
720
660
600
540
480
Speed (rpm)
420
360
300
240
180
120
60
0
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49
Shaking speeds for 24-well plates
Recommended Possible Not allowed
Diameter (mm)
Table 4.4.1c
1200
1140
1080
1020
960
900
840
780
720
660
600
540
480
Speed (rpm)
420
360
300
240
180
120
60
0
135791113151719212325272931333537394143454749
Shaking speeds for 12-well plates
Diameter (mm)
36 Luminoskan Ascent
Rec ommended Possible Not allowed
User Manual Rev. 2.1, Cat. no. 1507520
Table 4.4.1d
1200
1140
1080
1020
960
900
840
780
720
660
600
540
480
Speed (rpm)
420
360
300
240
180
120
60
0
1 3 5 7 9 111315171921232527293133353739 4143454749
Shaking speeds for 6-well plates
Recommended Possible Not allowed
Diameter (mm)
Table 4.4.1e
1200
1140
1080
1020
960
900
840
780
720
660
600
540
480
Speed (rpm)
420
360
300
240
180
120
60
0
1 3 5 7 9 11 131517192123 252729 3133353739414345 4749
Shaking speeds for 95 mm Petri dish
Recommended Possible Not allowed
Diameter (mm)
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 37
4.4.2 Incubator
The incubator contains two heating element plates in the measurement
chamber, one heating element plate under the microplate and another
above it. Both heating element plates are temperature-controlled. The
upper plate is slightly warmer than the lower plate to avoid condensation
on the plate lid. The measurement chamber is large to adopt different plate
formats and therefore extensive evaporation may cause some variations in
the temperatures between the wells. Consequently, when using
incubations with extended periods of time in the instrument, a plate lid is
recommended.
If the incubation period is long without any other important functions, the
Luminoskan Ascent automatically changes the place of the plate within the
measurement chamber to minimize temperature differences between the
wells.
4.4.3 Dispensers
The optional dispensers, 1 to 3 dispensers numbered from left to right, are
located inside the instrument housing under the dispenser cover
(Fig. 2.1:2). The dispensing heads have three alternative dispensing
positions, one of these dispenses into the well in the measurement
position (M). The positions are optimized for a 96-well plate (Fig. 4.4.3a).
When the dispensers are not in use, the dispensing heads may be stored
in the dispensing head holder, but the tip holes to the measurement
chamber must be closed with dummy plugs.
Note: Ensure that the tip position in the instrument
corresponds to that defined in Ascent Software.
38 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
1
2 2
1
Y
M
X
1 M position: Dispensing head directed in the measurement well position.
Dispensing into the well during the measurement step is possible with this
position.
2 Y position: Dispensing head directed in the well next to the measurement
position in the Y direction. When this position is used for dispensing, an extra
plate movement is carried out before the measurement step causing minor
time delays.
3 X position: Dispensing head directed in the well next to the measurement
position in the X direction. When this position is used for dispensing, an extra
plate movement is carried out before the measurement step causing minor
time delays.
3
3
Fig. 4.4.3a Dispenser tip positioning optimized for a 96-well plate
Before inserting the dispensing head into a dispensing position, prime the
syringe and tubing to an external priming vessel. The instrument has no
internal priming vessel for priming the syringe and tubing. You can find
the priming instructions in the Ascent Software User’s Guide. The
minimum priming volume needed is 700 µl and the recommended volume
is 2700 µl.
The Luminoskan Ascent also has control switches for priming the
dispenser tubing. With these switches, priming can be carried out
alternatively by using an external priming vessel or by using an empty
microplate on the plate carrier as a priming vessel or a drop plate. When
priming is performed with the dispensing heads installed into the M, X or
Y positions, place the empty priming vessel into the plate carrier. Initiate
priming by pressing the corresponding switch when the plate carrier is
located outside the unit. Priming is carried out as long as the
corresponding switch is pressed.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 39
Note: The priming control switches are functional only when
the plate carrier is located outside the instrument. DO NOT use
priming vessels higher than the actual plate intended to be
used in the assay. Notice the dispensing height adjustment.
Priming with Ascent Software must always be performed with
the dispensing heads removed from the M, X or Y dispensing
positions.
Note: Never use liquids that can cause any precipitation,
clotting or contain any mechanical particles with the automatic
dispensers.
The instrument has a Prime Tip feature. If this function is selected in the
Dispense or Dispense And Measure steps in Ascent Software, the
dispenser dispenses about 5 µl reagent into the tip priming vessels every
time the instrument fills the syringe. This makes the volume of the first well
equal to that of the others. We recommend you use the tip priming feature
when the dispensing volumes are small, for example, 5 – 20 µl. Notice that
the Execute by command causes the syringe to fill.
A1 corner of the
microplate
Fig. 4.4.3b Tip priming vessel (1) is a piece of a breakable strip
40 Luminoskan Ascent
1
User Manual Rev. 2.1, Cat. no. 1507520
The recommended tip priming vessel consists of four wells of a breakable
96-well plate strip (Fig. 4.4.3b). There is a holder for the tip priming vessel
in the right rear corner of the plate carrier. The four-well piece of a strip
should be exchanged after about 300 tip primings.
You may need to adjust the dispenser speed. The default setting is for a
water-based liquid. You can find the adjustments and selections in Ascent
Software.
When dispensing is started, the liquid volume in the well should be less
than half of the total well volume (for example, the volume should be less
than 200 µl in a typical 96-well plate).
4.4.4 Dispenser head height adjustment
Plate height 13.5 – 15.0 mm,
e.g., 96-well plate
Plate height 15.1 – 18.0 mm,
e.g., 48-well plate
Plate height 18.1 – 21.0 mm,
e.g., 24- to 6-well plates
No adjustment collars
One adjustment collar
Two adjustment collars
Fig. 4.4.4a Dispenser head height
The correct dispenser head height (Fig. 4.4.4a) is very important to avoid
contaminating neighboring wells. The correct tip height is also important
to prevent damaging the tip or the plate.
Note: Ensure that the dispenser tips are always correctly
inserted sufficiently deep into their slots.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 41
The plate height is one of the template parameters in Ascent Software and
it is defined as the height of the uncovered well from the bottom of the
plate, not the inside height of the well. The selected dispenser head height,
the used plate and the template selected in Ascent Software must match
to avoid problems. To see the heights of plates in Ascent Software, select
Setup and edit Plate Templates.
Some 384-well plates are lower than the standard 96-well plates. If the
plate height is less than 13.5 mm, an adapter must be used to raise the
plate. Measure the height of the plate and the adapter together and enter
this value to the plate template.
Caution: Plate manufacturers may change the dimensions of
plates without any prior notice or change in order numbers.
Check the dimensions when you start using plates from a new
box.
The dispenser head height is adjusted with red adjustment collars by
moving them from either side of the fixed stopper collar (Fig. 4.4.4b).
1
2
1 Dispenser head tube
2 Red adjustment collars
Fig. 4.4.4b Changing the position of the red adjustment collars
1. Remove the dispenser head tube (Fig. 4.4.4b:1) from the brass tube
lock.
2. The red adjustment collars (Fig. 4.4.4b:2) should be moved from one
side of the fixed collar to the other to select the correct dispenser head
height.
3. Fit the dispenser head tube back into place.
42 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
4.4.5 Chemical resistance of the dispenser 1
The following table (Table 4.4.5) is intended to provide guidelines for
compatibility with materials used in the fluid path of the dispensers.
Compatibility information is based on charts provided by the material
manufacturer. Cavro recommends that each laboratory determines
compatibility for their respective applications.
Caution: Failure to determine compatibility of chemicals used
in individual applications with the XP 3000, may result in
damage to the dispenser and/or test results.
Plastic materials used in dispensers:
Polysulfone: Cross Flow Manifold Assembly in the aspirate syringe
Teflon (PTFE, TFE, FEP): tubing; valve plug, and seal
Kel F: valve body
Polypropylene: fittings for tubing, and dispensing tip
Note: Also take into account the chemical resistance of
microplates.
Classification in the table:
– No data available
0 No effect – excellent
1 Minor effect – good
2 Moderate effect – fair
3 Severe effect – not recommended
1
Instructions concerning the pump are reproduced from CAVRO XP 3000 Modular Digital
Pump Operators Manual made by Cavro Scientific Instruments, Inc., USA.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 43
Table 4.4.5 Compatibility chart of materials suitable with the dispenser
Solvent Polysulfone Teflon Kel F Polypropylene
Acetaldehyde – 0 0 0
Acetates – – 0 0
Acetic Acid 0 0 0 0
Acetic Anhydride – – 0 –
Acetone 3 0 0 0
Acetyl Bromide – 0 – –
Ammonia 0 0 – 0
Ammonium Acetate – 0 – –
Ammonium Hydroxide – 0 0 0
Ammonium Phosphate – – 0 0
Ammonium Sulfate – – 0 0
Amyl Acetate – 0 – 3
Aniline – 0 0 0
Benzene 3 0 3 *
Benzyl Alcohol – 0 0 0
Boric Acid – 0 0 0
Bromide – 0 0 *
Butyl Alcohol 2 0 0 1
Butyl Acetate 3 0 – *
Carbon Sulfide – 0 – *
Carbon Tetrachloride 0 0 1 3
Chloroacetic Acid – 0 0 –
Chlorine – 0 1 3
Chlorobenzene 3 – – 3
Chloroform 3 0 – 3
Chromic Acid 3 0 0 –
Cresol – 0 – *
Cyclohexane 0 0 – 3
Dimethyl Sulfoxide (DMSO) 0 0 0 0
Ethers – 0 – **
Ethyl Acetate 3 0 – 0
Ethyl Alcohol 0 0 – 0
Ethyl Chromide – 0 1 3
Continued
44 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
Cont.
Solvent Polysulfone Teflon Kel F Polypropylene
Formaldehyde 0 0 0 0
Formic Acid – 0 0 0
Freon 2 0 2 0
Gasoline 2 0 0 3
Glycerine 0 0 0 0
Hydrochloric Acid 0 0 0 0
Hydrochloric Acid (conc.) 0 0 0 0
Hydrofluoric Acid 2 0 0 *
Hydrogen Peroxide – 0 0 0
Hydrogen Peroxide (conc.) – 0 0 0
Hydrogen Sulfide – 0 0 0
Kerosene 2 0 0 0
Methyl Ethyl Ketone (MEK) 3 0 – 0
Methyl Alcohol 0 0 – 0
Methylene Chloride 3 0 0 3
Naphtha 0 0 1 0
Nitric Acid 0 0 0 0
Nitric Acid (conc.) 3 0 0 –
Nitrobenzene – 0 – **
Phenol – 0 – 0
Pyridine 3 0 – –
Silver Nitrate – 0 – 0
Soap Solutions – 0 – 0
Stearic Acid – 0 – *
Sulfuric Acid 0 0 0 0
Sulfuric Acid (conc.) 3 0 0 –
Sulfurous Acid – 0 0 0
Tannic Acid – 0 0 0
Tannin Extracts – – – –
Tartaric Acid – 0 – –
Toluene 3 0 1 **
Trichloroethylene 3 0 3 3
Turpentine 2 0 0 **
Water 0 0 0 0
Xylene 3 0 0 *
* Polypropylene – satisfactory to 22°C (72°F), ** Polypropylene – satisfactory to 49°C (120°F)
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 45
4.5 Changing the measurement direction
The measurement direction can be changed by moving the whole optical
unit from above the measurement chamber to below the measurement
chamber, or vice versa.
1. Switch off the instrument and disconnect the mains power supply
cable (Fig. 3.3:2). There are anode voltages inside the mains power
supply box and the optical unit if the power is on.
2. Remove the instrument cover as described in Section 3.2 Releasing
the transportation lock.
3. Undo the four finger nuts (Fig. 4.5a:2) to release the light cover
(Fig. 4.5a:1).
4. Remove the light cover.
1
2
2
1 Light cover
2 Finger nuts, 4 pieces
Fig. 4.5a Removing the light cover of the optical unit
5. Undo the six screws, three screws on each side of the instrument
(Fig. 4.5b:1), fixing the measurement chamber to the chassis.
46 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
6. Lift the measurement chamber into the upper position from the
handle (Fig. 4.5b:2). There are hinges in the rear and a gas spring
(Fig. 4.5c:3) holds the chamber in the upper position.
Caution: The measurement chamber is very heavy. DO NOT leave
your fingers between the measurement chamber and the bottom
case.
7. Remove the three short finger nuts holding the cover (Fig. 4.5c:2).
Then remove the cover plate (Fig. 4.5c:1) from the opposite position
of the optical unit under the measurement chamber.
4
3
2
1
1 Six screws, three screws on each side of the instrument
2 Handle
3 Flat cable
4 Long finger nuts, 3 pieces
Fig. 4.5b Removing the optical unit
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 47
1
2
3
1 Cover plate
2 Short finger nuts, 3 pieces
3 Gas spring
Fig. 4.5c Cover plate
8. Lower the measurement chamber back into the down position from
the handle.
9. Unplug the flat cable (Fig. 4.5b:3) from the optical unit. DO NOT
remove any other parts from the optical unit.
10. Undo the three long finger nuts (Fig. 4.5b:4) and remove the optical
unit.
11. Lift the measurement chamber into the upper position from the
handle.
12. Place the optical unit into the position where you removed the cover
plate (Fig. 4.5c:1). Fix it with the long finger nuts and plug in the cable
connectors and the flat cable. No adjustments are needed.
13. Lower the measurement chamber back into the down position.
14. Place the cover plate instead of the removed optical unit and fix it
with the short finger nuts.
15. Fit the six screws back to hold the measurement chamber and replace
the instrument cover. The unfitted cover may increase stray light and
harm the measurement.
16. To change the measurement direction, the optical unit and the cover
plate are interchangeable.
48 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
4.6 Installing or removing the light shield
Light shield
Positioning
guide
Fig. 4.6a Installing the light shield
1. Switch off the instrument and disconnect the mains power supply cable
(Fig. 3.3:2).
2. Remove the instrument cover as described in Section 3.2 Releasing the
transportation lock.
3. Undo the four finger nuts (Fig. 4.5a:2) and remove the light cover
(Fig. 4.5a:1).
4. Undo the three long finger nuts (Fig. 4.5b:4) and remove the optical unit
or, if the optical unit is below the measurement chamber, remove the
cover plate (Fig. 4.5c:1).
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 49
5. When removing the light shield, use your little finger to lift the light
shield up. When installing the light shield, ensure that the positioning
guide is placed towards the front of the instrument against the
corresponding guide in the holder (Fig. 4.6a and Fig. 4.6b). If the light
shield is not positioned correctly, the optical unit does not slot down
into its correct place but stays swinging.
Caution: Mispositioning of the light shield will cause
discrepancies in the measurement results or may cause the
instrument not to work.
Positioning
guide
Fig. 4.6b Top view of the light shield
6. Fit the optical unit, light cover and the instrument cover back into
their places.
4.7 Installing the filters
1. Switch off the instrument and disconnect the mains power supply
cable (Fig. 3.3).
2. Remove the instrument covers as described in Section 3.2 Releasing
the transportation lock points 1 – 4.
3. If the optical unit is above the measurement chamber, undo the four
finger nuts (Fig. 4.5a:2) and remove the light cover (Fig. 4.5a:1). You
can find more detailed instructions in Section 4.5 Changing the
measurement direction.
4. If the optical unit is below the measurement chamber, undo the six
screws (Fig. 4.5b:1) fixing the measurement chamber to the chassis.
50 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
Lift the measurement chamber into the upper position. There are
hinges in the rear and a gas spring (Fig. 4.5c:3) holds the chamber in
the upper position.
Caution: The measurement chamber is very heavy. DO NOT
leave your fingers between the measurement chamber and
the bottom case.
5. Unplug the flat cable (Fig. 4.5b:3) from the optical unit. Undo the three
long finger nuts (Fig. 4.5b:4) and remove the optical unit and place it
on a table.
6. The filter wheel (Fig. 4.7a:2) can be removed by undoing the fitting
screw (Fig. 4.7a:1). DO NOT touch the filter surfaces.
7. Select the first free filter slot. DO NOT install any filters into the filter
wheel positions 7 and 8.
1
2
1 Fitting screw(s) holding the filter wheel in place
2 Filter wheel
Fig. 4.7a Filter wheels in the optical unit
8. Undo the fitting screws (Fig. 4.7b:1).
9. Remove the spring wheel (Fig. 4.7b:2).
10. Remove the spacer collar (Fig. 4.7b:3).
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 51
11. Remove the dummy filter or previously installed filter used
(Fig. 4.7b:4). Install a new filter so that the small arrow on the filter rim
points away from the filter wheel. The arrow shows the direction of
the light flow. DO NOT install any filters into the filter wheel positions
7 and 8.
12. Fit the spacer collar, spring wheel and the fitting screws. Fit the filter
wheel back into place.
4
3
13. Replace the optical unit and connect the flat cable. Lower the
measurement chamber and replace the instrument cover.
2
Fig. 4.7b Filters in a filter wheel
1
1 Fitting screw(s)
2 Spring wheel
3 Spacer collar
4 Dummy filter or previous filter used
52 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
4.8 Shutdown
• Remove any microplates left on the plate carrier or breakable strips
from the tip priming vessel. Dispose of all microplates and strips as
biohazardous waste.
• Flush the pump(s) out thoroughly with distilled water. Empty all the
tubings.
• Place the dispensing tips into the tip holder (Fig. 2.3:5).
• Switch off the Luminoskan Ascent by pressing the power switch on the
left-hand side of the instrument into the OFF position.
• Wipe the plate carrier surface and the external surfaces of the
instrument with a soft cloth or tissue paper moistened with distilled
water or a mild detergent solution.
• Push the plate carrier manually in.
• If you have spilt infectious agents on the plate carrier, disinfect with
70% pure ethanol in distilled water or some other disinfectant. See
Section 8.2 Decontamination procedure.
• If you are not using the Luminoskan Ascent for an extended period of
time, always clean the external surfaces of the instrument.
• Finally put the dust cover on.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 53
54 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
5. Maintenance
Note: Follow normal laboratory safety procedures with regard
to biohazardous, infectious, radiologic or toxic materials when
maintaining the instrument.
5.1 Routine cleaning of the instrument
For reliable operation keep the instrument free of dust and spills of liquids.
We recommend that you clean the case of the instrument periodically. A
soft cloth dampened in mild detergent is sufficient. We recommend that
you service the instrument at least yearly.
If you believe liquid has entered the luminometer, switch the instrument
off and contact your local Thermo representative or Thermo Electron or for
technical service (see Sections 8.1 Service request protocol and 8.4
Shipping the instrument (or items). If any surfaces have been
contaminated with biohazardous material, a sterilizing solution must be
used.
The prescribed decontamination procedure (see Section 8.2
Decontamination procedure), or similar routine, must be performed before
returning the instrument to the supplier for service or repair. All
instruments must be accompanied by a completed and signed Certificate
of Decontamination securely attached to the exterior of the packaging (see
Section 8.3 Certificate of Decontamination).
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 55
5.2 Cleaning the optical system
Filter wheel
Main lens
Fig. 5.2 Cleaning the optical unit
1. Switch off the instrument and disconnect the mains power supply
cable. Locate the optical unit and remove it according to the
instructions in Sections 3.2 Releasing the transportation lock and 4.5
Changing the measurement direction.
2. Clean the main lens (Fig. 5.2), the possible filters and the light shield
(Fig. 4.6a) with a cloth dampened with 96% pure ethanol and
afterwards with a lint-free cloth or a lens tissue.
DO NOT use any other liquids to clean the optical unit.
Avoid any harsh treatment.
3. Replace all the removed parts and reconnect the instrument to the
mains power supply.
5.2.1 Visual filter check
The useful life of a filter depends on environmental factors, such as dust,
humidity and temperature. Filters have a one year warranty.
Carry out the visual check in the following way:
Visually check the filter(s) by holding it (them) against an even light source.
If the color of the filter is even, then the filter is suitable for use. On the
56 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
other hand, if the filter appears to be mottled or discolored, discard the
filter since it is either damaged or defective.
The best alternative is to measure the filters with a spectrophotometer.
5.3 Cleaning the plate carrier
2
1 Plate carrier
2 A1 corner of the microplate
1
Fig. 5.3 Plate carrier
Clean the plate carrier (Fig. 5.3) with a cloth dampened with distilled water.
In case of any spills of infectious agents, clean the plate carrier with
disposable towels dampened with a disinfectant solution containing 2%
glutaraldehyde. Always use disposable gloves. Place the disposable
towels and gloves in a biohazardous waste container.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 57
5.4 Replacing the fuses
1. Switch off the instrument and disconnect the mains power supply cable
(Fig. 5.4:1). Locate the fuse holders (Fig. 5.4:2) at the rear of the
instrument.
2. Open the fuse holder to expose the fuse (Fig. 5.4:3) using a
screwdriver.
3. Replace the faulty fuse with the spare provided with the accessories or
with the same certified type.
4. Reconnect the instrument to the mains power supply. If the fuse blows
again, contact your local Thermo representative for technical service.
1
1 Mains power supply
2 Fuse holder
3
2
Fig. 5.4 Replacing the fuses
3 Fuse
58 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
5.5 Routine maintenance of optional dispensers and
main lens 1
To obtain optimum performance and maximum useful life from the
dispensers, it is important that the recommended cleaning maintenance
instructions are followed.
Luminometry is a very sensitive detection technology. Therefore, take
special care to avoid any contamination of any parts of the dispenser
tubings and follow all GLP (Good Laboratory Practice) recommendations.
5.5.1 Basic maintenance
1. The basic maintenance procedure should be performed regularly to
ensure proper dispenser operation.
2. Rinse the dispenser tubings out thoroughly with distilled water after
each use.
3. DO NOT allow the dispensers to run dry for more than a few cycles.
4. Inspect the dispensers for leaks and rectify any problems immediately.
5. Wipe up all spills on and around the dispensers immediately.
5.5.2 Extended maintenance
Clean the fluid path thoroughly using one of the procedures outlined
below. There are three ways that the dispensers may be cleaned:
• Weak detergent
• 10% bleach
• Weak acid and base
5.5.2.1 Weak detergent or 10% bleach
Remove the dispensing heads from the dispensing positions and DO NOT
let any cleaning fluids enter the measurement chamber. Use external
containers.
1. Prime the dispensers with a weak detergent or 10% bleach solution and
leave it in the dispensers with the syringes full for 30 minutes.
1
Instructions concerning the pump are reproduced from CAVRO XP 3000 Modular Digital
Pump Operators Manual made by Cavro Scientific Instruments, Inc., USA.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 59
2. After the 30-minute period, remove the aspirate tubing from the
detergent or bleach solution and remove all the fluid from the syringes
and tubing into a waste container.
3. Flush the dispenser a minimum of 10 cycles with distilled water.
5.5.2.2 Weak acid and base in sequence
Remove the dispensing heads from the dispensing positions and DO NOT
let any cleaning fluids enter the measurement chamber. Use external
containers.
1. Prime the dispensers with 0.1 M NaOH and leave the solution in the
dispensers for 10 minutes with the syringes full.
Note: DO NOT spill any 0.1 M NaOH onto any instrument
surfaces to avoid damage of the instrument. If needed, use
suitable protection covering.
2. Flush the dispensers with distilled water.
3. Prime the dispensers with 0.1 M HCl and leave the solution in the
dispensers for 10 minutes with the syringes full.
4. After the 10-minute period, remove the aspirate tubing from the 0.1 M
HCl solution and remove all the fluid from the syringes and tubing into
a waste container.
5. Flush the dispensers a minimum of 10 cycles with distilled water.
5.5.2.3 Cleaning the main lens
It is recommended that you check the main lens of the optical unit weekly.
1. Remove the optical unit if it is above the measurement chamber, or the
cover plate if the optical unit is below the measurement chamber, as
described in Section 4.5 Changing the measurement direction.
2. If needed, clean the main lens according to the instructions in Sections
5.2 Cleaning the optical system.
60 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
5.6 Periodic maintenance
There are three parts which require periodic maintenance: tubing; syringe
seals, and valves. If they become worn out, the symptoms are:
• Poor precision and accuracy
• A variable or moving air gap
• 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, the easiest and most economical way is to replace
one component at a time in the following order: (1) dispensing or aspirate
tubings and/or dispensing tip, and (2) syringe.
5.6.1 Replacing the dispenser tubings
1. To remove either the dispensing tube or the aspirate tube assembly
from the valve, gently loosen the fittings either manually or using a
7.9 mm (5/16 in.) wrench. Unscrew the fittings and remove the tubing.
2. We recommend you replace the complete assemblies always when
replacement is necessary. Alternatively, only some parts of the
assemblies can also be replaced. If only the dispensing tubing is
changed, first remove the complete dispensing tube assembly from
the dispenser unit.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 61
7
6
4
5
3
2
1
1 Dispensing head tube
2 Dispensing tip
3 Silicone tubing
4 Internal dispenser head tube
5 Brass tube holder
6 Brass locking ring
7 Dispensing tubing
Fig. 5.6.1 Structure of the complete dispensing tube assembly
3. Remove the dispensing head tube (Fig. 5.6.1:1) from the brass tube
holder (Fig. 5.6.1:5).
4. Loosen the brass locking ring (Fig. 5.6.1:6).
5. Remove the dispensing tip (Fig. 5.6.1:2) and the connecting piece of
silicone tubing (Fig. 5.6.1:3).
6. Remove the internal dispenser head tube (Fig. 5.6.1:4), the brass tube
holder and the brass locking ring.
7. Insert the new dispensing tubing into the brass tube holder/locking
ring and the internal dispenser head tube.
8. Connect the dispensing tip with the connecting piece of silicone
tubing into the new dispensing tubing.
62 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
9. Fit the dispensing tip and tubing into the dispenser head tube. When
you push the dispensing tubing gently towards the tip, the internal
dispenser head tube should be visible about 1 mm. This can be
adjusted through the length of the connecting piece of silicone tubing.
10. First screw the brass tube holder onto the dispenser head tube.
Gently tighten the tubing and ensure that the tube end stays in the
silicone tubing. Then tighten the brass locking ring.
11. To fit a new tubing, insert the fitting into the valve and tighten it
finger-tight. Using a 7.9 mm (5/16 in.) wrench, turn the fitting another
quarter to half turn.
5.6.2 Replacing the dispensing tip
3
1
2
1 Dispensing head tube
2 Dispensing tip
3 Brass tube holder
Fig. 5.6.2 Replacing the dispensing tip
1. Remove the dispensing head tube (Fig. 5.6.2:1) from the brass tube
holder (Fig. 5.6.2:3).
2. Replace the dispensing tip (Fig. 5.6.2:2) connected with a small piece of
silicone tubing in the dispensing tube.
3. Replace the dispensing head tube.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 63
5.6.3 Replacing the dispenser syringe
1
4
2 3
1 Aspirate and complete dispensing tube assemblies
2 Plunger lock screw
3 Plunger holder arm
4 Dispenser syringe (1.0 ml) and plunger
Fig. 5.6.3 Replacing the dispenser syringe
1
1. Remove the liquid from the dispenser syringe (Fig. 5.6.3:4) and from
the tubings.
2. Switch off the power from the instrument.
3. Push the plunger manually into the upper position.
4. Loosen the plunger lock screw (Fig. 5.6.3:2) approximately three full
turns.
5. Pull the plunger holder arm (Fig. 5.6.3:3) firmly down.
6. Unscrew the syringe from the valve.
7. To fit the new dispenser syringe, screw the syringe into the valve, pull
the syringe plunger down to the plunger holder arm and screw it into
place. Make sure the plunger lock screw is securely tightened.
64 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
6. Troubleshooting Guide
Note: DO NOT use the instrument if it appears that it does
not function properly.
6.1 Troubleshooting
Problem Cause Action
Instrument does not
turn on correctly
No connection between
instrument and PC
Too high background
No power connection Check the power cable
and the fuses
Incorrect software type Check that the software is
installed for the correct
instrument model
Loose serial cable
connections
Software in simulation
mode
No liquid in the well Always use some liquid in
Unclean plate Use a disposable plate
Contaminated reagents Replace the reagents
Microplate material Check if the plate
Contaminated tubing Replace/clean the tubing
Phosphorescence from
the plastic
Check the cable
connections
Switch the connection to
the instrument
the blank wells
only once
used
manufacturer or material
has changed
Use only plastics
designed for luminometry
without phosphorescence
Too low/high signal
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 65
Changed scaling factor Check the scaling factor
used
Microplate material Check if the plate
manufacturer or material
has changed Continued
Problem Cause Action
Cont.
Even, low signal level
for the whole plate
Bottom reading of nontransparent plate
Bottom reading when
drop plate in its place
Too large deviation
Unclean plate Use a disposable plate
between replicates
Dust or dirt in the wells Keep the plates protected
Foaming in the sample Use a lower dispensing or
Use correct pipetting
Illogical results Plate inserted with
incorrect orientation
Too low or variable
volume from dispenser
Inadequate priming Reprime the dispenser
Use tip priming during the
Incorrect dispensing
height
Use top reading
Remove the drop plate
only once
before use
shaking speed
techniques (reverse)
Rotate the plate with the
A1 well facing towards
the upper left corner
session
Check that the dispenser
heads are positioned at
the correct height
Incorrect dispensing
height
Check that the dispenser
head positions in the
instrument and in the
software settings are
identical
Use plate adapters with
low 384-well plates
Loose syringe, plunger
or tubings
Aspiration tube not in
the liquid
Check all the connections
in the dispenser
Ensure that there is a
sufficient volume of liquid
in the vial
Cont.
Continued
Ensure that the aspiration
66 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
Problem Cause Action
tube end weight is at the
bottom of the vial and
attach it firmly, if
necessary
Liquid droplets outside
the wells after
dispensing
Noise from the
instrument
Incorrect plate template
used
Too high total volume
used
Lid used with the plate DO NOT use plate lids
Damaged dispensing tip Replace the tip
Incorrect installation of
the dispensing head
Incorrectly adjusted
dispenser head
assembly
Light shield not
removed when using
high plates
Lid left on the plate Remove the light shield or
Dispenser head
installed to the incorrect
height
Filter wheel too tight Loosen the filter wheel
Check the plate type used
in the software
Use smaller volumes
with the dispensers
Check that the dispensing
head is inserted deep
enough
Contact authorized
technical service
Remove the light shield
the lid
Check the dispenser/plate
height
screw
Transportation lock not
removed
Plate adapter for low
384-well plates left
under a 96-well plate
Remove the
transportation lock
Remove the adapter when
using 96-well plates
Continued
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 67
Problem Cause Action
Cont.
Plate carrier moves
incorrectly
“Too high background
drift” message
Accidental dispensing
into the instrument
Foreign object, e.g., a
tip priming strip,
obstructs the plate
carrier movement
Dirty measurement
window
Contact authorized
Ensure that there are no
loose objects inside the
instrument
Clean the measurement
window
technical service
6.2 Error messages
Error messages issued by the instrument are listed in the Reference part of
the Ascent Software User’s Guide in the chapter on warning and error
messages.
68 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
7. Frequently asked questions (FAQ) about the Luminoskan Ascent
Q: Can Luminoskan Ascent be upgraded for fluorometric applications?
A: Luminoskan Ascent is a dedicated luminometer with no possibility to
add the fluorometric option. It is not possible to add the luminometric
option to the Fluoroskan Ascent. Only the Fluoroskan Ascent FL can
measure both fluorescence and luminescence.
Q: Are Petri dish adapters standard accessories that come with the
Luminoskan Ascent?
A: Petri dish adapters are not standard accessories that come with the
reader and thus they should be ordered separately.
Q: When is the adapter for low 384-well plates needed?
A: If the plate height is less than 13.5 mm, an adapter must be used to
raise the plate. The adapter for low 384-well plates is needed when you
dispense
Q: How is the scaling factor in Ascent Software used?
A: The scaling factor is a constant that multiplies all the readings from the
instrument. It is defined separately for each filter pair and beam size
(fluorometry) or filter position (luminometry). The scaling factor can be
used to convert the instrument reading to a desired level or to adjust
several instruments to give similar signal values.
and/or measure into low 384-well plates.
Q: What plate colors can be used for luminescence?
A: White plates are generally used for luminometric measurements to
enhance the signal and to avoid possible crosstalk from high signals in
neighboring wells.
Q: Can dispensing be performed into 864-well plates?
A: No, dispensing into 864-well plates is not possible.
Q: When is the light shield needed with the Luminoskan Ascent?
A: The light shield should be used with 96- and 384-well plates. If the
plates are higher than 15 mm, the light shield must be removed.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 69
Q: How many filters can be installed into the Luminoskan Ascent?
A: A maximum of six filters into the filter wheel positions 1 – 6.
Q: What plate types can be measured with the Luminoskan Ascent?
A: The Luminoskan Ascent can measure plate formats from 1 to 864.
Customized plates with maximum dimensions of
90 mm x 143 mm x 25 mm can be used.
Q: Is it possible to measure 96-well PCR plates with the Luminoskan
Ascent?
A: Yes, and you can use the Luminoskan Ascent for measuring both PCR
plates and 0.2 ml PCR tubes but you need a special adapter. This adapter
is suitable, for example, for Hybaid Omniplates, Robbins Cycloplates and
Axygen PCR microplates or tubes from Greiner or Sarstedt.
Q: What kind of plates are best to use with the Luminoskan Ascent
when you have to reduce the crosstalk as much as possible and minimize
the background?
A: There are clearly two options. You should use either white Microlite 1+
plates (Cat. no. 7571) or white universal binding plates (Cat. no. 9502887).
Q: What kind of plate adapters are available for the Luminoskan Ascent?
A: The adapters listed below are available. Other adapters are available on
request.
5210310 Adapter for Petri dish 2 x ø 59 mm
5210300 Adapter for Costar Petri dish ø 95 mm
5210380 Adapter for Falcon/Greiner Petri dish ø 93 mm
5210330 Adapter for Petri dish 6 x ø 40 mm
5210320 Adapter for low 384-well plates
5210390 Robotic 384-adapter
5210340 Adapter for Terasaki plates
5210630 Adapter for PCR tubes and 96-well PCR plates
70 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
8. Instrument Service
8.1 Service request protocol
If the Luminoskan Ascent requires service, contact your local Thermo
representative or Thermo’s service department. DO NOT under any
circumstances send the instrument for service without any prior contact. It
is imperative to indicate the fault and nature of the required service. This
will ensure a faster return of the instrument to the customer.
The Thermo representative or distributor takes care of sending Thermo’s
service department a Feedback Form (Complaint-Order), which contains a
more detailed description of the fault, symptom or condition. Give all the
necessary information to the distributor, who will fill in and forward the
Feedback Form to Thermo’s service department.
Check Section 8.4 Shipping the instrument (or items). You will find
instructions on how to proceed before shipping the instrument for service.
Check that any necessary decontamination procedure has been carried out
before packing. See Sections 8.2 Decontamination procedure and 8.3
Certificate on Decontamination. Ensure that the Certificate of
Decontamination is sent with the instrument.
The technical service department will keep you up to date with the
progress of service and provide you with any further details you might
need, for example, on maintenance, serviceability, troubleshooting and
replacement.
8.2 Decontamination procedure
Decontamination should be performed in accordance with normal
laboratory procedures. Any decontamination instructions provided with
the reagents used should be followed.
A decontamination procedure is only recommended when infectious
substances have been in direct contact with any part(s) of the instrument.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 71
If there is any risk of contamination with biohazardous material, the
procedure recommended below or some other corresponding
decontamination procedure must be performed.
We strongly recommend that the complete decontamination procedure is
performed before relocating the instrument from one laboratory to
another.
Example of disinfectants
• Formaldehyde solution 10%
• Pure ethanol 70% (in distilled water)
• Virkon solution 1 – 3%
• Glutaraldehyde solution 4%
• Chloramine T
Caution: Always use disposable gloves and protective clothing
and operate in a well-ventilated area.
Caution: DO NOT use denatured ethanol.
1. Prepare the disinfectant: 200 ml 10% formaldehyde solution or 200
ml 4% glutaraldehyde solution (or another agent recommended by
your safety officer).
2. Empty the plate carrier.
3. Switch off the power and disconnect the mains power supply cable
and the computer cable.
4. Disinfect the outside of the instrument using a cloth dampened with
70% pure ethanol in distilled water.
5. Place the instrument in a large plastic bag. Ensure that all the lids are
open.
6. Place a cloth soaked in the prepared solution into the bag. Ensure that
the cloth does not make 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.
72 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
10. Remove any stains using 70% pure ethanol in distilled water.
11. After performing this decontamination procedure, label the
instrument with a signed and dated Certificate of Decontamination.
8.3 Certificate of decontamination
The decontamination procedure is required prior to shipping the
instrument to Thermo Electron Oy, for example, for repair. If, for any
reason, the instrument is shipped back to Thermo Electron Oy, it must be
accompanied by a dated and signed Certificate of Decontamination which
must be attached to the outside of the package containing the instrument.
See Section 8.2 Decontamination procedure.
Failure to confirm decontamination will incur additional labor charges or at
worst the items will be returned for proper cleaning.
Before returning any instrument(s) or items, ensure that they are fully
decontaminated. Confirm A or B status (see below).
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 73
Certificate of Decontamination
Name: ____________________________________________________________
Address: ____________________________________________________________
Tel./Fax: ____________________________________________________________
Instrument: _________________________ Serial no.: ____________________
A) I confirm that the returned items have not been contaminated by body fluids,
toxic, carcinogenic or radioactive materials or any other hazardous materials.
B) I confirm that the returned items have been decontaminated and can be handled
without exposing the personnel to health hazards.
Materials used in the unit: Chemicals + Biological • Radioactive
*)
Specific information
about contaminants:
Decontamination
procedure:
Date and place: _________________________________________________
Signature: _________________________________________________
Name (block capitals): _________________________________________________
*) The signature of a Radiation Safety Officer is also required when the unit has
been used with radioactive materials.
This unit is certified by the undersigned to be free of radioactive contamination.
Date and place: _________________________________________________
Signature: _________________________________________________
Name (block capitals): _________________________________________________
__________________________________________________
__________________________________________________
_______________________________________________
__________________________________________________
__________________________________________________
_______________________________________________
74 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
8.4 Shipping the instrument (or items)
When you ship the instrument for service remember to:
1. Empty the dispenser(s) and remove the tube assembly.
2. Remove any loose items from the plate carrier, for example, adapters,
plates and priming vessels.
3. Remove the power cable as well as the serial cable.
4. Decontaminate the instrument beforehand.
5. Fit the transportation lock.
6. Pack the instrument according to the enclosed packing instructions.
7. Use the original packaging to ensure that no damage will occur to the
instrument during shipping. Any damage will incur additional labor
charges.
8. Inform about the use of hazardous materials.
9. Enclose a dated and signed Certificate of Decontamination (see
Section 8.3 Certificate of Decontamination) both inside and attached to
the outside of the box in which you will return your instrument or other
items.
10. Indicate the fault after you have been in touch with your local Thermo
representative or Thermo’s service department.
11. Enclose the return authorization number (RGA) given by the Thermo
representative.
See Section 14.1 General specifications for details on storage and
transportation temperatures.
8.5 Service contracts
We strongly recommend that you maintain and service the instrument
every twelve months on a contract basis by the manufacturer’s trained
service engineers. This will ensure that the product is properly maintained
and gives trouble-free service. Contact authorized technical service for
further details.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 75
76 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
9. Disposal of the Instrument and Materials
9.1 Disposal of the instrument
For the disposal of the instrument do the following:
• Decontaminate the instrument prior to disposal. See
Sections 8.2 Decontamination procedure and 8.3
Certificate of Decontamination.
• 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.
• Regarding the original packaging and packing
materials, use the recycling operators known to you.
• For further information, contact your local Thermo
representative.
9.2 Disposal of materials
Refer to local regulations for the disposal of infectious material.
The samples can be potentially infectious. Dispose of all used
microplates, PCR tubes, disposable gloves, syringes,
disposable tips, etc., as biohazardous waste.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 77
78 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
10. Ordering Information
10.1 Product code numbers
Microplate Luminometers
Item Cat. no.
Luminoskan Ascent 5300160
Luminoskan Ascent with dispenser 5300170
Additional dispenser option (factory installation) 5210230
Accessories
Item Cat. no.
1st Dispenser kit (user installation) 2805621
Additional dispenser kit (user installation) 2805630
Plate carrier kit 2806140
Adapter for Costar Petri dish ∅ 95 mm 5210300
Adapter for Falcon/Greiner Petri dish ∅ 93 mm 5210380
Adapter for Petri dish 2 x ∅ 59 mm 5210310
Adapter for dispensing into a low 384-well plate 5210320
Adapter for Petri dish 6 x ∅ 40 mm 5210330
Adapter for Terasaki plate 5210340
Drop plate 2805880
Light shield 1006470
Dispensing tip 1047661
Complete dispensing tube assembly
Aspirate tube 24071510
Dispensing tubing (Fig. 5.6.1:7) 24071500
Dispensing syringe (1.0 ml) and plunger (Fig. 3.4:4) 24071490
Syringe seal 2805600
3-port valve 2805610
Leakage tray (Fig. 2.3:4) 10478030
End weight 1002380
Red adjustment collar (Fig. 4.4.4b:2) 1002820
Silicone tube (for connecting pieces) 0412290
RS-232C interface cable D9 Female/D25 Female 2305290
RS-232C interface cable D25 Female/D25 Female 2303760
Fuse 3.5 A/250 V UL 198G Time Delay (10 pcs min. order) 1210950
Dust cover 1610460
Luminoskan Ascent User Manual 1507520
Ascent Software User’s Guide 1507530CD
Thermo Electron Ascent Software 5185450CD
(Fig. 3.4.2 and Fig. 5.6.1) 24071580
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 79
Thermo Electron Microtiter plates
Cat. no. Description Bottom Qty
Thermo Microtiter Plates for Luminescence
7416 Microlite 1 Plate Flat 50/box
7417 Microlite 2 Plate Flat 50/box
7418 Microlite TCT Plate, sterile
7571 Microlite 1+ Plate Flat 50/box
7572 Microlite 2+ Plate Flat 50/box
7521 Microlite 1+ Plate U 50/box
7522 Microlite 2+ Plate U 50/box
9502887 White 96 Well Plate UB Flat 50/box
95029580 White 96 Well Plate EB Flat 50/box
95029770 White 96 Well Plate Sterile Flat 40/box
Cat. no. Description Bottom Qty
Thermo Microtiter Plates for Luminescence, Strips and Assemblies
7421 Microlite 1 1x12 Strip assembled Flat 100/box
7403 Microlite 1 1x12 Strip Flat 320/box
7400 Microlite 2 1x12 Strip assembled Flat 100/box
7410 Microlite 2 1x12 Strip Flat 320/box
7561 Microlite 1+ 1x12 Strip assembled Flat 100/box
7566 Microlite 1+ 1x12 Strip Flat 320/box
7562 Microlite 2+ 1x12 Strip assembled Flat 100/box
7567 Microlite 2+ 1x12 Strip Flat 320/box
95029510 White Strip 1x8 assembled UB Flat 50/box
95029530 White Strip 1x12 assembled UB Flat 50/box
95029660 White Breakable Strip 1x8 Assembled UB Flat 50/box
Cat. no. Description Qty
Thermo Microtiter 384 Well Plates
95040010 White 384 Round Well Plate 50/box
95040230 White 384 Round Well Plate Sterile with lid 40/box
50/box
FFllaatt
80 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
Thermo Electron Microtiter plates Cont.
Cat. no. Description Bottom Qty
Thermo Microtiter Streptavidin-Coated Plates
95029273 White BioBind Strip 1x8 Assembled Flat 5/box
95029303 White BioBind Br. Strip 1x8 Assembled Flat 5/box
Cat. no. Description Qty
Thermo Microtiter Plate Accessories
1)
5500
Universal Polystyrene Lid, Sterile 100/box
1)
5550
Styrene Individually Wrapped Lid, Sterile
9503210
9503220
6305 Vinyl Lid for 1x12 strip assembly 100/box
6604
6000
1)
Accessories for ex-Dynex products
2)
Accessories for ex-Thermo products
2)
Microplate Lid, 96-well 15/box
2)
Microplate Lid 15/box
1)
Holder for 1x12 strip assembly 10/box
1)
Workstation 1x12 strip assembly 1/box
50/box
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 81
10.2 List of recommended spare parts
Cat. no. Item 1–2 unit(s)/year 10 units/year
24071580 Complete dispensing tube
assembly
24071500 Dispensing tubing 2 8
1047661 Dispensing tip 4 20
2805690 Aspirate tube assembly 1 5
24071490 Dispenser syringe
(1.0 ml) and plunger
1210950 Fuse 3.5 A/250 V UL 198G
Time Delay
1006470 Light shield 1 3
1002820 Red adjustment collar 2 10
1 5
1 5
1 5
10.3 Ordering filters
When ordering luminometric filters, please contact Thermo Electron Oy.
E-mail address: info.microplateinstruments@thermo.com.
82 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
11. Glossary and Abbreviations
ATP Adenosine triphosphate, a biological molecule
that is commonly used as a reference chemical
for luminometric sensitivity.
Chassis The framework of the instrument.
Chemiluminescence A compound that emits light following a
chemical reaction is said to be
chemiluminescent.
Crosstalk Interfering signal from neighboring wells.
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.
Disinfection The destruction of pathogenic bacteria, usually
with an antiseptic chemical or disinfectant.
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
7
.
Error message Indication that an error has been detected.
Initialization Initialization tests are so-called self-tests, which
are carried out prior to operation to ascertain
that the necessary instrument adjustments have
been carried out.
LED Light-emitting diode.
Luminescence Emission of light (other than from thermal
energy causes) such as bioluminescence.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 83
Luminometer
An instrument used for measuring the intensity
of luminescent radiation.
Luminometric label
(Luminophore)
A substance which emits light at room
temperature. A group of atoms that can make a
compound luminescent.
Photomultiplier tube
(PMT)
A photoelectric cell that converts light into
electric current and amplifies the current.
RH Relative humidity.
RLU or rlu Relative Luminescence/Luminometric/Light
Units. The arbitrary units in which luminescence
intensity is reported.
rpm Revolutions per minute.
Scaling The measured values are expressed as RLU.
Scaling is a way to convert readings to show
desired values.
Self-tests Initialization tests and adjustments that the
instrument performs prior to operation as well
as autocalibration.
84 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
12. Literature
Fluorescent and Luminescent Probes for Biological Activity. A Practical
Guide to Technology for Quantitative Real-Time Analysis (1999). Mason W.
T., (ed.). Second edition, Biological Techniques Series, Academic Press.
Bioluminescence and chemiluminescence - Methods in Enzymology, Vol.
57 (1978). DeLuca M. A. (ed.). Academic Press., New York; 653 pages.
Bioluminescence and chemiluminescence: Part B - Methods in
Enzymology, Vol. 133 (1986). DeLuca M. A. and McElroy W. D. (eds.).
Academic Press, Orlando; 649 pages.
Chemiluminescence: Principles and Applications in Biology and Medicine
(1988). Campbell, A. K.. Horwood, Chichester, U.K.; 608 pages.
Nonradioactive labeling and detection of biomolecules (1992). Kessler C.
(ed.). Springer-Verlag, Berlin; 436 pages.
Bioluminescence Methods and Protocols (1998). LaRossa R. A. (ed.).
Methods in Molecular Biology, Vol. 102. Humana Press., Totowa, New
Jersey; 310 pages.
Bioluminescence and chemiluminescence: Perspectives for the 21
st
century (1999). Roda A., Pazzagli M., Kricka L. J., Stanley P. E. (eds.). John
Wiley & Sons, Chichester, U.K.; 625 pages.
Bioluminescence and Chemiluminescence: Part C - Methods in
Enzymology, Vol. 305. (2000). Ziegler M. M. and Baldwin T. O. (eds.).
Academic Press, San Diego; 732 pages.
Bioluminescence & Chemiluminescence (2001). Case J. F., Herring P. J.,
Robison B. H., Haddock S. H. D., Kricka L. J., Stanley P. E. (eds.). World
Scientific Publishing Co. Pte. Ltd., Singapore; 517 pages.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 85
86 Luminoskan Ascent
User Manual Rev. 2.1, Cat. no. 1507520
13. Warranty Certificate
Thermo Electron Microplate Instrumentation Business products are fully
guaranteed against defective parts and materials, including defects caused
by poor workmanship, for a period of one year from the date of delivery.
Thermo will repair or replace defective parts or materials during the term
of warranty at no extra charge for materials and labor provided that the
products were used and maintained in accordance with Thermo’s
instructions. The warranty is invalid if products have been misused or
abused.
For the warranty to be effective, the product must have been purchased
either directly from Thermo or from an authorized Thermo distributor. The
guarantee is not transferable to a third party without prior written approval
from Thermo.
This guarantee is subject to the following exclusions:
• Any defects caused by normal wear and tear.
• Defects caused by fire, lightning, flood, earthquake, explosion,
sabotage, war, riot, or any other occurrence of the type listed
above.
• Refurbished products that are subject to different warranty
conditions.
THIS WARRANTY IS IN LIEU OF ALL OTHER EXPRESSED OR IMPLIED
WARRANTIES, INCLUDING BUT NOT LIMITED TO ANY IMPLIED
WARRANTIES OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR
PURPOSE. The seller is not liable for any loss or damage arising out of or
in connection with the use of the product or other indirect damages.
These warranty terms and conditions can be obtained from your local
Thermo dealer.
This card acts as a warranty certificate.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 87
13.1 Warranty limitations
The following items are not included in the warranty:
• Consumables
• Software programs
• Fuses.
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14. Specifications
Thermo Electron reserves the right to change any specifications without
prior notice as part of our continuous product development program.
14.1 General specifications
Weight
Basic unit 21 kg (46 lbs.).
3 optional dispensers add 3.5 kg to the weight.
Overall dimensions 420 mm (16.5 in.) (W) x 420 mm (16.5 in.) (D) x
340 mm (13.4 in.) (H), options included.
Operating conditions
(indoor use)
Transportation conditions
Storage conditions
Mains power supply 100 – 240 Vac, 50/60 Hz, nominal
Fuses 2 x 3.5 A/250 V,
Power consumption 200 VA max., 32 VA standby.
Computer interface Serial RS-232C port. Baud rate 9600. Character
+10°C – +40°C, RH 90% max.
Tested according to IEC 60068-2-1 test Ab, (Cold).
IEC 60068-2-2 test Bb, (Dry heat).
IEC 60068-2-3 test Ca, (Damp heat).
-40°C – +70°C, packed in transportation packaging.
Tested according to IEC 60068-2-1 test Ab, (Cold).
IEC 60068-2-2 test Bb, (Dry heat).
-25°C – +50°C, packed in transportation packaging.
Tested according to IEC 60068-2-1 test Ab, (Cold).
IEC 60068-2-2 test Bb, (Dry heat).
UL 198G Time Delay, 5 x 20 mm.
format 1 start bit, 8 data bits, 1 stop bit, no parity.
Flow control XON/XOFF.
Continued
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 89
General specifications Cont.
Detector Photomultiplier tube.
Filters for luminometric
measurements
Plate types 1- to 384-well plates. Can also be programmed for
Shaker Built-in orbital shaker with adjustable speed and
Incubator Temperature range from RT (25°C) +3°C to 45°C,
Dispensers 1 – 3 optional dispensers. Syringe volumes 1000 µl.
Filters can be used; maximum 6 filters.
nonstandard configurations. Maximum dimensions
90 mm x 134 mm x 25 mm.
diameter.
when the ambient temperature is 25°C. The
temperature is selected using Ascent Software.
Dead volume 600 µl. Metal-free fluid path.
Exchangeable valve, syringe, tubing and tip.
Autoclavable tubing and tip.
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14.2 Safety specifications
The Luminoskan Ascent fulfills the following requirements:
EN 61010-1:1993 + A2:1995/IEC 61010-1:1990 + A1:1992 + A2:1995
Installation Category (Overvoltage Category) II; Pollution Degree 2
EN 61010-2-010:1994 + A2:1996/IEC 61010-2-010:1992 + A1:1996
CSA C22.2 No. 1010.1 M1992
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
Mains supply
+5°C – +40°C
± 10% (if larger than specified above)
fluctuations
Installation category
II according to IEC 60664-1 (see Note 1)
(overvoltage category)
Pollution degree 2 according to IEC 60664-1 (see Note 2)
Notes
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 II 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.
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.
Both of these affect the size of the electrical dimensioning within the
instrument.
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 91
14.3 In conformity with the requirements
The Luminoskan Ascent bears the following markings:
Type 392
100 – 240 Vac 50/60 Hz, 200 VA
CE mark
CSA monogram
UL monogram
The Luminoskan Ascent conforms to the following requirements:
73/23/EEC (Low Voltage Directive)
89/336/EEC (Electromagnetic Compatibility Directive, EMC)
FCC Part 15, Subpart B/Class B
Safety performance:
EN 61010-1:1993 + A2:1995/IEC 61010-1:1990 + A1:1992 + A2:1995
EMC performance:
EN 50081-1:1992 Generic emission standard. Residential,
commercial and light industry.
EN 50082-1:1997 Generic immunity standard. Residential,
commercial and light industry.
EN 61326-1:1997 + A1:1998 Product family standard.
Test standards
EN 55022:1998 Class B, 150 kHz – 1 GHz
EN 61000-3-2:1995 + A1:1998
+ A2:1998 + A13:1997 +
A14:2000
EN 61000-3-3:1995
ANSI C63.4:1992 Class B, 450 kHz – 1 GHz; 30 MHz – 1000 MHz
EN 61000-4-2:1995 + A1:1998 4 kV CD, 8 kV AD, Criteria B
EN 61000-4-3:1996 + A1:1998 3 V/m, 80 MHz – 2 GHz, Criteria A
EN 61000-4-4:1995 1 kV, Criteria B
EN 61000-4-5:1995 2 kV line to ground, 1 kV line to line, Criteria B
EN 61000-4-6:1996 3 V
EN 61000-4-8:1994 3 A/m, Criteria A
EN 61000-4-11:1994 30%/10 ms, Criteria B
60%/100 ms, Criteria C
> 95%/5 s, Criteria C
Performance limits
Class A
, 150 kHz – 80 MHz, Criteria A
rms
100%/20 ms, Criteria B
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14.4 Performance specifications
Warm-up time < 15 min to rated accuracy.
Measuring speed Depends on the plate type and the measurement
type. The minimum kinetic interval time is 15 s for a
96-well plate (from well A1 back to the same well
A1).
Luminometric spectral
270 – 670 nm
range
Luminometric
Up to approx. 5000 Relative Light Units (RLU)
measurement range
Luminometric sensitivity 1 fmol ATP/well (typical) in a white Thermo Electron
96-well strip plate
(Thermo Electron ATP monitoring kit).
Luminometric dynamic
Limit = (2 x SD
> 9 decades over whole gain setting area
Blank
) / I
Ref-Blank
x C
Ref
range
Shaker Orbital method, speed 60 – 1200 rpm,
∅ 1 – 50 mm
Incubator
Heaters:
Warm-up time from RT (25°C) to 37°C, 15 min.
Liquid in the well:
Ambient temperature 23°C, covered 96-well plate,
200 µl/well.
Temperature accuracy:
Mean temperature of the wells ± 1°C
Uniformity: ≤ 1°C
Warm-up speed 1 h from 23°C to 37°C on an
average 90% of the set value and the ambient
temperature.
Dispensers Dispensing volume 1 – 1000 µl in 1 µl increments.
Accuracy ± 3 µl avg. for volumes 5 µl and above
Precision 5 – 19 µl < 5%
20 – 1000 µl < 2%
Minimum dispensing speed 25 s, 96-well plate,
5 µl/well, tested with distilled water.
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15. Index
A
Abbreviations....................................................................................... 3, 83
Accessories.......................................................... 19, 20, 25, 58, 69, 79, 81
Adapter..................................................... 10, 26, 28, 29, 42, 67, 69, 70, 79
Application ................................................................. 7, 8, 9, 10, 43, 69, 85
B
Blanking.............................................................................................. 30, 33
Blanks ........................................................................................... 34, 65, 99
Bottom reading ................................................................ 10, 26, 33, 45, 66
Brass tube holder............................................................................... 62, 63
Brief User’s Guide................................................................................ 3, 99
C
Chassis ................................................................................... 13, 46, 50, 83
Chemical resistance................................................................................. 43
Chemiluminescence .......................................................................... 83, 85
Clean optical system ............................................................................... 56
Consumables ........................................................................................... 87
Control switches .................................................................... 15, 16, 39, 40
Cover sensor............................................................................................ 15
Crosstalk............................................................... 10, 11, 16, 33, 69, 70, 83
D
Decontamination ................................................. 55, 71, 73, 74, 75, 77, 83
Certificate of ........................................................... 55, 71, 73, 74, 75, 77
procedure................................................................ 53, 55, 71, 72, 73, 77
Detector.................................................................................................... 90
Dimensions ............................................................................ 42, 70, 89, 90
Directives ................................................................................................. 92
Dispenser
head tube.................................................................................. 42, 62, 63
syringe ............................................................................................ 64, 82
tubing............................................................................ 16, 39, 59, 61, 99
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 95
Dispensing head .......................... 15, 17, 25, 38, 39, 40, 59, 60, 62, 63, 67
holder........................................................................................ 15, 25, 38
Dispensing tip ................................................ 43, 53, 61, 62, 63, 67, 79, 82
Dispensing tubing ............................................................ 61, 62, 63, 79, 82
Disposal of materials ............................................................................... 77
Drop plate................................................................... 26, 27, 29, 39, 66, 79
Dummy filter ............................................................................................ 52
Dummy plug....................................................................................... 15, 38
E
End weight ................................................................................... 26, 67, 79
Environmental requirements............................................................. 21, 99
Error message.............................................................................. 31, 68, 83
F
FAQ....................................................................................................... 3, 69
Feedback Form ........................................................................................ 71
Filter
check..................................................................................................... 56
order...................................................................................................... 82
wheel................................................................. 16, 33, 51, 52, 67, 70, 90
Frequently asked questions ................................................................ 3, 69
Fuses..................................................................... 14, 58, 65, 79, 82, 87, 89
G
Gain........................................................................................................... 93
Glossary................................................................................................ 3, 83
I
Incubator ............................................................................ 9, 17, 38, 90, 93
Initialization ............................................................................ 30, 31, 83, 84
Instrument layout..................................................................................... 13
Intended use............................................................................................. 10
L
Leakage tray ....................................................................................... 15, 79
LED ............................................................................................... 30, 31, 83
Locking piece ..................................................................................... 22, 23
Locking ring........................................................................................ 62, 63
Luminescence .............................................................. 9, 10, 69, 80, 83, 84
Luminometric label .................................................................................. 84
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Luminophore ........................................................................................... 84
M
Maintenance ................................................................ 3, 55, 59, 61, 71, 75
N
Noise .................................................................................................. 21, 67
O
Operational check.................................................................................... 30
Ordering information............................................................................... 79
P
Packing ............................................................................. 19, 20, 71, 75, 77
instructions..................................................................................... 19, 75
list.................................................................................................... 19, 20
materials ......................................................................................... 19, 77
PCR ............................................................................................... 28, 70, 77
Photomultiplier tube.................................................. 10, 11, 16, 33, 84, 90
Plate adapter .............................................................. 27, 28, 29, 66, 67, 70
Plunger ....................................................................... 25, 26, 64, 66, 79, 82
Positioning lever ................................................................................ 18, 32
Prime Tip.................................................................................................. 40
R
Replace......................................... 20, 48, 52, 56, 58, 61, 63, 65, 67, 71, 87
fuses...................................................................................................... 58
RLU ............................................................................................... 34, 84, 93
S
Scaling.................................................................................... 34, 65, 69, 84
factor......................................................................................... 34, 65, 69
Self-tests ...................................................................................... 10, 83, 84
Service ....................................................... 3, 19, 30, 55, 58, 67, 68, 71, 75
request protocol............................................................................. 55, 71
Shaker .......................................................................... 9, 10, 35, 66, 90, 93
Shutdown....................................................... 46, 49, 50, 53, 56, 58, 64, 72
Signal........................................................................................................ 65
Spacer collar ...................................................................................... 51, 52
Luminoskan Ascent User Manual Rev. 2.1, Cat. no. 1507520 97
Specifications..................................................................... 3, 75, 89, 90, 91
general ...................................................................................... 75, 89, 90
performance ......................................................................................... 93
safety..................................................................................................... 91
Spring wheel ...................................................................................... 51, 52
Switching on ................................................................................ 24, 31, 53
T
Tip priming ................................................................. 18, 40, 41, 53, 66, 68
Top reading ........................................................................................ 33, 66
Transport .................................................................................................. 20
Transportation lock............................ 20, 22, 23, 46, 49, 50, 56, 67, 75, 99
Troubleshooting............................................................................. 3, 65, 71
guide ..................................................................................................... 65
U
Unpacking .......................................................................................... 19, 99
W
Warnings ...................................................................... 7, 21, 24, 29, 31, 68
Warranty................................................................................... 3, 19, 56, 87
98 Luminoskan Ascent
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Appendix A. Brief User’s Guide
• Unpack the instrument (p. 19).
• Check delivery for completeness (p. 20).
• Check for damage during transport (p. 20).
• Place the instrument according to environmental requirements
(p. 21).
• Release the transportation lock (p. 22).
• Install the dispenser tubings if equipped with dispensers (p. 25).
• Plug in the instrument (p. 23).
• Switch the instrument ON and ensure that the green light is lit
(p. 31).
• Install Ascent Software (p. 31).
• Create an Ascent Software session (see Ascent Software User’s
Guide: Procedure Desktop|Session|New).
• We recommend you perform a dummy run first with an empty
plate when you have created a new session and then a run with
known samples or controls.
• Load the microplate with prepared samples, blanks and controls.
Place the microplate onto the plate carrier of the instrument so
that the A1 well is located in the upper left corner of the plate
carrier (p. 32).
• Start the session by pressing the START button on the
Procedure Desktop toolbar of Ascent Software.
• Follow the instructions provided by the software.
• Switch the instrument OFF after routine operation (p. 53).
• Maintain the Luminoskan Ascent instrument on a regular basis
(p. 55).
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