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User manual
1420
Software version 3.00
1420-922-07
March 2003
PerkinElmer Life and Analytical Sciences, Wallac Oy, P.O. Box 10, FIN-20101 Turku, Finland.
Tel: 358-2-2678111. Fax: 358-2-2678357. Website: www.perkinelmer.com/lifesciences
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Contents
Contents
Introduction .......................................................................................................................... 9
System overview............................................................................................................... 11
Enhanced Security mode .................................................................................................. 12
Software installation ......................................................................................................... 13
Operation overview .......................................................................................................... 14
Operating Wallac 1420....................................................................................................... 15
Starting up Wallac 1420 ................................................................................................... 17
Without enhanced security mode.................................................................................. 17
With enhanced security mode....................................................................................... 17
Instrument control............................................................................................................. 18
Live display ...................................................................................................................... 19
Temperature...................................................................................................................... 21
Barcode options ................................................................................................................ 22
Plate orientation................................................................................................................ 22
Starting operation with the Start button............................................................................ 23
Starting operation with the Start Wizard .......................................................................... 26
Starting operation and using a protocol number barcode ................................................. 28
Stacker operation .............................................................................................................. 29
Starting operation, no protocol number barcode .......................................................... 29
Starting operation and using a protocol number barcode ............................................. 31
Ending operation............................................................................................................... 32
Stop button.................................................................................................................... 32
End button..................................................................................................................... 32
Button on Wallac 1420 ..................................................................................................... 33
Light on the button ....................................................................................................... 33
Protocol editing ................................................................................................................... 35
Explorer ............................................................................................................................ 37
Explorer folders ............................................................................................................ 38
Explorer icons............................................................................................................... 38
Protocol selection.......................................................................................................... 39
Protocol editor .................................................................................................................. 40
Samples............................................................................................................................. 41
ID...................................................................................................................................... 43
Measurement .................................................................................................................... 44
Measurement mode....................................................................................................... 44
Measurement operations............................................................................................... 45
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Down button ............................................................................................................. 45
Up button ..................................................................................................................45
Delete button............................................................................................................. 45
Dispense button......................................................................................................... 45
Delay button.............................................................................................................. 46
Shake button ............................................................................................................. 46
Label button .............................................................................................................. 46
Kinetics button .......................................................................................................... 47
Scanning button ........................................................................................................ 47
Injector position during measurement....................................................................... 47
Plate .................................................................................................................................. 48
Plate repeat parameters .................................................................................................48
Measurement height...................................................................................................... 48
Plate type....................................................................................................................... 49
Temperature level checking .......................................................................................... 49
Outputs.............................................................................................................................. 50
Events................................................................................................................................ 52
Keywords ...................................................................................................................... 53
General.............................................................................................................................. 54
File menu ..........................................................................................................................55
Save............................................................................................................................... 55
Print............................................................................................................................... 55
Print preview................................................................................................................. 55
Print setup .....................................................................................................................55
Start............................................................................................................................... 55
Exit................................................................................................................................ 55
Tools menu ....................................................................................................................... 56
Result viewing ..................................................................................................................... 57
Latest results .....................................................................................................................59
Results in the Explorer...................................................................................................... 59
List ................................................................................................................................ 61
Plate map.......................................................................................................................62
Protocol......................................................................................................................... 63
Notes ............................................................................................................................. 64
Error .............................................................................................................................. 64
Print............................................................................................................................... 64
Export results ................................................................................................................ 64
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Tools menu .......................................................................................................................... 65
Tools menu ....................................................................................................................... 67
User level.......................................................................................................................... 68
Labels ............................................................................................................................... 69
Time-resolved fluorometry........................................................................................... 71
Protocol name ........................................................................................................... 71
Flash Energy Area .................................................................................................... 72
Flash Energy Level ................................................................................................... 72
Excitation Filter ........................................................................................................ 72
Light Integration Capacitors ..................................................................................... 72
Note .......................................................................................................................... 73
Light Integration Reference Level............................................................................ 73
Emission filters ......................................................................................................... 73
Emission aperture ..................................................................................................... 73
Counting parameters................................................................................................. 74
Factory-set values for the main counting window.................................................... 74
Beam size.................................................................................................................. 74
Second measurement parameters.............................................................................. 74
Fluorometry .................................................................................................................. 75
Protocol name ........................................................................................................... 75
CW-Lamp energy ..................................................................................................... 76
CW-Lamp Control Mode.......................................................................................... 76
CW-Lamp filters....................................................................................................... 76
Excitation aperture (Wallac 1420-040/041/042/043 only) ....................................... 76
Emission filters ......................................................................................................... 76
Emission aperture ..................................................................................................... 77
Counter position ....................................................................................................... 77
Counting time ........................................................................................................... 77
Second measurement parameters.............................................................................. 77
Photometry.................................................................................................................... 78
Protocol name ........................................................................................................... 78
Absorbance mode ..................................................................................................... 78
Flash Lamp Filter...................................................................................................... 78
CW-Lamp filters....................................................................................................... 79
Excitation aperture (Wallac 1420-040/041 only) ..................................................... 79
Reading time............................................................................................................. 79
Second measurement ................................................................................................ 79
Luminometry ................................................................................................................ 80
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Protocol name ...........................................................................................................80
Emission filters .........................................................................................................80
Emission aperture...................................................................................................... 81
Counting time............................................................................................................ 81
Second Measurement ................................................................................................ 81
Fluorescence polarization (Wallac 1420-040/041/042/043 only)................................. 82
Protocol name ...........................................................................................................82
CW-Lamp energy...................................................................................................... 83
CW-Lamp filter......................................................................................................... 83
Polarizer aperture ...................................................................................................... 83
Emission filter........................................................................................................... 83
Emission aperture...................................................................................................... 83
Counting time............................................................................................................ 83
G factor .....................................................................................................................84
LANCE ......................................................................................................................... 85
Protocol name ...........................................................................................................85
Flash Energy Area..................................................................................................... 86
Flash Energy Level ................................................................................................... 86
Excitation filter .........................................................................................................86
Light integration capacitors ...................................................................................... 86
Note........................................................................................................................... 87
Light Integration Reference Level ............................................................................ 87
Emission filter........................................................................................................... 87
Emission aperture...................................................................................................... 87
Counting parameters ................................................................................................. 88
Factory-set values for the main counting window .................................................... 88
Label .........................................................................................................................88
Delay 1...................................................................................................................... 88
Delay 2...................................................................................................................... 88
Window 1.................................................................................................................. 88
Window 2.................................................................................................................. 88
Cycle ......................................................................................................................... 88
Beam size .................................................................................................................. 88
Second measurement parameters .............................................................................. 88
LANCE Label Properties.......................................................................................... 89
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Filters................................................................................................................................ 90
Emission filters ............................................................................................................. 91
CW-Lamp filters........................................................................................................... 92
Filter slides ...................................................................................................................93
Emission filter slide .................................................................................................. 94
Physically changing the emission filter slide............................................................ 95
CW-Lamp filter wheel.............................................................................................. 96
Physically changing the CW-Lamp filter wheel....................................................... 97
Changing the CW-Lamp........................................................................................... 97
Changing filters in the software................................................................................ 98
Filter properties......................................................................................................... 99
EuSm Dual label normalization...................................................................................... 100
Plate type selection ..................................................................................................... 100
Preparing the plate ...................................................................................................... 101
Measurement status page............................................................................................ 102
Measurement warning page........................................................................................ 102
Normalization confirmation page ............................................................................... 103
LANCE normalization.................................................................................................... 104
Using the LANCE normalization wizard.................................................................... 104
Protocol selection........................................................................................................ 105
Preparing the plate ...................................................................................................... 106
Measurement status page............................................................................................ 107
Measurement warning page........................................................................................ 108
Normalization confirmation page ............................................................................... 109
Using LANCE normalization samples in the assay.................................................... 110
Plate dimension wizard................................................................................................... 112
Plate selection ............................................................................................................. 113
Technology selection .................................................................................................. 113
Preparing the plate ...................................................................................................... 114
Define the centre point of the corner wells................................................................. 115
Miscellaneous settings .................................................................................................... 117
Plate type .................................................................................................................... 119
Plate name............................................................................................................... 119
Number of well rows .............................................................................................. 120
Number of well columns......................................................................................... 120
Height of the plate .................................................................................................. 120
Strip orientation ...................................................................................................... 120
Plate dimensions ..................................................................................................... 120
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Tune ........................................................................................................................ 120
Well names.................................................................................................................. 120
Instrument properties .................................................................................................. 120
Options............................................................................................................................ 121
General........................................................................................................................ 121
Instrument hardware ...................................................................................................124
Measurement technologies.......................................................................................... 125
Exiting................................................................................................................................ 127
File menu ........................................................................................................................129
Automatic logout ........................................................................................................129
Troubleshooting ................................................................................................................ 131
Out of memory................................................................................................................ 133
Display problems ............................................................................................................ 134
Performance degradation ................................................................................................ 134
Processing error ..............................................................................................................135
System requirements not met.......................................................................................... 135
Appendices......................................................................................................................... 137
Appendix 1: Connecting MultiCalc to Wallac 1420....................................................... 139
Introduction................................................................................................................. 139
Wallac 1420 MultiCalc assay protocols...................................................................... 139
Installation of Wallac 1420 output option in MultiCalc .............................................140
MultiCalc assay protocols........................................................................................... 140
Usage of MultiCalc ..................................................................................................... 140
Appendix 2: Plate types for stackers............................................................................... 141
Wallac ..................................................................................................................... 141
Nunc........................................................................................................................ 141
Costar ...................................................................................................................... 141
Greiner ....................................................................................................................141
Appendix 3: Dispenser operation.................................................................................... 143
Introduction................................................................................................................. 143
Emptying the waste vial.............................................................................................. 144
Preparing the tubing.................................................................................................... 146
Dispenser maintenance ...............................................................................................147
Measurement............................................................................................................... 150
Maintenance................................................................................................................ 151
Needle maintenance ................................................................................................ 151
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Appendix 4: Enhanced security special functions .......................................................... 153
User groups and permissions...................................................................................... 153
User Management....................................................................................................... 154
Password Control........................................................................................................ 156
Audit Trail .................................................................................................................. 158
Archive ....................................................................................................................... 160
Security Settings ......................................................................................................... 161
Index .................................................................................................................................. 165
Trademarks
DELFIA is a registered trademark and LANCE is a trademark of PerkinElmer, Inc.
Windows and Excel are registered trademarks of Microsoft Corp. in the U.S. and other
countries.
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Introduction
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Introduction
System overview
Note: This manual describes all the features of Wallac 1420. Some versions of the
instrument do not have one or more of these features. You can ignore the parts of this
manual not relevant to the version of Wallac 1420 you have.
Wallac 1420 multilabel counter is a complete platform for quantitative detection of lightemitting, or light absorbing markers. Depending on the model of the instrument you have, it
is suitable for flash or glow luminometry, fluorometry, fluorescence polarization, highsensitivity time-resolved fluorometry, homogeneous time-resolved fluorometry (LANCE
option) and photometry. It is a compact bench top unit with features such as dispensers,
shaking, temperature control, top or bottom reading and scanning. The software is a 32-bit
application running under Windows
counted (depending on the technology and model) as well as Petri dishes, slide filters and
Terasaki plates. Plates can be loaded manually or with stackers for automatic operation.
Output can be to a file on the PC and/or to a laser printer. The following figure shows the
Wallac 1420 system with its PC and laser printer.
2000 or Windows XP. Up to 1536-well plates can be
If the dispenser unit is connected to Wallac 1420 it looks as follows:
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Introduction
Enhanced Security mode
Wallac 1420 has an Enhanced Security mode intended for facilities that have to comply
with 21 CFR Part 11 regulation from the Food and Drug Administration (FDA) of the USA.
During installation you can select if you want to use the Enhanced Security mode. Once you
have enabled this mode you cannot disable it because to do so would not be in compliance
with 21 CFR Part 11.
The regulation (21 CFR Part 11) sets the criteria under which the agency (FDA) considers
Electronic Records and Electronic Signatures (if applied) to be trustworthy, reliable and
generally equivalent to traditional paper records and hand-written signatures executed on
paper. The regulation applies to records in electronic form that are created, modified,
maintained, archived, retrieved, or transmitted, under any records requirements set forth in
agency regulations (i.e. GMP, GLP, GCP…). It also applies to electronic records submitted
to FDA under requirements of the Federal Food, Drug, and Cosmetic Act.
The Enhanced Security mode for Wallac 1420 provides the technological controls and
features to support full 21 CFR Part 11 compliancy. These features can be classified under
three main headings:
• improved access control (unique Userid/Password combination) with five user
levels
• improved data security
• audit trails of user actions
This manual and the electronic help will tell you how to use Wallac 1420 with or without
the Enhanced Security mode. All differences in use are noted and explained.
Note: use of the Enhanced Security mode does not alone ensure compliance with 21
CFR Part 11. Any facility that wants to be compliant with the 21CFR Part 11
regulation must also implement the necessary procedures and controls set by the
regulation.
Please refer to Appendix 4 for details of special functions that are part of the Enhanced
Security mode.
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Introduction
Software installation
Note: the following instructions assume that the counter, PC and printer have been
connected up and switched on (see the installation instructions in the Instrument manual). If
Enhanced Security mode is used then installation can only be done by a person with System
and Windows administrator rights. See Appendix 4 for more details.
1. Load the software CD.
Load CD
2. The setup loader will start automatically and give you a choice of either selecting the
Installation Wizard to guide you through the installation or reading files from the CD.
3. When you have completed the installation you should remove the CD from the PC,
otherwise the setup loader will start again next time you boot the PC.
Note: in the case of Windows XP the user has to have “power user” rights.
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Introduction
Operation overview
Parameters must first be set using the protocol editor. This is activated by selecting a
protocol in the Explorer.
A sample plate can then be loaded and measured using the protocol set for it.
Other parameters affecting the whole system e.g. labels, filters etc. can be set in the Tools
menu.
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Operating Wallac 1420
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Starting up Wallac 1420
Without enhanced security mode
When you boot the system the following window appears:
Operating Wallac 1420
With enhanced security mode
When the Enhanced Security mode is installed the login dialogue appears after you have
started Wallac 1420.
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Operating Wallac 1420
Type the appropriate userid/password combination to proceed. Clicking the Cancel button
closes Wallac 1420. Clicking the Change password opens the Change Password dialogue
where you can define a new password.
After a successful login Wallac 1420 can be used according to the permissions granted to
the logged on user.
Instrument control
The main window has three tabs: Instrument Control, Live Display and Temperature. The
instrument control tab enables you to control the instrument by starting, stopping or ending
counting. It shows you the protocol being used to control the measurement and it also shows
status of the instrument.
In the enhanced security mode these controls are enabled only for users who belong to the
Operator user group.
If the system supports plate heating, the status bar will display the current temperature of the
plate and samples. When run with a system not supporting plate heating this will not be
displayed.
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Operating Wallac 1420
When the workstation is used without an instrument (i.e. running in demo mode), the status
bar includes the text “DEMO”. When Wallac 1420 workstation is connected to an
instrument (running in normal mode), the demo mode indicator is blank. You can switch
between normal and demo modes using the Options selection in the Tools menu.
Live display
The Live display shows you the live results in the form of a plate map with colour coding
to indicate the size of the results. If you move the cursor on to a well that has been
measured, the numerical value will appear. At the bottom of the window there is
information about the sample being measured including its current value.
Note: in multiple label, scanning and kinetics measurements, only results from the first
measurement of the label are visible in the Live display. You can see results from all labels
in the Latest results display which you can get by clicking the Latest assay run icon.
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Operating Wallac 1420
You can choose the result range you want used to determine the colour code of the wells.
The closer a result is to the upper limit of the range, the more red it will appear. The closer
to the lower limit it is, the more blue it will appear. These colours will correspond to the
values shown in the legend appearing beside the plate map. Choose the range that is
appropriate for the samples you are measuring so that you get the greatest variation in
sample colours.
You can choose whether you want the scale to be logarithmic (which means that the
difference between smaller values will be more visible whereas the difference between
higher values will be less visible), or a linear scale in which every part of the range is
treated equally.
In the enhanced security mode these controls are enabled only for users who belong to the
Operator user group.
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Temperature
Operating Wallac 1420
In the Temperature tab of the main window you select plate heating and the target
temperature. The latter can be set with 0.1
minimum temperature is 15
o
C or room temperature plus 2
o
C precision within the interval 15 – 45o C. The
o
C depending on which is
higher. If the target is changed, the new value is only effective when the Apply button has
been clicked. The figure shows the temperature rising rapidly to its new level and then
stabilizing there. If the plate heating option is not installed, these controls are disabled.
The current temperature is shown in the bottom right hand corner of the window
irrespective of which tab is selected.
If the plate heating option is not installed, these controls are disabled.
In the enhanced security mode these controls are enabled only for users who belong to the
Operator user group.
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Operating Wallac 1420
Barcode options
An optional barcode reader can be included. This allows you to load barcode labelled plates,
which are identified by the barcode reader. Codabar, Code39, Interleaved 2 of 5, Code 128,
UPC and EAN barcodes can be read.
There are two sorts of identification that can be used:
This identification can be by plate number, in which case you must select the protocol as
part of the start up procedure.
Alternatively, the barcode can be the number of the protocol to be used for measuring that
plate. This system is especially useful for models using stackers. In this case the first plate
to be loaded from the stacker must have a barcode. Measurement can be started simply by
briefly pressing the button on the counter. See page 118 for more details on barcode usage.
Plate orientation
The figure shows a typical 96-well plate and how it is to be orientated when loading it either
directly into Wallac 1420 or into an input stacker. The A1 position must be in the left corner
furthest from you i.e. the side that enters the counter first. If a barcode is used, it should also
be on the side that enters the counter first. The barcode can then be used to identify the plate
or to select the protocol.
Optional barcode
Loading
direction
Position
A1
Plates used in a stacker
must have this ledge at
least on the short sides
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Operating Wallac 1420
Starting operation with the Start button
1. Open the loading position lid.
2. Load your first plate and close the lid. See the picture on page 20 for correct plate
orientation.
Load first plate
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Operating Wallac 1420
3. Select the protocol you want to use by scrolling through the list of active protocols.
4. Use the mouse to click the Start button on the user interface
You can follow the measurement process by selecting the Live display.
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Operating Wallac 1420
5. When the plate has been measured, remove it and load the next plate.
1. Remove the
measured plate
2. Load the
next plate
3. Press the
button briefly
6. Briefly press the button on the counter, this will be blinking if there are more plates to be
loaded for this assay. If it is not blinking then the assay has finished.
Blinking light
shows assay
Continue until all plates have been measured.
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Operating Wallac 1420
Starting operation with the Start Wizard
In the enhanced security mode the Start Wizard is enabled only for users who belong to the
Operator user group.
An alternative way to start counting is to use the Start Wizard. This allows you to define
how many plates you are going to use and to give the positions of samples. Proceed as
follows:
1. Click on the Start Wizard icon and follow the instructions that appear.
The Start Wizard will appear to guide you through the procedure for starting.
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The steps are:
Introduction to the wizard. (You can select the introduction not to appear next time if you
want).
Protocol selection - click the protocol you want to use for measurement.
Definition of the sample names and number of plates. See Samples tab on page 39 for a
description of the principles involved in sample definition.
Note: Errors in protocol selection or plate layout definition will lead to incorrect results.
Comments that you want added to the results - just type in what you want
Load the first plate - the protocol name and comments appear in order to confirm that it is
the one you want. Note: the A1 position must be in the left corner furthest from the user i.e.
the side that enters the counter first. If a barcode is used it should also be on the side that
enters the counter first.
Click Finish to start measurement. See the Live display during measurement.
When the plate has been measured, remove it, load the next plate and briefly press the
button on the counter.
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Operating Wallac 1420
Starting operation and using a protocol number barcode
1. When you load a plate, the button will be blinking. See the picture on page 20 for correct
plate orientation.
Load the first
plate
Light is
blinking
2. Press the button briefly. Operation will start. The button light will be lit without blinking.
The barcode on the plate will be read and the protocol specified by the barcode will be
selected and used for measuring the plate.
When the measurement has finished and the light starts blinking again, remove the plate and
load the next one and briefly click the button. If the plate belongs to the same assay as the
previous one, no barcode is needed. If it is barcoded for a new assay then the appropriate
protocol will be used for measuring the plate.
1. Remove the
measured plate
2. Load the
next plate
Continue measuring plates this way. Note that the button light will remain blinking as long
as the protocol number barcode option is selected.
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Operating Wallac 1420
Stacker operation
A stacker is able to hold up to 20 (or 40 in the larger version) microplates. No lids, loose
frames or very loose strips are acceptable. See Appendix 2 for a list of plates that fit in the
stackers.
Starting operation, no protocol number barcode
Caution Do not put your fingers into the sample loading area. In the stacker
model the loading area is uncovered when the stackers are not in place so you
must avoid the danger of getting your fingers trapped by the plate lift mechanism.
If you need to do something in that area,
1. In stacker operation, plates are loaded into the input stacker with the first plate to be
measured at the bottom. See the picture on page 20 for correct plate orientation.
2. The empty output stacker (the one with the tabs for the quick release mechanism) should
be fitted first into the multilabel counter; it occupies the left-hand position.
3. Fit the input stacker (with plates) in the right hand position.
switch power off first.
1. Load
the output
stacker
Release tab for the stacker
Note: Make sure the stackers have clicked into position properly.
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2. Load
the input
stacker
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Operating Wallac 1420
p
Stackers are
loaded ready
for operation
Quick release
mechanism for
lates
4. Select the protocol and click the Start button or use the Start Wizard.
5. The plates will be moved one by one from the input stacker to the measurement position.
After each plate has been measured, it is moved to the output stacker. When the assay has
been completed operation will stop.
If there are still plates in the input stacker it means that you must start a new assay by
selecting the protocol and clicking
6. When all plates have been moved to the output stacker, it can be removed. To do this,
press the release tab at the bottom of the stacker, then lift it up.
Start as described above.
After you have removed the stacker, pull up the handles of the quick release mechanism to
allow the plates to be emptied easily from the stacker.
this mechanism when the stacker is loaded in Wallac 1420 because you may jam the
conveyor.
If the button on Wallac 1420 is blinking, it means that the assay has not finished and you
must remove the empty input stacker, load an empty output stacker and then an input
stacker which has the remaining plates of the assay (plus plates for other assays if you
want).
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Note: make sure you do not operate
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Operating Wallac 1420
Starting operation and using a protocol number barcode
Caution Do not put your fingers into the sample loading area. In the stacker
model the loading area is uncovered when the stackers are not in place so you
must avoid the danger of getting your fingers trapped by the plate lift mechanism.
If you need to do something in that area,
This configuration makes best use of the stackers; operation is automatic.
1. Load the stackers. The button light will be blinking.
switch power off first.
Light is blinking
2. Click the button briefly.
One by one all the plates will be loaded and measured. If a plate has a barcode, then it will
be measured using the protocol specified by the barcode. If it has no barcode, it will be
measured using the protocol specified by the previously read barcode.
3. Only when the input stacker is empty will operation stop. The button light will be
blinking. Remove the stackers and load new ones if required.
4. Click the button briefly to start operation again.
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Operating Wallac 1420
Ending operation
Stop button
End button
Clicking the End button in the workstation ends the assay after the
current plate has been completely measured, including any possible
repeat measurements. The plate is brought out as described above.
Clicking the Stop button in the workstation interrupts the
measuring process immediately. The plate being measured is
brought out to the loading position. You cannot resume
measurement after clicking
the beginning if required.
Stop but must begin the assay from
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Operating Wallac 1420
Button on Wallac 1420
This button has different functions depending on how long you press it for and what is
happening when you press it.
If a plate has been measured, you can remove it, load a new plate and briefly press this
button to continue the assay with the same protocol as the plate just measured. However, if
you press this button for more than 2 s, the current assay will be stopped and a new assay
will be started with the current protocol. This will not work if the protocol barcode option is
selected.
If a plate is being measured and you press this button, nothing happens.
If no assay is running and you press the button for more than 2 s, then a new assay will be
started with the current protocol. This will not work if the protocol barcode option is
selected or the enhanced security mode is installed.
Note: make sure in all cases that you have loaded the correct plate before pressing the
button because the plate in the loading position (or the empty plate holder) will be taken
into the counter.
Light on the button
The following table shows you what the light on the button will be like depending on what
is happening with Wallac 1420 and whether or not the protocol barcode option is used.
Condition Barcode none or plate
number
Not measuring, no assay Off Blinking
Not measuring, waiting next
plate
Measuring, loading or
unloading
Blinking Blinking
On On
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Barcode for protocol
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Operating Wallac 1420
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Protocol editing
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Protocol editing
Explorer
In the enhanced security mode Explorer is enabled only for users who belong to the Editor
user group.
Through the Explorer you can access the Protocol Editor and the Result Viewer. Click the
Explorer icon to open the Explorer.
The Explorer will open.
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Protocol editing
Explorer folders
In the Wallac folder there are, as a default, folders named according to the technologies
installed. In each of these folders there are protocols using the appropriate technology.
On the same level as the Wallac folder there are two other folders: Users and Projects.
To open a folder, click on it and then select Open from the File menu. Alternatively you
can open a folder by double-clicking the left mouse button when the cursor is on the folder
icon. You can also click once on the + mark next to the folder to see the protocols contained
in it.
To close a folder, either double-click on it or click the - sign next to it.
If you are working on the Advanced or Service level, you can create, rename or delete
folders. A sub folder of Users or Projects can be renamed by selecting it and clicking the
right mouse button and selecting
Rename.
Explorer icons
Factory pre-set protocols and groups have a lock symbol to show they cannot be changed.
User made protocols and groups do not include the lock because the user can change them.
See the examples below:
The icon for a factory pre-set folder
The icon for a factory pre-set protocol
The icon for a user-made folder
The icon for a user-made protocol
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Protocol editing
Protocol selection
You can: Open, copy, paste, start or create a protocol, delete a user made protocol (plus
its associated results). In the enhanced security mode you have to belong to the Operator
user group to start a protocol.
To open an existing protocol you can do one of the following:
a) Move the mouse cursor onto it and then click the left mouse button to select it, then click
the
File menu and click Open.
b) Move the cursor on to it, click the right mouse button and select Open from the menu
that appears.
c) Double-click the mouse when the cursor is on the icon.
At the bottom of the window on the status line the protocol ID will appear (if defined,
otherwise it is not available, N/A) and also any notes that have been included in the
protocol.
When you create a protocol you can save it e.g. as a user protocol in the Users folder, or
you can save it according to the technology. The choice is yours, but the default structure
gives you a starting point and helps you to find the pre-set protocols.
You can move a user made protocol to a different folder by selecting it, holding down the
mouse button, dragging it to the target folder and releasing the mouse button.
Note: The dual label protocol (only available if TR-FIA is installed) should not be copied
and edited e.g. to make a single label europium or samarium protocol. You should begin
with an existing single label protocol. For dual label operation, you must select the
europium label first and then the samarium label. After you have selected the second label, a
third, samarium in the europium window, will be created automatically. When running with
these three labels you must have a valid Eu/Sm normalization, see the
to make such a normalization.
For information about results see the chapter on "Result viewing".
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Protocol editing
Protocol editor
Note: changes to protocols should only be made by authorized persons.
The protocol editor window contains seven tabs:
Samples - shows a plate map to allow you to edit wells and plates
ID - you can identify the protocol
Measurement - you can select the labels used for measurement
Plate - you can select the plate type and other parameters
Outputs - you can define the output
Events - you can give a command to be followed after an assay has been run with this
protocol
General - information about the protocol
You can set parameters for up to 40 plates in one protocol.
Protocol entries are checked for validity by the software and invalid entries are informed
when you try to switch from one view to another.
When you have edited a protocol you can use the command line or the button bar to save or
print the protocol.
Note - if you want to change the plate type, do this before setting other plate parameters.
This is because when the plate type is changed all the previous plates will be deleted and
one plate of the selected type will appear.
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Samples
Protocol editing
Selecting this tab causes a plate map to be displayed to allow you to edit samples. Buttons
allow you to select the
You can also add a new plate, delete the currently selected plate or duplicate a plate. An
added plate has all the positions marked as Measured and will become the last plate in the
sequence. When you delete a plate it is the one currently visible that is deleted. A duplicated
plate is a copy of the current plate and will become the last plate in the sequence.
A map of all the wells appears. An empty well (i.e. one that will not be measured) will be
grey, a well with a sample (i.e. one that will be measured) will be coloured. If you move the
cursor onto a well the sample type will appear. If you click on the well a list of available
well types will appear.
first plate, previous plate, next plate or last plate.
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Protocol editing
To change the markings for the individual wells in a plate you can select a group of wells by
using the mouse. To start the selection, click the left mouse button at the desired starting
position and then drag the mouse cursor to the desired ending position on the plate without
releasing the left mouse button.
Complete rows or columns can be selected by clicking on the character/number on the
plate frame corresponding to the row/column. More than one row/column can be selected
by clicking the mouse and dragging in the same way as when selecting free well areas.
The whole plate can be selected by clicking on the upper left corner of the plate frame.
To select a rectangular area of wells on the plate, hold down the Shift key while dragging
with the mouse.
The number of wells selected is shown in the plate indicator text after the plate order
number.
When you have selected the wells and released the left mouse button, a drop down menu of
sample types will appear. These will be the types defined in the Wallac 1420 Manager
Tools menu item Miscellaneous settings. By selecting one of these types you specify what
the type of the selected area is to be.
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ID
Protocol editing
This tab allows you to select the protocol number and name. The number can be selected
from a drop down list of available numbers. The protocol name has to be typed in. In the
case of a factory pre-set protocol you cannot change the number.
Notes or additional comments can be connected with the protocol. These appear at the
bottom of the Explorer window.
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Protocol editing
Measurement
The measurement tab specifies the measurement sequence to be used in the protocol.
Note: If the protocol being edited is pre-set, the measurement sequence cannot be changed.
Measurement mode
This field allows you to specify on what unit the complete measurement sequence should be
performed before moving on to the next similar unit. The possible unit is: a single well, a
strip or a plate.
Plate and strips modes are disabled when using a fast kinetic or scan measurement in the
operation sequence. Well mode will be disabled when using more than one label in the
operation sequence. However, copying and pasting of the same label is possible. Plate mode
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Protocol editing
can be used for slow kinetics by making repeated counting of the plate with delays in
between.
Measurement operations
This field lists the operations that have been specified to be used in the assay. The
operations are listed in the order they will be performed.
If you select an item in the list and then click the right mouse button, a menu will appear to
allow you to
beside the Operations list enables you to add items to the list. Click the button referring to
the operation you want. Set the parameters and click
Down button
This moves the selected operation in the operation list one step closer to the end. This
button is enabled when the operation currently selected in the measurement operations list is
not the last operation in the list.
Up button
This moves the selected operation in the operation list one step closer to the beginning. This
button is enabled when the operation currently selected in the measurement operations list is
not the first operation in the list.
Delete button
This deletes the operation selected in the operation list. This button is enabled when there is
an operation selected in the measurement operations list.
Copy, Delete or see the Properties of the selected item. A row of buttons
OK to add the item to the list.
Dispense button
Clicking this causes a dialogue to appear that lets you specify the parameters for a dispense
operation. This button is enabled when the dimensions of the plate type used in the protocol
are 8x12 or less. You can select
the number of injectors to be used (from 1 to 4 depending on how many are
installed)
the volume to be dispensed (in microlitres). A slide bar helps you to select this
volume, 350 microlitres is the maximum
the speed from 1 to 5 (where 5 is the fastest and 4 the default)
the increment (using a slide bar with a range from plus 100 microlitres to minus
100 microlitres), this is the volume added to or subtracted from the previous
volume dispensed before the next dispensing
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Protocol editing
the number of replicates (from 1 to 3)
the injection mode. You can either aspirate and then dispense the same amount
(aspVol=dispVol) or you can fill the whole syringe and then dispense several times
(aspVol=syringeVol). The former is more accurate but the latter is faster.
At the bottom of the dialogue is a check box “Perform on first plate repeat only”. If you
only want dispensing for the first of the plate repeats, check this box. If you want
dispensing for every repeat then leave it unchecked. Clicking the
OK button inserts a
dispense operation in the measurement operations list.
Delay button
Clicking this causes a dialogue to appear that lets you specify the duration of the delay
between the completion of the preceding operation and the start of the one succeeding the
delay operation in the operation list. You use a slide bar to do this. Clicking the
OK button
inserts the delay operation into the measurement operations list. A single delay can be from
0.1s to 3600 s.
At the bottom of the dialogue is a check box “Perform on first plate repeat only”. If you
only want delay for the first of the plate repeats, check this box. If you want delay for every
repeat then leave it unchecked.
Shake button
Clicking this causes a dialogue to appear that lets you specify the parameters for a shake
operation. You can specify the duration of the operation, the speed (slow, normal or fast),
the extent the shaker moves and the path that it follows - straight line, elliptical or figure of
eight (linear, orbital or double orbital respectively). Clicking the
OK button inserts the
shake operation into the measurement operations list.
At the bottom of the dialogue is a check box “Perform on first plate repeat only”. If you
only want shaking for the first of the plate repeats, check this box. If you want shaking for
every repeat then leave it unchecked.
Label button
Opens the Label Selector to let you specify a label to be inserted in the measurement
operations list. There can be up to 10 labels in the list. Note: for LANCE measurements two
labels must be selected. If only one is selected then the measurement will be treated as
normal time-resolved fluorescence.
When using well or strips mode, the label button will be disabled when there is already a
label in the measurement sequence. However, copying and pasting of the same label is
possible. For detailed information about labels see Labels on page 67.
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Kinetics button
Clicking this causes a dialogue to appear that lets you specify the parameters for a kinetic
measurement. You can give the number of repeat measurements (up to 100) and the delay
between each measurement (0 to 600 s). You need to also select the label by clicking the
label button and then selecting from the label dialogue.
If the repeat is less than 26 and the delay between repeats is 0, results will come in one
group. The kinetics button is enabled only when using well mode and there are no other
labels in the measurement sequence. However, copying and pasting of the same kinetic
label is possible.
Scanning button
Clicking this causes a dialogue to appear that lets you specify the parameters for a scan
measurement. This means that several measurements are made for a single well and the
measurements are made from different points of the well. These points are in the form of an
array for which you can give the number of horizontal steps, the number of vertical steps
and the point displacement i.e. the distance between each point on the well where the
measurement is made. The total number of points can be a maximum of 100. The
distribution of points can be square or rounded.
You also need to select the label by clicking the Label button and then selecting from the
label dialogue.
The scanning button is enabled only when using well mode and there are no other labels in
the measurement sequence. However, copying and pasting of the same scanning label is
possible.
Injector position during measurement
At the bottom of the Measurement tab are two check boxes for controlling the injector head.
These will only be enabled if you have selected the “by wells “ mode You can select if you
want the measuring head to be kept above the well during measurement. This means that
measurement can be started immediately after injection without waiting for the injector head
to move aside.
If you check this box then a second check box is enabled where you can select if you want
injecting and measurement to happen simultaneously. This is for use in very fast kinetic
measurements.
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Protocol editing
Plate
Plate repeat parameters
The number of times each plate is to be measured (1-99). If this is more than 1 then you
must give the time between the end of one measurement and the beginning of the next (03600 secs.). This allows you to do slow kinetic measurements.
Measurement height
Standard - This selection is only enabled when using a plate type that has its dimension
data stored in the instrument.
The default height set is about 8 mm above the bottom of the plate.
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User defined - If you do not want to use the standard measurement, you can give a new
value here. You must click the “
mm if the dispenser is used) and the maximum 18.0 mm depending on the plate type and if
the dispenser is used or not. The height is measured from the bottom of the plate. This is
useful if you are measuring samples on filter paper and you need to optimise for the sample
being at the top or at the bottom of the wells.
user defined” button. The minimum value is 3.0 mm (8.0
Plate type
Select the one you want from the list of available plate types under the scroll bar. If the plate
size and/or format is different from the current one all the plates in the plate map will be
deleted and one plate of the selected type will appear. Make sure you select the plate type
before defining the plate map to avoid losing your settings.
Plate types are read from the Wallac 1420 database when the Protocol Editor is started so it
will not recognize plate types added while the Protocol Editor is open.
Note: the generic 8 x 12 plate selection does not check the height of the plate before a
measurement.
Note: If the type you selected has a different number of wells than the current one, all the
previous plates will be deleted and one plate of the selected type will appear.
Temperature level checking
This setting allows you to define what the target temperature should be when a plate is
measured. The plate heating controls are only enabled when used with a system that
supports the plate heating option.
Note: Setting this option will not activate the plate heating automatically, but it will cause a
warning to appear if the temperature differs significantly from what has been specified in
the protocol.
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Protocol editing
Outputs
The Outputs tab in the Protocol editor allows you to select where results will be sent at the
end of a run. This can be to a printer and/or file. Click the check box of the output device(s)
you want.
If you have selected Printer output, you must select the format, either plate format (results
are in a table format with rows and columns in the order shown in the plate map) and/or list
format (results are in a single column starting with those from the first well). When this
option is not set, the “Include in output” checked list box is disabled. This list box defines
which parts of the assay run data should be included in the printed output: List, Plate,
Protocol description, Error description, Notes.
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Protocol editing
If you have selected File output you must specify the file type from the drop down menu of
possible types. The types you can save files in are Excel 5 or later versions, Tab delimited
text or MultiCalc.
If the Include in output list box is checked, then it specifies which optional parts of the
assay run information should be included in the file saved when the measurement is
completed. These are the same as for printer output: List, Plate, Protocol description, Error
description, Notes.
You must also type in the file name. This can include keywords connected with the file as
described in Events. The button to the right of the filename box gives you a list of all valid
placeholder arguments to choose from. Other filenames possibly used in other protocols are
also available through this button.
Note: you will not be allowed to change to another tab if an invalid placeholder argument is
found. However, if a previous file with the same file name exists, it will be overwritten by
the new file without warning.
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Protocol editing
Events
This tab allows you to give a command that will be executed when a protocol has been run.
The keyword should be in triangular brackets < > exactly as the examples given in the tab
show. These commands are then automatically replaced by the appropriate name or number
etc. after the assay run ends.
The first box specifies the executable to run, while the second box specifies the commandline arguments to use. The button to the right of the first box lets you browse to find the
executable file you want to use. The button to the right of the second box again lists all the
valid keyword arguments that you can choose from, see Keywords below.
For example, you can start an external spreadsheet running and have it execute the file
produced by the run. This file will be identified by e.g. protocol name or number. There are
also unique internal identifiers that the system gives for each protocol and assay run.
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Protocol editing
Keywords
The following keywords can be used for file names in the protocol editor Outputs tab and in
Assay end system command in the Events tab:
<ProtocolName>, the name of the protocol. Specified in the ID tab in the protocol name edit
box.
<ProtocolNumber>, the protocol number for the protocol. Specified on the ID tab in the
protocol number box.
<ProtocolID>, a unique numeric identifier for the protocol.
<AssayID>, a unique numeric identifier for the assay run.
<RunID>, a numeric value identifying the specific run of a certain protocol.
<RunDate>, date when the assay was run in the format YYYYMMDD.
<Plate barcode>, the barcode of the first plate of the assay. If there is no barcode on a plate,
you should define an explicit file name for the plate.
<Output file name>, the name of the output file given in the Outputs tab.
In some cases the numeric value of the AssayID or the RunID may not be suitable for the
program handling the file. You can add a function “pad” to allow you to decrease or
increase the number. You can specify <AssayIDpadX> where X=1..7 or <RunIDpadX>
where X=1..4.
Examples
If <AssayID> = 1234 and you add pad 2 to get <AssayIDpad2> the result will be 34. This
would be useful with MultiCalc which could not accept an assay ID as large as 1234.
If <AssayID> = 234 and you add pad 4 to <AssayIDpad4> the result would be 0234. This
would be useful if you needed a four digits identifier.
The validity of possible keyword arguments is checked when you try to switch from this
view to another. If an invalid argument is found you are notified and the page switch is
cancelled.
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Protocol editing
General
This tab gives you information about the protocol, when it was created, when it was edited
most recently, by whom it was created and edited and when an assay was last run with it.
The user names used come from the identifier given when Windows was booted. Pre-set
protocols are created in the factory.
In the enhanced security mode the first user name is the system user name and the second is
the Wallac 1420 user name. User names are separated with a slash.
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Protocol editing
File menu
The protocol editor file menu contains the following items of which Save and Print have
corresponding buttons.
Save
When you select this item or click the button the current protocol information is saved.
When you select this item or click the button a dialogue opens up for you to select the
printer type and other details of the printout.
Print preview
This item allows you to see what will be printed out. You can select whether you want a
side by side two page view or a single page view. You can also zoom in (magnify the page)
to see more details, alternatively you can zoom out (reduce the magnification) if the page is
already magnified. You can step to the next page or back to the previous one. When you
have finished viewing the pages that are going to be printed out you can click the
button to get the printout or Close to return to the protocol editor.
Print setup
You can define the printer type that will be selected. You can also set the paper size and
orientation.
Start
Starts measurement with the currently selected protocol.
Print
In the enhanced security mode you must belong to the Operator user group to start the
measurement.
Note: if the protocol has been changed you will be prompted to save the protocol before
measurement is started.
Exit
This closes the protocol editor. If the protocol has been changed it will prompt you to save it
first.
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Protocol editing
Tools menu
The Tools command line menu has one item Options. When you click this a dialogue with
a check box appears. If you do
measurement of the selected protocol will begin directly. If you do check it, then, whenever
you click the
selected protocol.
In the enhanced security mode the Options menu item is enabled only for users who belong
to the Service user group.
Start icon, the Start Wizard will be used to start the measurement of the
not check it, then, whenever you click the Start icon,
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Result viewing
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Result viewing
Latest results
The results from the latest assay can be viewed by clicking the Latest assay run icon.
In the enhanced security mode you must belong to the Operator user group to view the
results of the latest assay.
The Result Viewer is started and the latest results shown after the measurement has been
completed. This command performs following actions:
1. If there are no assay runs in the database yet, a warning is given and the Result
Viewer is not invoked.
2. If this button is clicked while an assay run is already in progress, that assay is
displayed in the Viewer.
3. If this button is clicked and there is no current assay running, the latest assay run is
displayed on the Viewer.
Results in the Explorer
Alternatively, once results have been received and stored in the computer, you can view
them by means of the Wallac 1420 Explorer. See the picture on the next page.
The protocol connected to the latest results will be selected automatically, but you can select
any protocol that was used to make the measurements that you want to view. The results
files associated with the protocol will then appear on the right side of the window. Each
entry for a results file comprises the following items Assay ID, the date and time when the
assay began and when it ended, Notes and Errors.
Note: to change a column width in the Results Viewer, move the mouse cursor to the line
between two columns. The cursor form will change to a vertical bar with two arrows. Hold
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Result viewing
down the left mouse button and move the column division in the direction you choose. The
information in a column is complete if no “.....” are shown at the end.
Click on the result file and then click the Open icon.
Note: these results are in the form of raw numerical data
The Result Viewer gives details of the actual measurement. There are five tabs and these
described below.
The Result Viewer also provides you the following functions by means of command line
menus or, in most cases, buttons:
File - Export (Button), Page setup, Print (Button) and Exit.
View - List (Button), Plate (Button), Protocol (Button), Errors(Button), Notes (Button),
View all result data (only when List is selected), Toolbar, Status bar
Plate - first plate, previous plate, next plate, last plate (all with corresponding buttons).
These buttons or menu items that allow you to step between plates only appear when the
plate mode is selected.
Help - information about Result Viewer.
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Result viewing
List
The following information appears: Plate ID, Number of repeats, Well number and Sample
type abbreviation (e.g. M= Measured). In addition there are pairs of columns. The number
of pairs depends on what you have set in protocol editing parameters. The first column of
each pair is the measurement time and the other the result value. You can change column
widths to get more information. In the case of high-density plates results can be split over
several tabs.
View all result data - This item in the View menu is only active if List view is selected. It
causes an extra column to appear in the result list giving the following extra information:
Time-resolved fluorometry –number of flashes
Fluorescence – measurement time
Luminometry – counting time
Photometry – A/D converter value and the number of readings
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Result viewing
Plate map
This plate map shows you the results arranged from top to bottom in the order they
appeared in the list from left to right. See List format for the alternative way of viewing
results.
The cells in the two first rows displayed include information about the currently displayed
plate. The information included is:
Plate: The number of the physical plate currently being displayed.
Repeat: The number of the repeat of the physical plate currently being displayed.
Measurement end time: Date and time when the measurement of the result plate was
finished.
Start temperature: Plate holder temperature when the measurement of the plate was
started.
End temperature: Plate holder temperature when the measurement of the plate ended.
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Result viewing
Barcode: Barcode of the plate if used, if not used it displays the text ‘N/A’.
The cells displayed in the rows directly above each result plate gives information directly
relating to the label or result type displayed in the plate grid below it. The information
included is:
Name of the label or result type displayed in the result plate
Unit for the results from the result plate
Instrument Background value measured for the label or result type.
Protocol
Selecting Protocol causes a list of protocol parameters to appear. There is also a plate map
showing which sample types were on the plate. Any notes written in the protocol editor
Notes box will appear here.
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Notes
This tab allows you to see any notes that are connected with the results being viewed.
Error
Error gives you information about the general error situation and specific information about
any errors that occurred for each measurement type.
Clicking this icon leads to an extended print dialogue which in addition to the normal print
options gives you a choice of printing
is the default. You can also click the
printed.
In the case of high-density plates the results from the first columns (1 to 18 or 1 to 24)
appear on one page and the rest (19-36 or 25 to 48) appear on the next page. Only rows of
one plate will be printed on one page.
If you press OK in the print dialogue, the selected parts of the run information will be
printed on the printer that has been selected in the print dialogue.
selected sheets or the entire workbook. The former
Preview button to get a preview of the pages to be
Export results
If you select Export in Result Viewer it allows you to export the results of the active plate.
A window appears in which you can give the path, the file name (type it in) the file type,
(select it from the list), a
Save button and a Cancel button.
The file types you can save files in are Excel 5.0 or later, Tab delimited text or MultiCalc .
To help you select where you want the file to be saved, you can browse to find the folder
you want. Buttons allow you to go up one level, create a new folder, and display the folder
information as a list just showing folder names or with additional details. The white area in
the centre of the window is where the folders appear when you start to browse to find the
path.
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Tools menu
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Tools menu
Tools menu
Note: These operations can affect the whole system and should only be undertaken by
someone qualified to do them.
There are up to fifteen items in this menu (depending on the options installed):
Explorer- for protocol editing and result viewing, see pages 35 and 57.
Start Wizard - for starting when you first need to define plates and wells, see page 24.
Results of latest assay run - the result viewer is started and the latest results shown, see
page 57.
Labels – for defining labels used in measurements.
Filters – for defining filter types.
EuSm dual label normalization wizard, - (requires TR-FIA to be installed).
LANCE normalization wizard – (requires LANCE to be installed).
Plate dimension wizard – for fine-tuning the plate position when measuring high density
plates
Dispenser maintenance – described in Appendix 3.
Miscellaneous settings – including plate types and well names.
User level – selecting the features accessible to the user.
Users – (appears only in the enhanced security mode). The “User level” menu item is
replaced with this one.
Audit Trail – (appears only in the enhanced security mode).
Archive – (appears only in the enhanced security mode).
Security – (appears only in the enhanced security mode).
Options – includes demo mode selection, barcode and plate map features.
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Tools menu
User level
Note: in the enhanced security mode extra menu items appear.
The User level in the Tools menu allows you to select the user level: Routine, Advanced or
Service.
If the User Level is selected to be Routine, this will automatically disable any system
operations. To enable these operations you must select the
for qualified service specialists. It gives access to e.g. Instrument Options.
Advanced level. Service is only
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Tools menu
Labels
In the enhanced security mode this tool is enabled only for users who belong to the Editor
user group.
In this window there are tabs corresponding to the measurement technologies (labels) in the
instrument. Each tab includes the names of the currently defined labels. Names of user
defined labels can be freely defined by the user but names of factory pre-set labels cannot
be changed. See the sections below for details.
There are four buttons at the bottom of the labels window:
Add - add a new item to the list. Give the name and edit the properties.
Copy - (select an item to activate this). Make a copy of an item. It will appear with a new
icon showing it is different from a default item, and the name will be preceded by the words
“Copy of”. You can give it a different name and edit other parameters by selecting
Properties.
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Tools menu
Remove - (select a user created item to activate this). Remove the selected item from the
list. Preset items and labels used in protocols cannot be removed. This command requires
confirmation because you cannot undo Remove.
Properties - (select an item to activate this). View the properties of the selected item. If the
item is user defined then you can edit the properties, but if it is a factory pre-set, you cannot
edit them.
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Tools menu
Time-resolved fluorometry
Note: to be enabled this requires the TR-FIA option to be installed.
In time-resolved fluorometry a flash lamp is used to illuminate the sample, different
lanthanides are used as labels. The picture shows the parameters that can be set.
Protocol name
This is the name by which the protocol is identified. A password can be associated with it to
prevent parameters being changed by an unauthorized person.
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Tools menu
Flash Energy Area
There are two options for this parameter “Low” and “High”. The energy of the flash lamp
used is dependent on the discharge capacitor value. The “Low” Flash Energy Area selects a
basic capacitor of 10 nF. The “High” Flash Energy Area selects a second 10 nF capacitor in
the lamp’s discharge circuit thus doubling the energy. If you select "High" you will reduce
the counting time (number of flashes), but the counts will remain about the same.
Flash Energy Level
The Flash Energy Level sets the voltage of the discharge capacitor; the range is from 1 to
255 (corresponding to 500 to 800 volts). Note that although the lamp input energy depends
on the squared power of the discharge voltage, the output optical power is not linearly
dependent on the input energy. By changing this parameter you will fine-tune the Flash
Energy. If you increase the number, you will reduce counting time, but counts from your
sample will remain about the same.
CAUTION: If software version 2.0 release 8 or higher is installed on an instrument
having a serial number less than 4201396, then you should avoid the combination of
settings: Flash Energy Area = "High" and Flash Energy Level = high values (e.g.255)
along with 384-well or higher plates. The reason is that this "high power/continuous
usage" combination may damage the older type of flash lamp. The cooled flash excitation
system available for newer instruments is especially designed to cope with this.
Excitation Filter
The non-changeable filter in the flash excitation path., D340, has a maximum transmittance
at 340 nm and half-bandwidth of about 35 nm. The other two filters are only used for UV
absorbance.
Light Integration Capacitors
A reference circuit monitors the total excitation energy. When the selected total amount of
energy has been reached the flashes stop. The reference circuit makes use of three light
integration capacitors, numbered 1, 2 and 3, any one of which can be selected. The
capacitor selected determines the total excitation energy in the same proportions as the
capacitor numbers (within component tolerances). This means that selecting capacitor 2
instead of capacitor 1, approximately doubles the excitation energy for one measurement,
i.e. the number of flashes for one measurement doubles, hence the time used for one
measurement doubles and the number of counts doubles. Selecting capacitor 3 instead of
capacitor 1 triples the excitation energy, the measurement time and the number of counts.
The number of flashes can be seen from the result viewer by selecting
View all result data.
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Note
Each flash contributes to the total excitation energy. Flashes keep on occurring until the
required total excitation energy is achieved. However, because the energy comes in discrete
flashes the last flash will cause the total excitation energy to exceed the required amount by
a maximum of not more than the energy in one flash. This will not be significant if there are
many flashes but could be significant if there are only a few flashes. E.g. in standard
measurements the amount of flashes is about 1000, the variation in the total excitation
energy is thus 1/1000 or 0.1 %. If the amount of flashes is about 100, the variation in
excitation energies is 1/100 or 1 %.
Light Integration Reference Level
These values change the excitation energy approximately linearly. Changing this value from
50 to 100 has about the same effect as changing the capacitor from 1 to 2. The number of
flashes can be seen from the result viewer by selecting
Emission filters
View all result data.
You can select the emission filter from the drop-down list of available filters. See the
chapter on Filters for more information about how to define filters.
Factory pre-set emission filter names begin with a letter indicating the technology for which
they are to be used: D is for time-resolved fluorometry. After the letter comes the
wavelength.
Emission aperture
In front of the photomultiplier tube there is a 4-position aperture slide, which has three
different apertures and a shutter position.
The normal aperture is circular with a diameter of about 4 mm and it is used for factory set
labels.
The small aperture is circular but only 1 mm in diameter. This allows you to shift the
dynamic range of the emission signal for a particular user selected label which may
otherwise exceed the linear range of the instrument. This aperture is used for the top
fluorometry factory-set labels.
There is also a large circular aperture 5 mm in diameter.
The shutter is used at all times when no measurement is occurring to prevent stray light
getting to the photomultiplier tube.
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Counting parameters
These determine the time factors involved in the measurement process, all times are in
microseconds:
Counting delay - the time after the excitation pulse at which counting of the emission
signal begins
Counting window - the time period within which counting occurs
Counting cycle - the time between excitation pulses within a measurement.
The second or additional counting window is fully independent of the first one and it useful
when optimizing time-resolved parameters or for calculations of the decay time of the label.
Factory-set values for the main counting window
Label Delay Window Cycle
Eu 400 400 1000
Sm 50 100 1000
Tb 500 1400 2000
Dy 30 30 1000
Factory set values for the second counting window are always 0/0. The counts in this
window are displayed only if the values deviate from zero
Beam size
This parameter can be selected to be Normal or Narrow provided the Adjustable beam size
option has been installed. It enables measurements on plates with very small well sizes by
focusing the excitation light beam to a smaller size.
Second measurement parameters
In the case where you want to make measurements with two filters, you can use this
parameter to select the type of the second emission filter. The options are the same as for the
first measurement emission filter. Two measurements are made, first with the normal filter
and then with the second, which the system automatically moves into place.
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Fluorometry
Fluorometry - light of a particular wavelength is selected with a filter from the spectrum of a
continuous wave lamp and used to excite the fluorochrome in the sample. The parameters
that can be set for fluorometry are the following:
Protocol name
This is the name by which the protocol is identified. A password can be associated with it to
prevent parameters being changed by an unauthorized person.
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CW-Lamp energy
Using this parameter you are able to change the excitation energy supplied by the CWLamp.
CW-Lamp Control Mode
There are two ways of ensuring that the light incident on the plate from the CW-Lamp
remains constant – stabilized energy mode or constant voltage mode. In the stabilized
energy mode a photodiode close to the light path has part of the incident light directed onto
it. This sends feedback to the lamp controller which alters the lamp voltage to adjust the
intensity of the light produced so that it remains constant. In the constant voltage mode the
lamp voltage is not changed but the software corrects for changes in the lamp output. You
can select the control mode you want from the option buttons. The default mode is
stabilized energy. The use of the constant voltage mode allows the CW-Lamp lamp to be
run continuously at maximum power and it is faster than the stabilized energy mode. This
enables optimum performance in cases where the light intensity is critical, as in
fluorescence polarization or where speed is critical as in fast kinetics.
CW-Lamp filters
You can select the CW-lamp excitation filter from the drop-down list of available filters.
See the chapter on Filters for more information about how to define filters.
Factory pre-set CW-lamp filter names begin with a letter indicating the technology for
which they are to be used:
F is for fluorometry. After the letter comes the wavelength.
Excitation aperture (Wallac 1420-040/041/042/043 only)
There are three sizes of excitation aperture controlling the size of the incident light beam.
You can select with the option buttons Small, Normal or Large. Normal is the default.
Emission filters
You can select the emission filter from the drop-down list of available filters. See the
chapter on Filters for more information about how to define filters.
Factory preset emission filter names begin with a letter indicating the technology for which
they are to be used:
F is for fluorometry. After the letter comes the wavelength.
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Emission aperture
In front of the photomultiplier tube there is a 4-position aperture slide, which has three
different apertures and a shutter position.
The normal aperture is circular with a diameter of about 4 mm and it is used for factory set
labels.
The small aperture is circular but only 1 mm in diameter. This allows you to shift the
dynamic range of the emission signal for a particular user selected label which may
otherwise exceed the linear range of the instrument. This aperture is used for the top
fluorometry factory-set labels.
There is also a large circular aperture 5 mm in diameter.
The shutter is used at all times when no measurement is occurring to prevent stray light
getting to the photomultiplier tube.
Counter position
The counter position can be selected to be Top or Bottom.
Counting time
This parameter determines how long the excitation continues.
Second measurement parameters
In the case where you want to make measurements with two filters and/or two different
energies, you can set these parameters in this field. Click the check box
measurement
to activate them.
Second
There are three of them: CW-Lamp energy , CW-Lamp Filter and Emission Filter. See the
corresponding sections for the first measurement for more details. When you select one of
more of these parameters two measurements are made, the first with the normal settings and
the second with these new settings. The change is automatic.
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Photometry
A reference measurement is made before the plate is moved to the measurement position.
This enables the absorbance to be calculated.
The parameters that can be set for photometry are the following:
Protocol name
This is the name by which the protocol is identified. A password can be associated with it to
prevent parameters being changed by an unauthorized person.
Absorbance mode
Select whether you want visible light or ultraviolet absorbance.
Flash Lamp Filter
These option buttons are only enabled if UV absorbance mode is selected. Select the
wavelength of the filter you want. The filter will ensure that only UV light of the selected
wavelength is used for the absorbance measurement. The UV photometry filters P280 and
P260 have their transmittance peaks at 280 nm and 260 nm respectively.
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CW-Lamp filters
You can select the CW-lamp filter from the drop-down list of available filters. See the
chapter on Filters for more information about how to define filters.
Factory pre-set CW-lamp filter names begin with a letter indicating the technology for
which they are to be used:
P is for photometry. After the letter comes the wavelength.
Excitation aperture (Wallac 1420-040/041 only)
There are three sizes of excitation aperture controlling the size of the incident light beam.
You can select with the option buttons Small, Normal or Large. Normal is the default.
Reading time
This parameter determines how long the sample is read to determine the absorbance.
Second measurement
In the case where you want to make measurements with two filters, you can click this check
box and then set the parameter
CW-Lamp filter - It has the same options as the parameter
with the same name above. When you select this, two measurements are made, the first with
the normal settings and the second with this new setting. The change is automatic.
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Luminometry
Luminescence in the sample is detected.
The parameters that can be set for luminometry are show in the picture and described
below.
Protocol name
This is the name by which the protocol is identified. A password can be associated with it to
prevent parameters being changed by an unauthorized person.
Emission filters
You can select the emission filter from the drop-down list of available filters. See the
chapter on Filters for more information about how to define filters.
Factory preset emission filter names begin with a letter indicating the technology for which
they are to be used:
L is for luminescence. After the letter comes the wavelength.
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Emission aperture
In front of the photomultiplier tube there is a 4-position aperture slide, which has three
different apertures and a shutter position.
The normal aperture is circular with a diameter of about 4 mm and it is used for factory set
labels.
The small aperture is circular but only 1 mm in diameter. This allows you to shift the
dynamic range of the emission signal for a particular user selected label which may
otherwise exceed the linear range of the instrument. This aperture is used for the top
fluorometry factory-set labels.
There is also a large circular aperture 5 mm in diameter.
The shutter is used at all times when no measurement is occurring to prevent stray light
getting to the photomultiplier tube.
Counting time
This parameter determines how long the luminescence is counted for.
Second Measurement
In the case where you want to make measurements with two filters, you can click the
Second measurement check box and then set this emission filter. It has the same options as
the normal emission filter. When you select this, two measurements are made, the first with
the normal settings and the second with this new setting. The change is automatic.
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Fluorescence polarization (Wallac 1420-040/041/042/043 only)
Fluorescence polarization - light of a particular wavelength is selected with a filter from the
spectrum of a continuous wave lamp. This light is then polarized and used to excite the
fluorochrome in the sample. The emission light is then viewed through two polarizers, one
parallel to the incident polarization (S-plane) and one perpendicular to it (P-plane). These
are, each in turn, moved automatically into the emission light path. The ratio:
1000*(Parallel –G*Perpendicular) / (Parallel +G*Perpendicular)
is calculated to get the polarization in units of mP. G is a correction factor, see the end of
this section. The parameters that can be set for fluorescence polarization are the following:
Protocol name
This is the name by which the protocol is identified. A password can be associated with it to
prevent parameters being changed by an unauthorized person.
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CW-Lamp energy
In fluorescence polarization constant voltage mode is used and the light source is driven
with high power. The software corrects for changes in energy of the CW-lamp to ensure that
results are calculated for a constant excitation energy.
should always use maximum power, 65535
CW-Lamp filter
.
For fluorescence polarization you
You can select the CW-lamp excitation filter from the drop-down list of available filters.
See the chapter on Filters for more information about how to define filters. Factory pre-set
CW-lamp filter names begin with a letter indicating the technology for which they are to be
used: F is for fluorometry. After the letter comes the wavelength.
Polarizer aperture
There are two sizes for the aperture of the S-plane polarizer in the excitation light path. You
can select by option buttons whether this is Small (for high-density plates) or Normal (96 or
384-well plates).
Emission filter
You can select the emission filter from the drop-down list of available filters. See the
chapter on Filters for more information about how to define filters. Factory preset emission
filter names begin with a letter indicating the technology for which they are to be used: F is
for fluorometry. After the letter comes the wavelength.
Emission aperture
In front of the photomultiplier tube there is a 4-position aperture slide, which has three
different apertures and a shutter position. The normal aperture, which is circular with a
diameter of about 4 mm, should be used for fluorescence polarization.
Counting time
This parameter determines how long the excitation continues. The following times are
recommended for 96 and 384-well plates:
Concentration (nM) Measuring time (s)
2 or more 0.1
1 0.2
0.5 0.3
0.1 1.0
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×
−
G factor
In fluorescence polarization assays a G-factor is used to correct for the bias in polarization
values caused by differences in instruments and assay conditions. To calculate the G-factor
prepare buffer and 1 nM fluorescein samples (replicates should be at least 24). The formula
used is:
L
CorrectedS
G
=
CorrectedP
1(
−
*
1(
+
1000
L
)
)
1000
where the corrected parallel and perpendicular S- and P-counts are background subtracted
fluorescence count rates obtained from a pure fluorophore solution e.g.:
CorrectedS = averageS(1nM)-averageS(buffer) and
CorrectedP = averageP(1nM)-averageP(buffer)
L is the literature polarization value for the fluorophore in mP. E.g. the value for 1 nM
fluorescein is 27 mP in aqueous buffer at room temperature. The G-factor is typically
between 0.8 and 1.2.
The G factor is used in the calculation of the corrected mP for samples as:
mP
where CorrectedS = S(sample)-S(buffer) and
×= 1000
CorrectedP = P(sample)-P(buffer)
CorrectedPGCorrectedS
CorrectedPGCorrectedS
×+
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LANCE
Note: this requires the LANCE option to be installed.
In LANCE homogenous time-resolved fluorescence a flash lamp is used to illuminate the
sample. In the most common form of LANCE this excites the donor molecule which, after a
delay, transfers the energy to the accepter molecule which then emits light. Two labels must
be defined, one for the donor and the other for the accepter. The picture shows the
parameters that can be set for a LANCE label.
Protocol name
This is the name by which the protocol is identified. A password can be associated with it to
prevent parameters being changed by an unauthorized person.
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Flash Energy Area
There are two options for this parameter “Low” and “High”. The energy of the flash lamp
used is dependent on the discharge capacitor value. The “Low” Flash Energy Area selects a
basic capacitor of 10 nF. The “High” Flash Energy Area selects a second 10 nF capacitor in
the lamp’s discharge circuit thus doubling the energy. If you select "High" you will reduce
the counting time (number of flashes), but the counts will remain about the same.
Flash Energy Level
The Flash Energy Level sets the voltage of the discharge capacitor; the range is from 1 to
255 (corresponding to 500 to 800 volts). Note that although the lamp input energy depends
on the squared power of the discharge voltage, the output optical power is not linearly
dependent on the input energy. By changing this parameter you will fine-tune the Flash
Energy. If you increase the number, you will reduce counting time, but counts from your
sample will remain about the same.
CAUTION: If software version 2.0 release 8 or higher is installed on an instrument
having a serial number less than 4201396, then you should avoid the combination of
settings: Flash Energy Area = "High" and Flash Energy Level = high values (e.g.255)
along with 384-well or higher plates. The reason is that this "high power/continuous
usage" combination may damage the older type of flash lamp. The cooled flash excitation
system available for newer instruments is especially designed to cope with this.
Excitation filter
The non-changeable filter in the flash excitation path., D340, has a maximum transmittance
at 340 nm and half-bandwidth of about 35 nm. The other two filters are only used for UV
absorbance.
Light integration capacitors
A reference circuit monitors the total excitation energy. When the selected total amount of
energy has been reached the flashes stop. The reference circuit makes use of three light
integration capacitors, numbered 1, 2 and 3, any one of which can be selected. The
capacitor selected determines the total excitation energy in the same proportions as the
capacitor numbers (within component tolerances). This means that selecting capacitor 2
instead of capacitor 1, approximately doubles the excitation energy for one measurement,
i.e. the number of flashes for one measurement doubles, hence the time used for one
measurement doubles and the number of counts doubles. Selecting capacitor 3 instead of
capacitor 1 triples the excitation energy, the measurement time and the number of counts.
The number of flashes can be seen from the result viewer by selecting
View all result data.
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Note
Each flash contributes to the total excitation energy. Flashes keep on occurring until the
required total excitation energy is achieved. However, because the energy comes in discrete
flashes the last flash will cause the total excitation energy to exceed the required amount by
a maximum of not more than the energy in one flash. This will not be significant if there are
many flashes but could be significant if there are only a few flashes. E.g. in standard
measurements the amount of flashes is about 1000, the variation in the total excitation
energy is thus 1/1000 or 0.1 %. If the amount of flashes is about 100, the variation in
excitation energies is 1/100 or 1 %.
Light Integration Reference Level
These values change the excitation energy approximately linearly. Changing this value from
50 to 100 has about the same effect as changing the capacitor from 1 to 2. The number of
flashes can be seen from the result viewer by selecting
Emission filter
View all result data.
You can select the emission filter from the drop-down list of available filters. See the
chapter on Filters for more information about how to define filters.
Factory pre-set emission filter names begin with a letter indicating the technology for which
they are to be used: D is for time-resolved fluorometry. After the letter comes the
wavelength.
Emission aperture
In front of the photomultiplier tube there is a 4-position aperture slide, which has three
different apertures and a shutter position.
The normal aperture is circular with a diameter of about 4 mm and it is used for factory set
labels.
The small aperture is circular but only 1 mm in diameter. This allows you to shift the
dynamic range of the emission signal for a particular user selected label which may
otherwise exceed the linear range of the instrument. This aperture is used for the top
fluorometry factory-set labels.
There is also a large circular aperture 5 mm in diameter.
The shutter is used at all times when no measurement is occurring to prevent stray light
getting to the photomultiplier tube.
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Counting parameters
These determine the time factors involved in the measurement process, all times are in
microseconds:
Counting delay - the time after the excitation pulse at which counting of the emission
signal begins
Counting window - the time period within which counting occurs
Counting cycle - the time between excitation pulses within a measurement.
The second or additional counting window is fully independent of the first one and it useful
when optimizing time-resolved parameters or for calculations of the decay time of the label.
Factory-set values for the main counting window
Label Delay 1 Delay 2 Window 1 Window 2 Cycle
High count 615 50 0 100 0 1000
High count 665 50 0 100 0 1000
Long decay 615 50 0 100 0 3000
Long decay 665 50 0 100 0 3000
545 50 0 100 0 3000
615 50 0 100 0 1000
665 50 0 100 0 1000
572 50 0 100 0 3000
TruPoint 615 50 200 100 100 1000
TruPoint 545 50 200 100 100 1000
Beam size
This parameter can be selected to be Normal or Narrow provided the Adjustable beam size
option has been installed. It enables measurements on plates with very small well sizes by
focusing the excitation light beam to a smaller size.
Second measurement parameters
In the case where you want to make measurements with two filters, you can use this
parameter to select the type of the second emission filter. The options are the same as for the
first measurement emission filter. Two measurements are made, first with the normal filter
and then with the second, which the system automatically moves into place.
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LANCE Label Properties
• LANCE 615: 100 flashes, window 50/100/1000, 615nm. This label is for fast decay
Eu-chelates (for example Eu-W1024) in a TR-FRET assays.
•
• LANCE 665: 100 flashes, window 50/100/1000, 665nm. This label is for fast decay
Eu-chelates (for example Eu-W1024) in a TR-FRET assays.
•
• LANCE High Count 615: (same as LANCE 615 but 1000 flashes). Higher count rate.
•
• LANCE High Count 665: (same as LANCE 665 but 1000 flashes). Higher count rate.
•
• LANCE Long Decay 615: (same as LANCE 615 but 3000 cycle time). This label is
for long decay Eu-chelates (for example Eu-W8044) in a TR-FRET assays.
•
• LANCE Long Decay 665: (same as LANCE 665 but 3000 cycle time). This label is
for long decay Eu-chelates (for example Eu-W8044) in a TR-FRET assays.
•
• LANCE 545 (same as LANCE Long Decay 615 but 545nm emission).
•
• LANCE 572(same as LANCE Long Decay 665 but 572nm emission).
•
• TruPoint 615 (same as LANCE 615 but including 2
nd
window 200/100) This is a label
for Eu-TRF-Quenching assays.
•
• TruPoint 545 (same as LANCE 615 but with 545 emission filter and 2
nd
window
200/100) This is a label for Tb-TRF-Quenching assays.
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Filters
In the enhanced security mode this tool is enabled only for users who belong to the Editor
user group.
In this window there are three tabs: "Emission Filters", "CW-Lamp Filters" (i.e. continuous
wave lamp excitation filters) and "Filter Slides" (including filter wheels). "Filter Slides"
allows you to specify the slides for emission and CW-lamp filters. The names of the filters
or slides in the currently selected tab are displayed. Names of user defined filters can be
freely defined by the user but names of factory preset filters cannot be changed.
There are four buttons at the bottom of the window:
Add - add a new item to the list available e.g. a new filter (not available for slides). Give the
name and edit the properties. It will appear with a new icon showing it is different from a
default item.
Copy - (select an item to activate this, not available for slides). Make a copy of an item. It
will appear with a new icon showing it is different from a default item, and the name will be
preceded by the words “Copy of”. You can give it a different name and edit other
parameters by selecting
Remove - (select a user created item to activate this; not available for slides). Remove the
selected item from the list. You cannot remove a pre-set item from the list nor can you
remove a filter that has been assigned a position in a slide. This command requires
confirmation because you cannot undo
Properties.
Remove.
Properties - (select an item to activate this). View the properties of the selected item. If the
item is user defined then you can edit the properties, but if it is a factory pre-set, you cannot
edit them.
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Emission filters
If you select properties of an emission filter or you want to define the properties of a new
filter, a further dialogue appears where the name of the filter and the technology for its uses
can be defined. There is also a section where you can add additional information that you
can connect with the filter properties.
Factory pre-set emission filter names begin with a letter indicating the technology for which
they are to be used:
D is for time-resolved fluorometry (LANCE filters are marked as such).
F is for fluorometry.
L is for luminescence.
After the letter comes the wavelength.
.
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CW-Lamp filters
If you select properties of a CW-lamp filter or you want to define the properties of a new
excitation filter, a further dialogue appears where the name of the filter and the technology
for its use can be defined. There is also a section where you can add additional information
that you can connect with the filter properties.
Factory pre-set CW-lamp filter names begin with a letter indicating the technology for
which they are to be use:
F is for fluorometry
P is for photometry (absorption)
After the letter comes the wavelength.
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Filter slides
There are two types of slide: Emission filter and CW-lamp excitation filter. The former is a
strip and the latter a wheel. The instrument is loaded with slide A in each case but an empty
slide B is also available for user filters as described below.
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Emission filter slide
There are 8 positions in emission filter slides A and B. The physical dimensions of the filter
slots are:
diameter 25.5 mm
depth 11 mm
clear aperture 22 mm.
All standard filters with diameter of 25.4 mm (1 inch) and thickness of 10 mm fit in the
emission filter slides.
In emission filter slide A, positions 1 to 4 and 8 are used for emission filters for timeresolved fluorescent chelates (if this option is included), position 5 and 6 for emission filters
for fluorometry labels umbelliferone and fluorescein, respectively. Position 7 is kept open
for luminometry measurement. If you want to add you own filters you must either use slide
B which is fully customizable, or change some of the existing filters.
Factory-pre-set filter names begin with a letter indicating the technology.
D is for Time resolved fluorometry, including LANCE
F is for fluorometry
P is for photometry (not used for emission filters)
L is for luminometry
After the letter comes the wavelength:
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User-defined filters can be named freely but there is only space for five letters to be shown
in the figure of the filter position. The full name will be shown if you move the mouse
cursor onto the slot where the filter is.
Physically changing the emission filter slide
Click the Eject slide button on the Emission filter slide dialogue. A message will appear
telling you to unscrew and remove the plate on the side of the counter. When you have
removed the plate, click
Avoid touching the filters with your fingers.
OK to cause the slide to be ejected. Pull the slide out carefully.
When you have got the slide out you can then change the filters or load filters into emission
filter slide B.
When the slide is ready, carefully return the slide to its position. Push it in as far as it will
go. Replace the filter slide cover and screw it in place, then click
OK on the dialogue on the
screen. After this you can operate the instrument.
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CW-Lamp filter wheel
Excitation filters fit into a wheel positioned in front of the CW lamp. There are eight
positions on this wheel and two are reserved by default for fluorometry and three for
photometry. The remaining positions can have filters added by the user.
Note: default filters can be removed and replaced by user filters.
There is also a second filter wheel (B) which has only four positions but these are for the
most common size of commercial filters i.e. “round one inch” and the wheel is supplied
empty with locking rings.
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Physically changing the CW-Lamp filter wheel
If you want to change a filter in the CW-Lamp filter wheel or you want to install wheel B
then you must open the cover and lift out the filter wheel as shown in the picture. Either
change the filters you want or take a new filter wheel B. Replace the filter wheel and make
sure you close the cover before starting a measurement.
Changing the CW-Lamp
If the CW-Lamp needs replacing - lift the cover to get access to the lamp. Make sure the
lamp is switched off and give the lamp chance to cool
before removing it and replacing it.
Follow the instructions on the inside of the cover, see the picture on the next page. You will
probably find it easier to use your left hand to remove the lamp from the clips holding it.
When you have replaced the lamp, close the cover and switch power on.
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Changing filters in the software
Note: if you change a filter in the software you must also change it physically in the filter
slide or wheel.
When you select Properties, a picture of the selected slide appears with the filters that are
loaded in it marked in red or black as shown previously. Those that are in red are factory
pre-set. The excitation filter slide is in the form of a wheel and the emission filter slide is a
strip.
To move a filter from the list to a place on the slide, move the cursor to the filter name on
the list, drag it keeping the mouse button pressed down until it is over the empty slide
position that you want it to occupy, then release the mouse button. The name of the filter
will appear in black in the slide position.
To move a filter from one position to another, click on it and drag it to the empty place
you want it to occupy.
To remove a filter from the slide, click on it and drag it back to anywhere in the area of
the
Available filters list, then release the mouse button.
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