Eppendorf AF2200 User Manual
ualPlateReader AF2200
E
Register your instrument!
www.eppendorf.com/myeppendorf
N)manual
Eppendorf® PlateReader AF2200
Software manual
Version 1.0.4
©
Copyright
2013 Eppendorf AG, Hamburg. No part of this publication may be reproduced without the prior
permission of the copyright owner.
Trademarks
Eppendorf
Microsoft
®
and the Eppendorf Logo are registered trademarks of Eppendorf AG, Hamburg, Germany.
®
, Windows® and Excel® are registered trademarks of Microsoft Corporation, Redmond, WA,
USA.
Pentium
PicoGreen
®
is a registered trademark of Intel Corporation, Santa Clara, CA, USA.
®
, RiboGreen®, OliGreen® and NanoOrange® are registered trademarks of Molecular Probes,
Inc., Eugene, OR, USA.
®
Apo-ONE
Corning
Greiner
und CellTiter-Blue® are registered trademarks of Promega Corporation, Madison, WI, USA.
®
is a registered trademark of Corning Incorporated, New York, NY, USA.
®
and μClear® are registered trademarks of Greiner Labortechnik GmbH, Frickenhausen, Germany.
Lumox™ is a trademark of Greiner Bio-One GmbH, Frickenhausen, Germany.
Nunclon
®
is a registered trademark of Nunc A/S, Roskilde, Denmark.
LumiNunc™ und FluoroNunc™ are trademarks of, Nunc A/S, Roskilde, Denmark.
Trademarks are not marked in all cases with ™ or
6141 905.200-01/022013
®
in this manual.
Table of contents
®
Eppendorf
PlateReader AF2200
English (EN)
Table of contents
1 Operating instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.1 Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2 Danger symbols and danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2.1 Danger symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2.2 Danger levels. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.3 Symbols used . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
2 Product description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
2.1 Area of Application . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
2.2 System requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
2.3 CE Declaration for Europe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3 Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.1 Install software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.2 Starting PlateReader AF 2200. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
3.2.1 Connected Instrument. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
3.2.2 Simulated Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
3
4 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
4.1 Main Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
4.2 Explorer bar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
4.3 Workflow Pane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
4.4 Info bar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
4.5 Menu Bar. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
4.5.1 File Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
4.5.2 Edit Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
4.5.3 View Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
4.5.4 Instrument Menu. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
4.5.5 Settings Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
4.6 Plate List . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
5 Defining measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
5.1 Plate Size – Pattern . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
5.2 Defining Endpoint Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
5.3 Defining Multimode Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
5.4 Defining Kinetic Measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
5.5 Indenting and Releasing Program Elements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Table of contents
®
4
Eppendorf
PlateReader AF2200
English (EN)
6 Predefined methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
6.1 Method selection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
6.2 Method description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
6.2.1 Nucleic acid quantification (UV 260 nm microvolume) . . . . . . . . . . . . . . . . . . . . . . . . 54
6.2.2 Nucleic acid quantification (UV 260 nm with factor) . . . . . . . . . . . . . . . . . . . . . . . . . . 54
6.2.3 Nucleic acid quantification (UV 260 nm with standards) . . . . . . . . . . . . . . . . . . . . . . . 56
6.2.4 Nucleic acid quantification (Fluorescence 485/535 nm with standards) . . . . . . . . . . . 59
6.2.5 Protein quantification BCA, Bradford, Lowry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
6.2.6 Protein quantification (NanoOrange 485/595 nm with standards) . . . . . . . . . . . . . . . . 63
6.2.7 Cell viability (CellTiter-Blue Assay). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
6.2.8 Cell viability (Apo-ONE Caspase-3/7 Assay) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
6.3 Method (measurement) parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
6.4 Method results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
6.4.1 Contents of the raw data sheet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
6.4.2 Contents of the blanking sheet (factor methods only) . . . . . . . . . . . . . . . . . . . . . . . . . 70
6.4.3 Content of the results sheet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
7 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
7.1 General Troubleshooting. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
7.2 Troubleshooting Absorbance mode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
7.3 Troubleshooting Fluorescence mode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
7.4 Save method scripts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
7.5 Save method results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
8 Evaluation procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
8.1 Nucleic acid quantification UV 260 nm with factor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
8.2 Nucleic acid and Protein quantification with standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
8.3 Protein quantification Nano Orange 485/595 nm with standards . . . . . . . . . . . . . . . . . . . . . . . 75
8.4 Cell viability CellTiter-Blue Assay, Cell viability Apo-ONE Caspase-3/7 Assay . . . . . . . . . . . . . 75
Operating instructions
®
Eppendorf
PlateReader AF2200
English (EN)
1 Operating instructions
1.1 Using this manual
Read this operating manual completely before using the device for the first time. Please also note the
operating instructions for the accessories, if applicable.
This operating manual is part of the product. Thus, it must always be easily accessible.
Enclose this operating manual when transferring the device to third parties.
If this manual is lost, please request another one. For the current version, please refer to our webpage
www.eppendorf.com/worldwide
(international) or www.eppendorfna.com (North America).
1.2 Danger symbols and danger levels
The safety instructions in this manual appear with the following danger symbols and danger levels:
5
1.2.1 Danger symbols
Biohazard Explosion
Electric shock Toxic substances
Hazard point Material damage
1.2.2 Danger levels
DANGER Will lead to severe injuries or death.
WARN ING May lead to severe injuries or death.
CAUTION May lead to light to moderate injuries.
NOTICE May lead to material damage.
Operating instructions
Eppendorf
6
English (EN)
®
PlateReader AF2200
1.3 Symbols used
Symbol Meaning
Handling
1.
2.
• List
Text Name of fields in the software
Actions in the specified order
Useful information
Product description
®
Eppendorf
PlateReader AF2200
English (EN)
2 Product description
2.1 Area of Application
The Software is an easy-to-use and flexible tool, which gives the user complete control over the Eppendorf
PlateReader AF2200.
The Software presents the raw data for further use in Excel, offering excellent features for research
purposes.
2.2 System requirements
To install the software, the system must meet the following minimum requirements:
Component Minimum Recommended
7
Computer Windows XP (32-bit):
Windows-compatible computer
with a Pentium-compatible
processor, 1 GHz
Windows Vista (32-bit):
Windows-compatible computer
with a Pentium-compatible
processor, 1 GHz
Windows 7 (32- or 64-bit):
Windows-compatible computer
with a Pentium-compatible
processor, 1 GHz
Operating system Windows XP (32-bit) SP3
Windows Vista (32-bit)
Windows 7 (32-bit)
Windows 7 (64-bit)
Memory Windows XP: 512 MB RAM
Windows Vista (32-bit): 1 GB
RAM
Windows 7 (32-bit): 1 GB RAM
Windows 7 (64-bit): 2 GB RAM
2 GHz (dual-core)
2 GHz (dual-core)
2 GHz (dual-core)
Windows XP (32-bit) SP3
1 GB RAM
2 GB RAM
2 GB RAM
3 GB RAM
Memory requirements 700 MB RAM 1 GB RAM
Monitor Super VGA Graphics 1280 x 1024
Screen resolution 1024 x 768 -
Color depth 256 -
Mouse Microsoft-compatible mouse or
similar device
-
Product description
Eppendorf
8
English (EN)
Component Minimum Recommended
Communication 1x USB 2.0 2x USB 2.0
®
PlateReader AF2200
1x RS232 (serial)
Drive, graphic card 1xCD-ROM drive
Windows Vista:
DirectX 9 graphic and 32 MB
graphics memory (for Home Basic
version)
DirectX 9 graphic and 128 MB
graphics memory with WDDM
support for all other versions.
Windows 7:
DirectX 9 graphic with WDDM
driver 1.0 or higher.
.NET Microsoft .NET Framework 2.0
If this version is not available, it
will be installed by the installation
and update program using any
available installation of the .NET
Framework.
Windows Installer 3.1
If this version is not available, it
will be installed by the installation
and update program.
Microsoft Excel Excel 2002 (only compatible for
script-based messages without
data reduction). Excel 2002 is not
compatible for methods.
Excel 2003, 2007 and 2010
(32-bit)
The Starter Edition is not
supported.
-
-
-
-
2.3 CE Declaration for Europe
PlateReader AF2200 Software is not a CE-marked product. Therefore no CE declaration for Europe is
available.
3 Installation
3.1 Install software
You must have administrator rights to install the software.
Install the software before connecting the PlateReader AF2200 to the computer.
®
Eppendorf
1. Insert the CD with the software into the CD-ROM
drive.
2. Start the installation by executing the Setup file.
PlateReader AF2200
Installation
English (EN)
9
3. Press the OK button to continue the installation.
4. Press the Next button to continue the installation.
The Next button can be used to confirm the current installation step and continue the
installation.
The Back button brings you back to the previous installation step.
The Cancel button can be used to abort the installation.
10
Installation
Eppendorf
®
English (EN)
PlateReader AF2200
5. Define the user.
6. Select language.
7. Define the installation directory.
®
Eppendorf
PlateReader AF2200
English (EN)
8. Press the Install button to confirm the settings
and start the installation.
9. Press the Finish button to complete the
installation.
Installation
11
The PlateReader AF2200 can now be connected to the computer.
12
Installation
Eppendorf
English (EN)
®
PlateReader AF2200
3.2 Starting PlateReader AF 2200
The Software can be used either with a connected instrument or in simulation mode.
3.2.1 Connected Instrument
Install the software before connecting the instrument to the computer.
1. Connect the instrument to your computer and switch the instrument on.
2. Start the program by selecting Programs > Eppendorf > PlateReader AF2200 from the Windows Start
menu.
3. The following dialog box appears:
4. Select the instrument name in the dialog box.
5. Click the Button OK to start the software.
3.2.2 Simulated Instrument
1. Start the program by selecting Programs > Eppendorf > PlateReader AF2200 from the Windows Start
menu.
2. The following dialog box appears:
3. Select the checkbox Show simulated instruments.
4. Select the demo instrument to connect to from the Instrument Name list.
5. Click the Button OK to start the software.
Overview
1
2
3
456
English (EN)
Eppendorf
®
PlateReader AF2200
4Overview
4.1 Main Window
The main window of the software is used to set up workflows. Each workflow is easily created by dragging
and dropping the process steps into a sequence according to the application. The application workflow is
then visible to the user in the workflow pane and can be saved for future use.
Each process step, that is each program element, can be copied and pasted and moved to the desired
position in the workflow.
Data can be exported easily to Windows compatible formats (Excel).
Start the software and connect an instrument as described in the previous chapter or select the simulation
mode. The Software main window appears.
13
1Menu bar
2Workflow pane
3Info bar
4 Controls
5 Status bar
6Explorer bar
14
Overview
Eppendorf
English (EN)
®
PlateReader AF2200
4.2 Explorer bar
The Explorer bar is divided into different sections. Each section contains program elements used to create
an individual workflow.
1. Create a workflow either by double-clicking the selected program element or by dragging and dropping
it into the workflow pane.
The following program elements are available:
Methods Eppendorf prdefined methods
Labware Plate
Pattern
Well
Mode Absorbance
Fluorescence Intensity
Actions Temperature
Shaking
Move Plate
Kinetic Kinetic Cycle
Kinetic Condition
Miscellaneous Comment
User Intervention
Wait for Temperature
Wait Time
Incubation
4.2.1 Labware
Plate
The Plate program element is used to select a plate format from the Plate definition drop-down list.
For accurate measurements it is essential that plate and pattern selected in the software
correspond to the plate and the pattern you use for measuring.
1. Click Details to see further information on the selected plate.
2. If a plate with cover is used, select the Plate with cover checkbox.
3. The measurement will automatically measure all selected wells of the plate. If you want to measure a
specific well or a range of wells, click the link Pattern in the lower right corner.
®
Eppendorf
PlateReader AF2200
English (EN)
4. Under Details it is possible to apply a filter so that only certain plate definition files are shown.
Pattern
The Pattern program element is displayed according to the selected plate format (number of wells).
1. To measure individual wells, click the desired well or to measure a range of wells drag a frame around
the desired range.
Overview
15
Independent Pattern
1. Clicking on Zoom, the plate preview can be zoomed and independent parts of the plate can be selected
2. A second range of wells can be selected by pressing the Control key on the keyboard and dragging a
frame over the wells to be selected.
16
Overview
®
Eppendorf
PlateReader AF2200
English (EN)
Well
Use the Well program element to perform measurements well by well. Without this program element, all
measurement steps are done plate-wise.
4.2.2 Mode
Absorbance
The Absorbance program element is used to perform absorbance measurements.
1. Enter or select the respective parameters:
Wavelength Specify a measurement wavelength.
The Reference wavelength may be selected to correct for flash
variations.
Two drop-down lists display the available measurement and
reference filter wavelengths, according to the inserted absorbance
filter slide. If the drop-down lists are empty, the absorbance filter
either has not been inserted into the reader or has not been defined.
Read Specify a certain Number of flashes and, if required, a Settle time
before the next measurement. The number of flashes is selectable
from 1 – 100.
Settle time: Enter a value to specify the time between the movement
of the plate carrier to the measurement position and the first flash.
Flashes In order to obtain a good measurement precision it is recommended
to perform absorbance measurements with the number of flashes
that is set as a default value for the respective instrument. On-the-fly
measurements with one flash per well are possible with all plate
types.
Measurement Enter a measurement name.
®
Eppendorf
PlateReader AF2200
English (EN)
Fluorescence Intensity
The Fluorescence Intensity program element contains fields for the selection of excitation and emission
wavelength, top or bottom reading mode, integration and lag time, flash number and gain settings. A
checkbox for multiple reads per well gives access to additional function.
Overview
17
The following are the Fluorescence Intensity parameters:
Wavelength and Bandwidth Specify an Excitation and an Emission wavelength.
Two drop-down lists display the available measurement filter
wavelengths.
If the spin box of fixed values is empty, the excitation and emission
filters have not been inserted into the reader or have not been
defined.
The bandwidth is specific for every used filter and needs to be
defined together with the central filter wavelength.
Read Specify a certain Number of flashes and, if required, a Settle time
before the next measurement. The number of flashes is selectable
from 1 – 100.
Settle time: Enter a value to specify the time between the movement
of the plate carrier to the measurement position and the first flash.
Flashes In order to obtain a good measurement precision it is recommended
to perform fluorescence measurements with the number of flashes
that is set as a default value for the respective instrument. On-the-fly
measurements with one flash per well are possible with all plate
types.
Mode Select Top or Bottom.
Measurement Enter a measurement name
18
Overview
®
Eppendorf
PlateReader AF2200
English (EN)
Gain The gain is an amplification factor for the photomultiplier tube (PMT)
and may be set by selecting one of the following modes:
Manual gain: user-defined gain value (valid range: 1-255)
Optimal gain: calculated automatically by the instrument according
to the highest signal within the selected well range in order to avoid
OVER. Optimal gain determination is performed in a
pre-measurement. It is recommended to use the optimal gain
function for all applications that produce results with unknown RFU
values.
Calculated from well: determines the optimal gain for the selected
well. The resulting gain value is applied to all other wells within the
selected well range.
Extended dynamic range: The extended dynamic range option is an
automatic gain function that serves to optimally adjust the gain
setting for both very high and very low signals on a microplate within
one single measurement. By selecting "extended dynamic range",
the measurement is done in two consecutive parts, one with a high
and one with a low gain. The results of both measurements are
automatically correlated and displayed within one single data set.
For measurements in combination with the Extended dynamic range function, a very high gain
value in the first measurement can result in OVER values in the second measurement. When
only very high and very low signal intensities are detected within the measured well range, the
new gain value is calculated on the basis of the very low values, in certain cases this can lead
to OVER for the entire second measurement.
Integration/Lag time Integration time: duration of signal recording per well (valid range:
20-2000 μs).
Lag time: time between flash and the start of signal integration.
While lag time is an optional function, the integration time is a
mandatory parameter for defining the duration of signal recording.
The default values for standard fluorescence intensity measurements
are 0 μs lag time and 20 μs integration time.
®
Eppendorf
PlateReader AF2200
English (EN)
Multiple Reads per Well
The software allows the user to define multiple reads per well (MRW) in Absorbance, Fluorescence top and
Fluorescence bottom mode.
The MRW feature is not available for well wise measurements.
The Reference wavelength on the absorbance program element is not selectable in combination with
multiple reads per well.
The multiple reads per well function can be activated on an absorbance or fluorescence intensity program
element by selecting the Multiple reads per well check box.
Overview
19
More details on defining parameters for multiple reads per well, are available in the Operating
Manual of the Eppendorf PlateReader AF2200.
The multiple reads per well function is available for plate formats with up to 384 wells.
4.2.3 Actions
Tem perature
1. Select the Temperature program element to enter a certain target temperature.
2. Select On to enter a target temperature value.
3. Click on the link Wait until temperature is reached to define the Minimum and/or Maximum temperature
values. The heating of the instrument starts when clicking the Start button.
4. For pre-heating the instrument, select Heating in the Instrument menu and click the On button.
The measurement only starts if the current instrument temperature is within the specified range (see
Miscellaneous on p. 21).
20
Overview
®
Eppendorf
PlateReader AF2200
English (EN)
Shaking
1. Select the Shaking program element if the plate is to be shaken, either before the measurement or
between kinetic cycles.
2. Enter the respective parameters (see Parameter list).
3. Clicking the link Wait a couple of seconds inserts a new program element (see Miscellaneous on p. 21).
Tab. 4-1: Parameter list
Duration Enter the duration of the shaking process.
Mode Select between the options Linear and Orbital.
Amplitude Enter the required Amplitude value from the drop-down list.
4.2.4 Kinetic
Kinetic Cycle
Use the program element Kinetic Cycle to perform several consecutive measurements, which may be
executed in certain intervals.
1. Enter the respective parameters:
Cycles Number of cycles: Enter a number or click the up or down arrows for
the number of actual measurement steps (2 – 1000 cycles)
Duration: Enter the duration, format hh:mm:ss.
Kinetic Interval Use kinetic interval: Enter the time interval (hh:mm:ss or ms).
Plate-wise kinetic measurements
Each cycle of the kinetic measurement is performed on all selected wells. Plate-wise kinetic measurements
may contain a maximum of ten independent measurement stripes that do not need to be of the same
measurement type.
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Well-wise kinetic measurements
All cycles of the kinetic measurement are first performed in one well before continuing to the next well.
Well-wise kinetic measurements may be composed of a maximum of four measurement stripes of the same
type, e.g., four absorbance stripes.
Kinetic Condition
Use the Kinetic Condition program element to define which actions should be executed at a certain cycle.
If 3 is entered for Execute command at cycle within a kinetic measurement containing, e.g. a Shake step,
shaking is performed only at cycle 3.
Overview
21
Kinetic conditions should be inserted right after a Kinetic Cycle program element in order to
ensure optimal result reproducibility.
Users are advised to set up suitable scripts prior to the measurements and to use the same
script for all similar kinetic measurements in order to obtain comparable results.
4.2.5 Miscellaneous
Comment
Use the program element Comment to enter a remark or statement for the current measurement in the text
field. This text is shown together with the measurement in the Excel output sheet.
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Overview
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User Intervention
The User Intervention program element informs the operator of the instrument to execute a definite action
during the workflow at a certain time.
If for example the Move Plate program element is used to move the plate out to perform a certain action,
then the entered text should inform the operator to perform these actions. A dialog box shows the message
and the measurement process stops until OK is clicked.
If the plate should be moved in after pipetting for example, then the text Move Plate In informs the operator
to move the plate in after pipetting to continue the workflow.
Wait for Temperature
Use the program element Wait for Temperature to define a valid temperature range for the assay.
This is typically used after a Temperature program element
Wait Time
Use the Wait Time program element to define a certain waiting period before the next step within a
workflow is executed.
In the Wait time field enter the required time.
1. Enter the respective parameters:
Timer Enter the Wait time (hh:mm:ss)
Options Ignore wait at last kinetic cycle: When the program step Wait Time is
the last action within a kinetic run, the wait time will be ignored in
the last cycle.
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Incubation
Incubation is always done at the heating position to ensure inside the instrument proper temperature
distribution.
Incubation can consist of shaking and waiting steps (up to 2 shaking steps and up to 2 waiting steps are
allowed in any combination).
The Remaining Wait step waits until the overall incubation time is over (including shaking and waiting
times).
The incubation program element is typically used to perform shaking and waiting at a certain temperature
for a certain time.
Overview
23
1. Enter the appropriate parameters for incubation:
Incubation time Enter the total time (min. 5 s)
Actions Available actions: Shaking, Wait Time
2 wait and 2 shaking actions are allowed. Select actions by
double-clicking or use the arrow keys.
Organize actions by using the up/down keys.
Remaining Wait Time: mandatory, cannot be deleted or edited
(duration 3 s)
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Overview
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PlateReader AF2200
4.3 Workflow Pane
The main window in the software is the Workflow pane, where the measurement script is visible and where
parameters are defined and edited.
There are two ways to insert a program element from the Explorer bar into the Workflow pane.
1. Select a program element from the Explorer bar; by double-clicking it, it is inserted into the Workflow
pane directly after the previous program element.
2. Click the program element in the Explorer bar and drag it into the Workflow pane to the respective
position.
The program elements are numbered according to their sequence.
Once a program element has been inserted into the Workflow pane, settings and parameters for this
element can be entered or edited.
Single program elements inside the Workflow pane can be collapsed to display the most important
information or expanded to access all editable functions.
3. Click one of the triangles next to the title of the program element, or , to switch between the two view
modes.
By default, the software starts with the Plate element in the Workflow pane. This can be modified in the
Settings menu > User Settings.
Currently selected program elements within the Workflow pane are displayed with a blue heading.
If a program element contains errors or is invalid within the current workflow, the element will
be flagged with an error mark and the number of the element is highlighted in red. In the
Status bar, the number of Errors appears in red. In the Info bar detailed information on the
error is displayed. If the workflow contains errors, the measurement script can neither be
saved nor started.
It is recommended to always save the workflow before starting a measurement. You can define this feature
as default in the Settings menu > User Settings > Options.
Select Save the script before it is started.
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