Addendum 2005-09-22 - GE Healthcare Life Sciences

Addendum 2005-09-22 - GE Healthcare Life Sciences
Addendum 2005-09-22
Scanning Membranes
1 Loading a membrane
When scanning membranes, the cassette with permanently mounted glass plate
- Mounted Glass Cassette - should be used. To achieve optimal calibration when
scanning, the black plastic hold down plate should be placed on top of the
membranes.
Note: Wear powder free gloves when handling the cassette and membranes. The
powder used in laboratory gloves can fluoresce and may also scatter light
affecting image quality.
To load a cassette
1 Clean the glass plate with distilled water and dry it using a lint free cloth.
Since fluorescent material has come into direct contact with the glass plate
we recommend using a lint free tissue moistened with 10% hydrogen
peroxide to remove this material, followed by cleaning with distilled water.
2
Place the membrane(s) directly on the plate with the blotting side facing
down. If the membrane(s) is wet make sure to exclude air bubbles when
positioning it.
3
Note the positions of the membrane boundaries on the plate. You will use this
information to determine the area(s) to be scanned, see 2 Set Up a membrane
scan.
4
Place the hold down plate in the cassette on top of the membranes.
5
Put the lid on the cassette (see Fig. 3-4 in the User Manual), and close the
cassette by turning the locking cams up.
6
Insert the cassette into the scanner as described in 3.2.3 Inserting a cassette
in the User Manual.
28-4047-23 Addendum to Ettan DIGE Imager User Manual Edition AB
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Scanning Membranes
2 Set Up a membrane scan
Start the Scanner Software by double-clicking the Ettan™ DIGE Imager icon
on the workstation desktop, or by using the start menu: Start:Programs/GE
Healthcare/Ettan DIGE Imager. The scanner software Ettan DIGE Imager window
opens in Setup View.
1. The Tool Bar
2. The Navigation Bar
4. The Setup Area
3. The Information Area
5. The Grid Area
Fig 1. The Ettan DIGE Imager Setup View
2
1
The Tool Bar is used to manage scan templates and to turn on the scanner
light.
2
The Navigation Bar shows the status of the scan workflow.
3
The Information Area shows the instrument status, displays information
about the scan, and includes the SCAN button that is used to start a scan.
4
The Setup Area contains the parameters and settings for the scan.
5
The Grid Area defines the area or areas to scan. The grid coordinates are
aligned with those on the cassette.
28-4047-23 Addendum to Ettan DIGE Imager User Manual Edition AB
Scanning Membranes
To set up a scan
In the Setup Area of the Setup View, the parameters and settings for the scan are
set.
Fig 2. Parameters and settings in the Setup Area
Matrix type:
Choose membrane from the Matrix Type list.
The steps below do not have to be performed in a strict order.
1 Gel format
The Gel format list is used to set the scan area. In the Gel Format list in the Setup
Area choose User Select. In this mode, you can manually set the area to be
scanned. Then use the mouse and drag a box from the positions of the
membrane boundaries. The scan area is shown as a blue box in the Grid Area.
Fig 3. Grid Area
28-4047-23 Addendum to Ettan DIGE Imager User Manual Edition AB
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Scanning Membranes
2 Chemistry
When scanning membranes the chemistry should be Unassigned.
3 Pixel Size
Select the pixel size, i.e. resolution to use, in the Pixel size list. 100 μm should be
used.
4 Channels and dyes
Select the number of channels to be scanned by clicking the Channel check box.
Between one and four channels can be programmed. Selection of the Channel
check boxes must be performed in sequential numerical order. For each selected
channel, select a dye to be scanned in that channel. When selecting dye, the
excitation and emission filter to be used for that dye are automatically presented
by the software. The excitation and emission filter combinations have been
selected to give the optimum results with minimal crosstalk.
Fig 4. Filter settings
5 Exposure
Select the exposure to be used for each channel in the Exposure list. Generally
low exposures are used for membranes. However, a quick test scan on a small
area should be performed to identify an optimal exposure. See 3.4.1 Finding the
optimal settings in the User Manual.
6 Comments
It is optional to add comments to your data. To add comments to the data:
1
On the bottom of the Setup Area, click Experiment Comments to open the
Experiment Comments dialog.
2
Type in the Membrane ID, the Individual Source, and the Sample ID.
3
Type in Comments for the scan. To use the same comments for all the gels
in this scan, select Use this Comment for all Gels. (This option only applies to
the Comments field. All other fields are always specific to each gel area.)
4
Click Close.
To start the scan and for more information see 3.4.2 Start a scan and following
chapters in the User Manual.
4
28-4047-23 Addendum to Ettan DIGE Imager User Manual Edition AB
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