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8.3.2
To Process a Blot
1.
Place the Western blot on a hinged transparency or in clear plastic bag.
2.
Position the blot on the black chemi tray.
3.
Pre-focus and position the blot with the lights off, the door cracked open and the lens
set to the fastest f-
stop (the lowest aperture number). Use the “Chemi Blot Pre-Capture
Positioning” template discussed earlier.
4.
Apply the chemi substrate on the blot (front side up). Then, close the hinged transpar-
ency over the blot and smooth out any bubbles. Reposition the blot in the field of view if
needed. Close the darkroom door.
5.
Immediately start previewing with the “Chemi Blot with Fast Capture and Lower Resolu-
tion” template. Let the blot develop inside the darkroom while watching the preview.
When approaching maximum signal (which should take 2 to 3 minutes) use the “Chemi
Blot with Lower Capture and Higher Resolution” template to capture the image.
6.
Light emission will drop off quickly, so capture several images as quickly as possible.
7.
Save and archive original files for future quantitative analysis.
8.
After creating a copy of an image file, use the image invert function in VisionWorks,
adjust the image histogram for optimal viewing, and burn the changes to a new file.
IMPORTANT
Capture and image processing steps are described in more detail in the software manual and in the
software help files.
