UVP ChemiDoc-It Imaging System - Manual (Page 13)

ChemiDoc-It Imaging System 13
Using Templates
Set Up and Save Camera Templates
NOTE: Without using templates, camera settings can be manually adjusted directly in the software.
Chemi Blot Pre-Capture Positioning: For this application, use a preview binning of 1x1, a capture
binning 1x1 and an exposure time of 0.2sec. Have the darkroom door cracked open to let in some
ambient light.
Chemi Blot with Fast Capture and Lower Resolution: For this application, use a preview binning
of 8x8, a capture binning of 4x4, and an exposure time of 15sec. This setting is useful for quick
screening; the time can be increased or decreased depending on the intensity of the signal from the
blot.
For better resolution, set the preview binning to 8x8, the capture binning to 2x2, and the exposure
time to 60sec. The preview is the same and the exposure time is intermediate, but the final image is
of higher resolution than when using 4x4 binning.
Chemi Blot with Slower Capture and Higher Resolution: For this application, use a preview
binning of 8x8, a capture binning of 1x1, and an exposure time of 5min. This will result in the best
possible capture resolution. However, the exposure takes longer because the image is not binned.
Set Up Darkroom Templates
NOTE: Without using templates, darkroom settings can be manually adjusted directly in the software.
Chemi Blots: Place a business card on the chemi tray in the ChemiDoc-It darkroom. Set the lens to
the fastest f-stop (the lowest aperture number) and focus on the business card with darkroom door
cracked open, no emission filter in place, no lighting on, and either the lens zoomed or the lift platform
positioned to fit the blot.
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) and then close the hinged transparency
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 Resolution”
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 VisionWorksLS, adjust the
image histogram for optimal viewing, and burn the changes to a new file.
Note: Capture and image processing steps are described in more detail in the VisionWorksLS
software manual and in the software help files.