Koshland Web Site/Methods
IMAGE QUANT QUANTITATION
In ImageQuant Program:
  1. Change monitor to 256 colors via the control strip
  2. Open ImageQuant and open file
  3. Change contrast on toolbar (contrast button is the rectangle with a diagonal line through it). Set range, put hi setting at 100
  4. Pick the selection box on left (this is the rectangle with a filled square at each corner) If necessary enlarge image by picking 'zoom in' from new menu
  5. Draw rectangle around band
  6. Pick the arrow pointing
  7. Select rectangle by putting cursor arrow on it. Hold the mouse down, move rectangle to second band, hit V. Move rectangle to third band, hit V. Continue until all desired bands have been selected
  8. Move rectangle to blank area for background, hit V
  9. Put rectangle 1 on first band
  10. Go to analysis; choose background correction
  11. Select all rectangles
  12. Pick object average and pick background rectangle, close
  13. Go to analysis; choose volume setup, display OK
  14. Go to analysis; choose volume report, click display
  15. Name excel file and save
  16. Check values to make sure they approximate image (remove contrast first by resetting hi to 25%) Negative values can result from differences in background across gel
  17. Save as microsoft excel workbook, hit replace

Microsoft Excel processing:

  1. Move totals so they are next to relevant chip, first copy group, second paste into next column (usually C)
  2. Type formula into D2 (=b2*100/(c2*90)
  3. Click on the check mark above window
  4. Click on D2
  5. Copy D2
  6. Select boxes below C2 corresponding to chips
  7. Paste
  8. Pick number in category decimal places 2
  9. OK
  10. Save
Comments:
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