Adobe Image Protocol
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1.    Open Image
2.    Edit Photo

  • Crop Image:
    • Find button on the menu on the left that looks like a picture frame > click it and crop area > Press enter to crop > Locate and click magnified glass on left button menu screen to zoom in
  • Locate adjustment layer button found on lower left quadrant on screen under layers section. Button looks like a moon half full, or one of those black and white cookies. Click button.
  •  If neurons do not stand out...
    • Click on levels.  Adjust input levels. In general white arrow should be moved to left so picture is a little brighter.
    • Still under adjustment layer button click brightness/contrast, Put contrast all the way up. Lower brightness to further contrast.  Also helps to adjust brightness/contrast setting repeatedly.
    • Click Hotkey & E at same time (to merge layers)
  • If neurons do stand out...
    • Click brightness/contrast.  Put contrast all the way up and raise up brightness to enhance contrast
    • Click Hotkey & E at same time (to merge layers)
  1. Edit Photo (Part DUEX)
  • Change Effects
    • Click Filter menu on top > Go down to artistic & Click on Poster Edges effect.  Adjust the 3 meters on left side.  In general put Posterization between 1-2, Edge between 1-3 and thickness between 1-3, but can vary greatly. (Its okay if little fuzzies are on side because will go away during threshold and following steps.
  •  Click adjustment layer button > Click Threshold.  Find best fit.
  • Click Shift, Hotkey & E at same time

•    Use Eraser to delete Nucleus and other erroneous background.  Overall just get of bigger parts b/c little parts will go when photo is analyzed under image J. A good indicator of particles that should be deleted is to set eraser at 10 pixels and delete everything smaller then that, but once you get the hang of that just use the big eraser.

  1. Save Photo
  • Make sure photo is put in separate folder designated for pictures ready to be measured in Image J. Do not go to IMage J until you have a lot of pictures because Image J will run macro that can analyze many areas very quickly.

- This looks complicated but once you get the hang of it, it is much more accurate and easier than Image J for editing the photos.

To measure area of pictures go to child "Image J Macro Area analysis"

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