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Photoshop Procedure to Colorize Immuno Images
Open Photoshop
Open both images (Cy3-red and Hoechest33258-blue)¡ª
1. Choose one image
Image¡ªMode¡ªRGB Color
Image¡ªAdjust¡ªBrightness/Contrast.
Then Save As:
2. Choose other image
Image¡ªMode¡ªRGB Color
Image¡ªAdjust¡ªBrightness/Contrast.
Then Save As:
3. Select one of the images¡ª(say the Hoechest33258-blue)
Go to Channels
Click on Red¡ªGo to Image¡ªAdjust¡ªBrightness/Contrast
Move both sliders to zero (or -100)
Click on Green--Go to Image¡ªAdjust¡ªBrightness/Contrast
Move both sliders to zero (or -100)
Save
4. Select the other image (Cy3-red)
Go to Channels
Click on Green¡ªGo to Image¡ªAdjust¡ªBrightness/Contrast
Move both sliders to zero (or -100)
Click on Blue--Go to Image¡ªAdjust¡ªBrightness/Contrast
Move both sliders to zero (or -100)
Save
5. Select one of the images
Go to Layers
Double click on Background¡ªRename: Cy3-red
Save
6. Select the other image
Go to Layers
Double click on Background¡ªRename: Hoechest33258-blue
Save
7. Select All
Copy
8. Go to the other image
Paste
Change from ¡°Normal¡± to ¡°Screen¡± on Layers pallette.
Save As
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