利用PHOTOSHOP将两种荧光图象重叠，从而达到激光共聚焦效果，可以节省不少

操作步骤如下：

Photoshop Procedure to Colorize Immuno Images
Open Photoshop
Open both images (Cy3-red and Hoechest33258-blue)

1. Choose one image
Image—Mode—RGB Color
Image—Adjust—Brightness/Contrast.
Then Save As:

2. Choose other image
Image—Mode—RGB Color
Image—Adjust—Brightness/Contrast.
Then Save As:

3. Select one of the images—(say the Hoechest33258-blue)
Go to Channels
Click on Red—Go to Image—Adjust—Brightness/Contrast
Move both sliders to zero (or -100)
Click on Green—Go to Image—Adjust—Brightness/Contrast
Move both sliders to zero (or -100)
Save

4. Select the other image (Cy3-red)
Go to Channels
Click on Green—Go to Image—Adjust—Brightness/Contrast
Move both sliders to zero (or -100)
Click on Blue—Go to Image—Adjust—Brightness/Contrast
Move both sliders to zero (or -100)
Save

5. Select one of the images
Go to Layers
Double click on Background—Rename: Cy3-red
Save

6. Select the other image
Go to Layers
Double click on Background—Rename: Hoechest33258-blue
Save

7. Select All
Copy

8. Go to the other image
Paste
Change from “Normal” to “screen” on Layers PALLette.
Save As: