| Author: |
Sergio Caballero (ksamn2@ufl.edu) Significant parts of this code were adapted from Wayne Rasband, Pierre Bourdoncle, and Christopher Philip Mauer. |
| History: |
2005/04/01: First version |
| Source: | ColocalizeRGB_.java |
| Installation: | Download ColocalizeRGB_.class to the plugins folder and restart ImageJ. |
| Description: |
This plugin finds the colocalized pixels in three separate channels (red,
green and blue). It was written primarily to analyze 3-color immunofluorescent
microscopy images, although any RGB image may be analyzed. The plugin will
work with either single images or with stacks. Colocalized pixels will be
displayed for Red-Green, Red-Blue, Green-Blue, and Red-Green-Blue
colocalization. The plugin can also generate three separate RGB images which
overlay the colocalized image on its respective analyzed channels - for
example the Red-Green colocalized image is placed in the blue channel along
with the original 8-bit red and 8-bit green images to generate an RGB overlay
image.
To use:
1. Open three 8-bit images (or stacks) each containing separate data for
red, green or blue channels. With a single RGB image use the ImageJ command
Image->Color->RGB Split. Close the blank channel images (e.g., split the
red image, close the green and blue channel windows).
2. Run the plugin and select the appropriate Red, Green and Blue 8-bit
images from the drop-down lists.
3. Auto-thresholding is defaulted. Uncheck the box and enter a threshold
level for each channel if manual thresholding is desired (value = 50 is
default, acceptable values are 0-255).
4. A pixel ratio between paired channels is defaulted to 50%. Check the
appropriate box to enter another ratio (acceptable range is 0-100%).
5. Check the box to show the RGB colocalization overlays if desired.
6. Click OK to run.
|
