I've taken fluorescence microscopy images with 4 channels (DAPI, FITC, Cy3, and Cy5 - Cy5 pseudocolored white) and now I can't figure out how to separate them for analysis on ImageJ or Photoshop. A quick google search indicates that because the images are RGB, the white channel is forever shown on all channels and can't be separated out. Is there any way to get around this? Thanks in advance!
Hi there,
Unfortunately, there is no solution for your problem. Indeed, the pseudocolored white will be permanently "stuck" to the other channels (by definition of an RGB image). The only solution is to take images again...
Check this in ImageJ forum:
https://forum.image.sc/t/problem-to-split-4-channel-image/6374
As and advice, never save fluorescent microscopy images as RGB, but as the multi-channel TIFF (or any other format that supports multi-channel acquisition and storage).
Sorry to not be able to help,
Cheers,
J
Hi Marisa Adhikusuma Jeffries . I agree with J. Ramirez-Franco . You won’t be able to split an RGB image into 4 channels. You have to save your data as a .tif or the format associated with your imaging system.
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