Hi everyone,
I have a problem with FIJI for analyzing my ICC results.
My images have three channels: blue, green, and red. I want to measure and compare the fluorescent intensity of green and red channels. I planned to split the raw image into three single channels and convert them into grey-scale images by clicking on Image -> Type -> 8-bit. I will then measure the Corrected Integrated Density (CTCF) of each single channel by the following formula:
(CTCF) = Integrated Density - (Area of selected cell X Mean fluorescence of background readings)
The problem is that I could not convert the RGB images into the grey images by that way. I returned to ImageJ but only 1 or 2 channels were converted.
Is there anyone having the same problem? How did you solve it? Or any suggestions to measure the fluorescent intensity?
Thank you.
if you have RGB format image, using ImageJ is pretty straight forward for splitting the Channels. When you open your ICC results, is ImageJ reported as at least 24 bit Image? an example of your Image would be help full to try in ImageJ.
If the image needs to be split, use "Image, color, split channels." Then convert to greyscale using "Image, type, 8 bit."
Most imaging software will allow you to save the data in the greyscale, monochrome format of the detectors. Unless you are suing a color camera, I don't understand why your data is getting converted to RGB in the first place.
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