Posted by
Joel Sheffield on
Nov 16, 2008; 11:32pm
URL: http://imagej.273.s1.nabble.com/counting-cells-in-multiclour-fluorescence-images-tp3694463p3694467.html
This is a particularly interesting problem, since in a single 3-color
image, the yellow arises from the combination of red and green.
Thus, it is possible to separate the color channels to get a red
signal, a green one, and the blue one. However, both the red and the
green channels also carry the information for the yellow cells.
Here's a thought:
You can subtract green from red to get those cells that are only red,
and not yellow. Similarly, you can subtract red from green to get
those that are only green.
Then, you can add red and green to get an enhancement of the yellow
signal.
If your signals are consistent, you can do this in a macro, and then
assign threshold values, and use the analyze particles routine.
Best of luck.
Joel
Date sent: Sun, 16 Nov 2008 12:58:33 -0800
Send reply to: ImageJ Interest Group <
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From: Matthias Kirsch <
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Subject: counting cells in multiclour fluorescence images
To:
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> Dear all,
>
> I am trying to find a way to count cells expressing one or the other or both
> proteins in cultured cells (see e.g. the attached example). There are green
> cells, red cells and yellow cells and I would like to have a
> (semi)-automatic routine with which this can be done objectively. Basically
> the cells can be identified by their nuclei (in the blue channel), but I do
> not know how to count only the nuclei which have a red, green or yellow
> cytoplasm associated with them. As there probably is no way to reliably find
> the outline of a cell with any accuracy (however, maybe maybe one has an
> ingenious approach), it may be sufficient to look in the immediate vicinity
> of the nuclei (e.g. in a ring some pixels wide around the nuclei -user
> selectable preferrentially), if this area is red, green or yellow.
> I would appreciate if someone could help to solve this problem.
>
> Thanx
> Matthias
http://n2.nabble.com/file/n1507033/cells.jpg
>
>
>
> --
> View this message in context:
http://n2.nabble.com/counting-cells-in-multiclour-fluorescence-images-tp1507033p1507033.html> Sent from the ImageJ mailing list archive at Nabble.com.
Joel B. Sheffield, Ph.D
Department of Biology
Temple University
Philadelphia, PA 19122
Voice: 215 204 8839
e-mail:
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URL:
http://astro.temple.edu/~jbs