counting cells with a membrane marker

[eroy]

I have images of T cells in lymph nodes or tumors, stained with an anti-CD8 antibody and another membrane protein (fluorescent, red or green), so the fluorescence generally forms an annulus around the DAPI stained nucleus (blue). When the cells are closely packed as in the lymph node, the membrane fluorescence may be nearly continuous. If I define a cell by its DAPI stain, I'd like to threshold on an annular region around each nucleus since this is where the membranes are. The images are typically at 10x-40x. It seems that surely this has been done before, but I can't find it. Can anyone direct me to a discussion of a similar problem?
Thanks
Ed Roy
University of Illinois at Urbana-Champaign

[rona baron]

Hi,

I can recommend "Cell profiler" software (free on-line), also you can try
and image at x60 and do not do z-stack image...for the analysis use only
one plane..

Rona

On Thu, Mar 8, 2012 at 3:37 AM, eroy <[hidden email]> wrote:

> I have images of T cells in lymph nodes or tumors, stained with an anti-CD8
> antibody and another membrane protein (fluorescent, red or green), so the
> fluorescence generally forms an annulus around the DAPI stained nucleus
> (blue). When the cells are closely packed as in the lymph node, the
> membrane
> fluorescence may be nearly continuous. If I define a cell by its DAPI
> stain,
> I'd like to threshold on an annular region around each nucleus since this
> is
> where the membranes are. The images are typically at 10x-40x. It seems that
> surely this has been done before, but I can't find it. Can anyone direct me
> to a discussion of a similar problem?
> Thanks
> Ed Roy
> University of Illinois at Urbana-Champaign
>
> http://imagej.1557.n6.nabble.com/file/n4557067/2674-2_Irr_sec2_LN1_40x_red_%5B2%5D.jpg
>
> --
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>