http://imagej.273.s1.nabble.com/Segmentation-of-confluent-cells-tp5007785p5007808.html
If you stain the nucleus and the cell membrane - there are a number of fluorescent stains for the cell membrane- then locate the nucleus of each cell and use a region growing algorithm until you encounter the cell membrane of that cell. Since the cells are varied in shape you will have to do this in various directions from the nucleus. There is a program that does this but is very expensive for a license - it is eCognition. If you or someone in your group understands image processing and analysis you could probably find references to region growing algorithms and write one within ImageJ.
I would try a membrane marker : wheat germ agglutinins or DiD lypophilic stain (works with living cells...)
> Hi all,
>
> I've been asked whether its possible to measure the area of individual
> cells in a confluent monolayer autonomously. As far as i'm aware this
> would be near impossible with brightfield techniques so i have
> suggested the cells are fixed and stained but i was wondering which
> structures would be best to label to give us the best chance of being
> able to do the analysis using ImageJ. I was thinking the nucleus and
> some kind of cell junction marker would be best. I'm not sure a
> cytoplasmic marker would add much if the cells are all touching.
>
> As far as analysis goes, my knowledge of segmentation and separating
> objects goes as far as the basic thresholding and morphological
> operators erosion, dilation, watershed etc. Are there any good
> specific tools/plugins for this kind of work? And are there other
> more advanced methods of segmentation of separating touching objects
> available? Perhaps something like level sets would be good for this? I
> think the basis of my approach would be nucleus identification to
> pinpoint a single cell, cell junction enhancement and final segmentation of some kind.
>
> Thanks for the advice,
>
> Matt
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