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Given that each astrocyte has a single nucleus and assuming that you can arrange
to mark all nuclei, you then need to count nuclei (much easier to segment) that
are within an astrocyte (however defined).  This does not require that you can
fully demarcate the limits of the astrocyte, but only to make a yes/no statement
for the area around each nucleus.

The next issue is obtaining a count that is independant of cell size - you can't
simply count the number of nuclei in a single 2D image- the best ploy is to use
sequential sections and look for the appearance or disapperance (either but not
both) of nuclei which will only occur once per nucleus. Producing a count for a
given volume.  This may suffice but you may actually want an estimate of the
total population in each brain or brain region, which requires that this volume
be measured.

look at the stereology literature.

J. Adler
NIMR
London

Mike Goldsworthy wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>      I have a researcher here who wants to count astrocytes in brain tissue.
> We are using Image Pro Plus and have tried segmenting out the cells, but
> because of their unusual and varied shapes, we are not getting accurate
> results. Is there some feature that we can use to isolate them and do a
> count. Can anyone out there help us. Thanks in advance.
>
> Michael Goldsworthy
> Confocal Laboratory Supervisor
> Faculty of Medicine, HSC
> Memorial University
> St. John's, NL
> A1B 3V6
> (709) 777-8470