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I hope someone out there can help. My problem is that I want to use the
confocal microscope (BX50 with Fluoview 300) to perform stereological
counts on hippocampal neurons. I am using fluorescence to double label
various markers (NeuN, GFAP, MAP2 with BrdU). First I have to confirm co-
localization (with use of the confocal) and then I would like to use
stereology to count these cells in the hippocampus.

Our lab has just purchased a Microbrightfield system that includes the
Confocal Module. Therefore, if I can take stacks from the confocal
microscope that are sampled in an unbiased fashion then I should be able to
do my counts very quickly.

HOWEVER, how to I sample my areas of interest on the confocal in an
unbiased fashion and then input this information into the confocal module
module that accurate counts can be confirmed.

The confocal microscope has Image Pro Plus 4.5 software.

Basically, my question are:

1. Is it possible to do stereology on cofocal stacks?

2. How do I take images from confocal microscope and apply the
Microbrightfield Confocal Module to do my cell counts?

3. What steps/information would I have to take in order to do the counts
accurately?


Please contact both the listserv and my e-mail directly.

David Laidley (Master's Student)
Memorial University of Newfoundland, Canada

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