[Histonet] imaging organ cultures

[gayle callis]

Alice, 

You didn't say if your confocal was using an inverted or upright microscope
setup.  If inverted, there are special round glass bottom dishes with cover
glass thicknesses, different diameters designed for viewing with an inverted
setup. They probably work for upright microscopes too. BD Bioscience had
culture plates with very small diameter wells (for inverted microscope)/
Try Mattek (just Google for website) and www.willcowells.com   There were
samples of WillCo Wells at the NSH Symposium/Convention, with both clear or
black glass .  ,  There are even special culture well plates for confocal
use with the well diameters very small.  I believe the culture wells come
from BD Bioscience, and Mattek has the glass bottom petri dishes. Using a
hard set antifade may help anchor your embryonic organ cultures, but these
dishes also come sterile so you can do the cultures in the dish itself,
followed by staining and coverslipping.  

Hope this helps,

Gayle M. Callis
HTL/HT/MT(ASCP)
Bozeman MT 59715 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Alice
Fleming
Sent: Wednesday, October 28, 2009 4:34 PM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] imaging organ cultures

Hi,
I am doing whole mount IHCs on embryonic organ cultures and imaging them on
a confocal microscope.  The IHCs are going well, but I'm having trouble
making the tissue stay in a specific position for imaging.  I have been
putting the tissue in the wells of depression slides, adding about 30uL of
Vectashield mounting medium, then coverslipping.  This works fine, except
that I have little control over the final position of the tissue-as I put
the coverslip down, the tissue often moves.
Any suggestions would be welcome!  Is there, for example, some kind of
optically neutral agarose that I could spot around the tissue before adding
the Vectashield and coverslip??  Or-other ideas??
Thanks,
Alice

Alice Fleming
Department of Human Genetics
University of California Los Angeles
Los Angeles, CA 90095
310-267-2456





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