Hello,

I work with chicken embryo (2-3 day old) whole mount embryos. My interest
is to look at diI and diO labeled myotome fibers in developing somites
using confocal microscopy (Nikon PCM2000). I also need to preserve the
tissue structure. Therefore, I have done this by layering black
electrical tape over a clean glass slide. The desired thickness is up to
you. For my needs anywhere between 3-5 layers of tape works. Then, I cut
out a small square chamber in the tape with a pointed tip razor blade.
The tissue is then placed in the space containing PBS and coversliped. To
make it temporarily sealed, I use nail polish over the coverslip and
black tape. For immunofluorescence work, I can use PBS:glycerol (diluted
1:1) with 2% n-propyl-gallate with the chamber which works fine. My paper
shows examples of my confocal imaged work and describes the chamber and
other conditions used (Denetclaw et al., 1997, Development
124:1601-1610).

Good luck,

Wilfred

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Wilfred Denetclaw Jr., Ph.D.             Tina Denetclaw, Pharm.D. , BCPS
Department of Anatomy and                Assistant Clinical Professor
Cardiovascular Research Institute   &    School of Pharmacy (Box 0622)
University of California                 University of California
San Francisco, CA. 94143-0452            San Francisco, CA  94143
(415)476-1530 (Office)                   [log in to unmask] (E-mail)
(415)476-4845 (FAX)
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