clearing and mounting after in situ hybridization

[kollmar at rockvax.rockefeller.edu]

Dear Fellow Daniologists,

I have two questions about the best way(s) to prepare wholemounts of
zebrafish larvae for photography after in situ hybridization:

1.  How do you RELIABLY inactivate the alkaline phosphatase once the color
reaction has gone far enough?  Even after washing in TE or PBS with
Tween-20, we observe subsequently a diffuse staining everywhere, including
the yolk, that obscures the actual signal.

2.  When do you choose glycerol, benzylbenzoate/benzylalcohol, or
methylcellulose to mount for photography?

Thank you for any suggestions,

Richard


Richard Kollmar, Ph.D.
Laboratory of Sensory Neuroscience
The Rockefeller University
1230 York Ave., Box 314
New York, NY 10021-6399
Tel. (212) 327 7353
FAX  (212) 327 7352