[Histonet] fixing sponge larvae for in situs

From:Gary Radice

I have a colleague who wants to do in situ hybridizations on sponge  
larvae  (yes, sponges are animals and they have larval forms!). She  
tried fixing some in 4% paraformaldehyde in sea water, and the larvae  
exploded. She also tried Carnoy's fix, and 4% PFA in Millonig's  
phosphate buffer, and the same thing happened.

The only thing I could think of to suggest was dropping to 1 or 2%  
PFA in sea water. I've seen references to using Bouins for light  
microscopy or osmium tetroxide for TEM, but I don't think either of  
these approaches would be good for in situs.

Any other ideas?

Gary P. Radice			gradice@richmond.edu
Department of Biology	804-289-8107 (voice)
University of Richmond	804-289-8233 (FAX)
Richmond VA 23173	http://www.richmond.edu/~gradice

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