Whole Mount In Situ Problem

vilimf01 at mcrcr6.med.nyu.edu vilimf01 at mcrcr6.med.nyu.edu
Tue Dec 16 08:26:52 EST 1997


Hi all,

   I am trying to do whole mount in situ hybridization on marine
mollusc ganglia.  Everything is fine until I get to the hybridization
step when the tissue invariably shrinks and becomes impenetrable.  I
have tried many ways to prevent the shrinkage and maintain access
including varying the detergents, proteinase K, boiling in citrate
buffer, changing the hyb buffer and conditions.  Bottom line is that
any denaturing conditions which are needed for the hyb cause the
shrinkage.  Any ideas about how to get around this problem? I am at a
loss.  I have tried many of the protocols people use for vertebrate
whole mounts of embryos and such without much success.  TIA

							Best Regards
							SVEN



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