shrinking tissue

From:"Dr. Maneesha.S.Inamdar"


Hi,

	I'm trying to embed mouse embryos (upto d10.5)
in paraffin for sectioning.  Often when the embryos
are transferred to 50% wax after xylene, they
shrink to almost half their size and are not
worth embedding.  This happens with stained as
well as unstained embryos. Can anyone tell me what the
cause might be and how to prevent it?

	I looked up several protocols and websites and
none talk of tissue shrinking! So may be I'm just making
some basic mistake? I am new to paraffin 
sectioning and we don't have an embedding centre/station. 
I melt the wax in an oven at 58 degrees and do all the
operations for embedding on a heating block at 58
degrees.  I use paraffin wax from Merck or
Ranbaxy that has a melting temp. of 58-60 degrees.

	I would appreciate any comments/help
from histonetters.  Thanks a lot.

Maneesha Inamdar
Molecular Biology and Genetics Unit
JNCASR, Jakkur
Bangalore, INDIA





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