[Histonet] embedding protocol for embryos

Chris Pomajzl cpomajzl <@t> cpllabs.com
Thu Aug 17 08:03:50 CDT 2006


When I was in research, we would process very young embryos for RNA ISH as
well. We would fix the embryos while still in the decidua. In fact, at
E8-E9, we would simply cut the uterus between each embryo and have them drop
into normal saline and then transferred to fixative. These were fixed in 4%
PFA overnight in 4'C refrigerator on a rocking platform. Embryos were
transferred to normal saline or PBS the next day. Once the embryos are
harvested from the decidua, they are hand-process with graded ethanols and
xylene. 3-5 minutes is sufficient for each step with slight agitation to
facilitate diffusion. Three paraffin changes at 5-10 minutes followed by
embedding. Hope this helps.

Chris Pomajzl, HTL (ASCP)

Histology Supervisor
Clinical Pathology Laboratories, Inc.
9200 Wall Street
Austin, Texas 78754
512.873.1660 (o)
512.873.5004 (f)
cpomajzl <@t> cpllabs.com

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----- Original Message ----- 
From: "Vicki Hakim" <hakim <@t> bgu.ac.il>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, August 17, 2006 6:26 AM
Subject: [Histonet] embedding protocol for embryos

hi all
does any one know a good protocol for embedding young mouse embryos( E-8/9)?
the embryos are used for mRNA in-situ hybridizations. i have been trying
different washing protocols and the tissue is always porous. if anyone could
comment on fixation time and the time recommended for such small tissue to
be in the melted paraffin that would be great.
vicki       ‎
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