[Histonet] mouse embryo section tears
sladd <@t> hsc.usf.edu
Mon Dec 15 10:16:59 CST 2003
I just did a bunch of mouse embryos (and placentas). I asked the
researcher to fix them in Bouins fixative ( for less than 24 hours)
since I had read that this was the best (non-mercury) fixative for
embryos. I also asked that the researcher start dehydration at 30%
alcohol and give them to me in 70% alcohol. The embryos cut like butter.
Bouins fix will lyse red blood cells and those red cells/blood might be
what is causing you problems in the liver area? I wonder if what you are
calling tears is actually "chatter"? Are you soaking the block in ice
water before sectioning? This might alleviate your problem. I also cut
mine at 7 microns but I didn't have to soak the block and I did not have
any problems with chatter. Obviously, going back and changing the
fixative is not an option, but maybe next time you could try Bouins?
University of South Florida
Rashmi Chandra wrote:
> I am sectioning paraffin embedded mouse embryos (fixed in 4%
> paraformaldehyde) at 7 microns. The sections cut and ribbon fine but
> after i dry them on the slide, tears appear in the liver. These tears
> are not present when I first check the sections after lifting them
> from the water bath (temperature 37 C) onto the slide. I think the
> tears may result from differential drying of the various tissues, and
> I am looking for a way to prevent them.
> Thanks a lot for your advice!
> Rashmi Chandra, Ph.D.
> Dept. of Neurobiology
> Duke University Medical Center
> Durham, NC 27710
> Tel: (919) 681-5449
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
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