[Histonet] Mice embryo cryocuts

[Mehlika Faire]

Sorry for the  repost- forgot to include everyone else.

Do you fix the embryos pior to sectioning? or post fix? Usually I place my embryos in 30% sucrose over night in the 4 degree (or until they float to the top-this also depends on the age/size of the embryos) and I fix them in 4% PFA for 1 hr to overnight, depending on the age of the embryo or what the type of treatment for IHC (antigen retrieval w/ trypsin or dnase the embryos requires longer fixation).  Then I embed them and section.  I haven't had any problems with this method. Hope this helps you out.-Mehlika
> Date: Wed, 31 Aug 2011 09:20:09 -0400
> From: talulahgosh <@t> gmail.com
> To: Carmen.M.Garcia <@t> uv.es; histonet <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] Mice embryo cryocuts
> CC: 
> 
> You need to leave the embryos in sucrose waaay longer than 5 hours.  I would
> leave them at least overnight, if not more than that.  You can probably take
> them out of the blocks and resink them in sucrose, if you're desparate.
> But next time, always sink (ie wash in sucrose) for at least overnight.  How
> old are the mice you're sectioning?
> Write back if that doesn't help!
> 
> Emily
> 
> A great book should leave you with many experiences, and slightly exhausted.
> You should live several lives while reading it.
> -William Styron
> 
> 
> 
> On Wed, Aug 31, 2011 at 3:37 AM, Carmen Maria Garcia Pascual <
> Carmen.M.Garcia <@t> uv.es> wrote:
> 
> >
> >
> > Good moorning everyone:
> >
> > I am Carmen Garcia from the University of Valencia. I am just starting
> > my PhD. Now I have to obtain mice embryo cryocut and I am having some
> > problems with the morphology... We removed the embryos and put them in
> > sucrose 30% like 5 hours, then embebbed in OCT and put them at -80ºC,
> > when we are going to cut them we let the in the cryostat like 2 hours
> > and the cut them.
> > Someone can give me any advise of what can we been doing wrong?
> > Thank you so much.
> >
> > *******************************************************
> > Carmen María García Pascual
> > FIVI/INCLIVA/Facultad de Medicina
> > Universidad de Valencia
> > Departamento de P.O.G (Laboratorios)
> > 96.386.40.48
> > Avd. Blasco Ibañez 17
> > 46010, Valencia
> > *******************************************************
> >
> >
> >
> >
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