Re: paraffin processing of murine embryos

From:Lisa Dillard-Telm

Hi Teri,

I've processed many mouse embryos and they are quite delicate.  Use a plastic or glass pipette with
the tip cut off to gently "suck" up the embryos and transfer them to the next solution.  This method is tedious, but when sample preservation is important you may want to try it.

I fixed the embryos in 4% Para. overnight in the refrigerator with slow rotation in glass
scintillation vials. You do want some agitation so the embryos are properly processed and don't just sit on the bottom of the vial.

The abbreviated processing times are o.k.

I hope this helps,

Lisa Dillard-Telm

"Johnson, Teri" wrote:

> I have 9 and day 10 embryos which need to be paraffin processed.  They were fixed in 4% PFA in PBS overnight (probably overkill) on a rotator (probably too rough).  After an abbreviated processing schedule (roughly 15-20 minutes per station using 1x 70%, 5x Pro-soft, 3x Pro-Par, and 4x Type 6 paraffin) I opened the tissue cassettes to embed them and found them fragmented.
> I need some tips on fixation, processing, and handling these delicate specimens.
> Thanks in advance!
> Teri Johnson
> Manager Histology Core Facility
> Stowers Institute for Medical Research
> 1000 E. 50th St.
> Kansas City, Missouri  64110

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