[Histonet] Which fixative for eyes

[Bea DeBrosse-Serra]

Davidson's fixative.

Beatrice DeBrosse-Serra HT(ASCP)QIHC
Isis Pharmaceuticals
Antisense Drug Discovery
2855 Gazelle Ct.
Carlsbad, CA 92010
760-603-2371




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Elizabeth Cameron
Sent: Wednesday, January 16, 2013 11:49 AM
To: Lucie Guernsey; Louro, Pedro
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Which fixative for eyes

We work with mice and do a variety of fixatives, but we prefer a methanol/acetic acid fixative.  It holds the shape of the eye very well, although it may not show the Schlemm's canal as well as other fixatives.
-Liz

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Lucie Guernsey
Sent: Wednesday, January 16, 2013 1:59 PM
To: Louro, Pedro
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Which fixative for eyes

I'm curious about this as well.

We work with mice and rats in our lab and whenever we need eyes (corneas, more specifically), we immersion fix them whole in Bouin's solution overnight at 4C then transfer to 70% EtOH until we're ready to excise the corneas, process and embed into paraffin. We've found that we get nice half-moon sections with good looking stromal and epithelial layers this way compared to fixing the eyes in 4% PFA (which yielded squiggly, damaged corneas). We then perform IHC.

Would love to know if there's a better way, even though this seems to work for us.

Thanks,
Lucie

Lucie Guernsey
UCSD
lguernsey <@t> ucsd.edu



On Wed, Jan 16, 2013 at 10:32 AM, Louro, Pedro < LouroP <@t> princeton.huntingdon.com> wrote:

> Hello to the eye experts,
>
>
>
> I am currently working on a large animal (LA) eye project which will 
> give me the flexibility of experimenting with different fixatives.
>
> I am currently using Modified Davidson's solution (48 hours) followed 
> by a 10% NBF solution prior to trimming the eyes, embedding , 
> microtomy and H&E staining.
>
>
>
> After literature research, I found that different labs are using:
>
>
>
> Modified Davidson's solution
>
> Davidson's solution
>
> 10% NBF
>
> Gluteraldehyde
>
> Some places inject the eye with fixative before submerging the eye 
> itself.
>
>
>
> I wanted to get some ideas / preferences from the experts out there so 
> I can cover my entire basis before finalizing my conclusions.
>
>
>
> Thanks in advance to all,
>
>
>
> Pedro Louro
>
> Group Leader Histology / IHC
>
>
>
>
>
>
>
>
>
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