[Histonet] RE: Wrinkles on Glutaraldehyde Fixed Tissue

Wed May 7 09:13:59 CDT 2014

Why do you use glutaraldehyde for eyes? Davidson's is a much better fixative for eyes. With Davidson's you have two fast penetrating ingredients and one slow acting fixative. Glut is only a fast fixative. Besides, the best institutions who deal with eyes only use Davidson's. Just my thoughts. 

Bea

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 Ricardo Leyva
Sent: Tuesday, May 06, 2014 11:38 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Wrinkles on Glutaraldehyde Fixed Tissue

Dear Histonetter,

I am having an issue getting rid of wrinkles on sections from 4% glutaraldehyde fixed paraffin embedded ocular tissue. The laboratory's tissue of interest is the retina, and I use glut to avoid retinal detachment (this is mainly on tissue that won't be immunostained). I normally section at 5µm, and fix the tissue 1 or 2 days. I set the water bath at a temperature that is 5-10 C below the wax melting point. I have tried adding ethyl alcohol to the water and the wrinkles remain-or takes very long for most of them to be gone. I have tried placing the sections  in a 1:15 mix of ethanol and water-and have also tried 30% ethanol in water, both at RT, before placing the sections on the water bath; when I do this, the sections separate abruptly (once in the water bath) from the slide I use for transferring the tissue, causing tissue layer separation. When I work with 10% formalin, the section wrinkles disappear nicely in the water bath that contains water and a bit ethanol. Nonetheless, when it comes to 4% glut it just does not work.

Any input in troubleshooting this is greatly appreciated.

Best,

Ricardo Leyva
(858) 822-1629
rleyva <@t> ucsd.edu
UCSD Shiley Eye Center

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