[Histonet] Drying of fixed cryosections (need to preserve tissuemorphology)

[Patsy Ruegg]

U could put a jar of acetone/ethanol mix in the cryostat and go directly
into the  fix (75ml of acetone and 25mls of 100% ethyl alcohol), u can go
directly to buffer and never airdry your sections, see how that works for
you.

Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net 
www.ihcrg.org

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Keng Ng
Sent: Monday, February 16, 2009 9:26 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Drying of fixed cryosections (need to preserve
tissuemorphology)

Hi, I have a question regarding transport and storage of
glutaraldehyde-fixed cryosections. 

I make cryosections of human skin tissue where good morphology is important
for the intended application. My samples are fixed in 2% glutaraldehyde and
embedded in OCT medium.

However, I've realised that drying cryosections on coated slides (Superfrost
plus) at room temperature makes tissue in the dermis (but not in the
epidermis) separate into layers, with a lot of gaps in between. Whereas
fresh out of the cryostat, the tissue looks fine, solid and tightly packed.
I presume this is to do with formation of ice crystals as the tissue thaws?
If I keep the tissue wet when it's out of the cryostat, that holds off the
damage for a while, but as soon as I let the cryosections dry at room
temperature after that, the same damage happens.

So it would seem that perhaps I need to dry the cryosections differently. I
would be grateful if anyone could point me in the right direction.

Many thanks in advance.

Keng Wooi Ng
Welsh School of Pharmacy
Cardiff University

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