[Histonet] Please Help!

[Tony Henwood]

Nick,

NBF may not work for Delia. She needs to do IF on the frozen sections
and NBF fixation will require her to do AR on the sections (esp if she
is localising immunoglobulins and complement).

Possibly warming the frozen tissue, using a thumb, may help.

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Nicholas
David Evans
Sent: Saturday, 1 August 2009 5:37 AM
To: 'DELIA GARCIA'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Please Help!


We fix skin by sandwiching it between two sponges soaked in either 4%
PFA or 0.4%PFA (for paraffin or cryo respectively) and compressed in a
cassette. This flattens the skin. Following several hours fixation the
skin is usually flat and rigid enough to handle and embed. It can then
either by dehydrated for paraffin or soaked in sucrose for cryo.

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of DELIA
GARCIA
Sent: Friday, July 31, 2009 11:42 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Please Help!

Hello,

I am in dire need of some suggestions. I work for a Derm lab and we
occasionally get fresh skin punches that we freeze to do FITC
(immunoflouresence). I have a really hard tiime keeping the tissue from
curling makeing almost impossible to get a decent section on the slide.
Anyone out there that might have experienced the same thing I would love
to learn your techniques that overcame this problem. I have tried
lightly blowing, freeze spraying, and lightly brushing acetone on the
cold plate (not all at the same time). Nothing seems to ease it up.
PLEASE HELP! I have one in the cryostat now. Thank you so very much.

Delia



      
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