[Histonet] Staining frozen sections without fixation

Tony Henwood AnthonyH <@t> chw.edu.au
Tue Feb 6 16:37:36 CST 2007


Air-drying frozen sections is a form of fixation. Acetone is a poor
fixative. It probably has some action by dehydrating tissues. It may
also aid in the removal of fat and other hydrophobic cellular
components, thus aiding access of hydrophilic antibodies.

I would not leave sections to air-dry so long, 30 mins would suffice and
I would not bother fixing after the IPX procedure. Morphology will not


Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Martin
Sent: Wednesday, 7 February 2007 3:46 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Staining frozen sections without fixation

Hi All,
I'm trying to get an aPDCA-1 antibody to work on frozen mouse spleen
sections. The antibody is normally used for FACS however the company
says that it works for immunhistochemistry. They recommend staining
before fixation. Usually I cut frozen sections, dry overnight (room
temp) and fix in acetone (10min at room temp) before putting the
sections in PBS and continuing with staining procedure.
If I stain before fixation;
Will the sections survive?
When should I fix - after primary, secondary etc?
What should I fix with?
Any help greatly appreciated.

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