[Histonet] IHC on fresh frozen

Tony Henwood AnthonyH <@t> chw.edu.au
Thu Dec 4 15:56:04 CST 2008

Do you roll it in the paraformaldehyde powder?


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 Andrea
Sent: Friday, 5 December 2008 3:12 AM
To: FU,DONGTAO; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC on fresh frozen

Try fixing for 5-10 min in PFA.

At 10:16 AM -0500 12/3/08, FU,DONGTAO wrote:
>Hi, all
>  Recently I did some IHC(chromagen methods) on mouse fresh frozen
>tissues, mainly using insulin antibody on pancreas. The image is 
>much fuzzier compare to paraffin embedding tissue. And the staining 
>also smeared to acinar cells which surround the islet.
>  I airdried slide(>30min) and used a general acetone method(-20C
>5min) to fix the tissue before I did IHC.
>  How can I get a relatively sharp staining on the fresh frozen
>tissue?Does anyone here have any experience on it? Any suggestions?
>  Many thanks and have a nice day,
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu


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