SPAM-LOW: [Histonet] cryosection for IHC samples

[Patsy Ruegg]

It depends on the marker you are using.  Some antibodies require
acetone/ethanol fixation and are not recommended to be used with aldehyde
fixatives.  If you know your marker works with PFA fixative why not fix it
and then process it into paraffin.  I use frozen sections just for tissues
and markers that may not do well with aldehyde fixation and paraffin
processing.

I use fixed tissues that are then infiltrated in 30% sucrose mostly for
tissues that will be difficult to cut, such as calcified bone and then use a
tape transfer sectioning technique.  Since you are working with tumor you
probably do not need to fix and then cyroprotect in sucrose as the unfixed
snap frozen sections should section pretty well directly.  Decide from there
if you need to fix in acetone or PFA (depending on what ab you are using).

Regards,
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 Carolina
Soekmadji
Sent: Saturday, September 04, 2010 6:14 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: SPAM-LOW: [Histonet] cryosection for IHC samples

Hi,
Could anybody give me an advice about sample processing for cryosection? For
best IHC, is it better to go with fresh tumour sample that goes straight to
cryosection, then fix with PFA ? How long we need to fix in PFA in this
case? 30 min?
Or is it better to process it by fixing tumour first by immersion in PFA
then sucrose then cryosection ?

Best wishes,

Carolina


Carolina Soekmadji, PhD. MSc.
Postdoctoral Research Fellow
Australian Prostate Cancer Research Centre - Queensland | Institute of
Health and Biomedical Innovation -QUT |
t: 07 3176 3099 (APCRC - Q, Princess Alexandra Hospital)
t: 07 3138 6286 (IHBI, QUT Kelvin Grove)
mobile: +61 423 111 807
f: 07 3176 7440
e: carolina.soekmadji <@t> qut.edu.au<mailto:carolina.soekmadji <@t> qut.edu.au>


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