[Histonet] IHC general fixation

[Morken, Timothy]

In general, since the advent of heat-mediated antigen retrieval long fixation time is not an issue. In fact there are more problems associated with short fixation times (less than 6 hours) and IHC, than long fixation times. Studies have shown that even tissue left in formalin for weeks, months, even a year, will have good antigenicity with heat antigen retrieval.

The key for good reproducibility is to make your fixation times consistent, not wildly different from day to day and embed rather than keeping in other solutions like alcohol.

Tim Morken
Supervisor, Histology, Electron Microscopy and Neuromuscular Special Studies UC San Francisco Medical Center San Francisco, CA

Tim Morken
Supervisor, Histology, Electron Microscopy and Neuromuscular Special Studies
UC San Francisco Medical Center
San Francisco, CA

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of William J. O'Connor III
Sent: Wednesday, November 12, 2014 9:56 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] IHC general fixation



I'm looking for input on the latest theories of duration of fixation for routine IHC.  In the past, it was thought that tissues should be removed from 10% NBF after a max of 72 hours,  and if not immediately processed, transferred to 70% ethanol until processed to retain antigenicity.  What is the current thinking/trends?  Over the last couple of years, I have heard that leaving tissues in 70% ethanol may be doing more harm than good, and it's NOT a good idea to leave them in alcohol for extended periods of time.  I would appreciate your input, or point me towards any new data.

Jackie O'


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