[Histonet] sections falling off in day 2 ISH (Megha Kumar)

[Therese C. Thinnes]

Hi Megha, Not sure why your sections are falling off. When I did ISH I  used  Colorfrost Plus Slides. I would cut at 4 microns-mouse or human.  I would dry them at room temp a few hours followed by 55 C for an hour. I would use the slides the next day or next week, never the same day. On day 2 my SSC was 60 C and I used a stir bar for 2 hours on slow stirring. Rarely did I lose any sections.
Terri Thinnes
Research Assistant
The Scripps Research Institute
La Jolla, CA




Message: 13
Date: Fri, 10 Aug 2012 14:40:30 +0530
From: Megha Kumar <meghak <@t> g.clemson.edu>
Subject: [Histonet] sections falling off in day 2 ISH
To: histonet <@t> lists.utsouthwestern.edu
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Hello everyone!
When I subject murine skin and intestine paraffin sections (7-10microns) to
ISH protocol, the sections fall off on day 2 when I add the SSC. The
sections are taken on ploy lysine coated slides and remain on the slides
when I do other protocols such as antibody staining. Can anyone suggest me
why this is happening? How to prevent the slides from falling off on day 2?
Thank you for all the help!
regards
Megha



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