AW: [Histonet] sections falling off in day 2 ISH

[Gudrun Lang]

Hi
I do FISH on 2-3 µm human paraffin sections. They are mounted on Superfrost
Plus slides, dried at least over night in an 56°C oven, sometimes over the
weekend without negative influence.
Hybridization is done in a Hybridizer, coverglass sealed with rubbergum over
night. Next day I put away the rubbergum and the coverglass with care, just
lifting the coverglass with a scalpell or needle.
Then put the slides directly in the prewarmed stringent-wash-buffer (here
55°C 5 min), rinse in dest. water, let airdry and coverslip with DAPI
contenting mounting medium.

You may compare your protocol to mine. We hardly had problems with falling
off sections. One time we even had success to stain a section on a "normal"
glass slide, but treated it like a raw egg.

Bye
Gudrun

-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Megha
Kumar
Gesendet: Freitag, 10. August 2012 11:11
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] sections falling off in day 2 ISH

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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