[Histonet] titron vs DMSO

[Rene J Buesa]

Triton is a surfactant used to facilitate the dispersion of  IHC reagents but if you say that without Triton the sections did not fall you have the solution.
Try sections with/without Triton, evaluate the results and act accordingly.
René J.


________________________________
From: Tyrone Genade <tgenade <@t> gmail.com>
To: histonet <histonet <@t> lists.utsouthwestern.edu> 
Sent: Wednesday, August 15, 2012 8:44 AM
Subject: [Histonet] titron vs DMSO

Hello,

I suspect that 0.1% triton X-100 in my blocking solution may be
causing my frozen (unfixed) sections of brain to come off the slide.

These are APTES slides and the sections were 20 and 40 micron thick.
At first it looked to me that the entire section had come off but
under the microscope I could see a thin layer (of single?) cells still
stuck to the slide. There was also a chance in antibody staining. The
antiserum I am using normally gives very defined intense staining of a
surface antigen but now the staining was punctate. I suspect the
triton is too strong and as well as washing off the tissue it is also
disrupting the membranes and interfering with the staining. I have not
seen this when I was using the same blocking solution without the
triton X-100.

For wholemounts I had used DMSO for permeabilization but also have
NP-40 available.

Does anyone have an opinion on the matter?
-- 
Tyrone Genade
Dept. Human Biology
University of Cape Town

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