slide coating/in situ query
<< Previous Message | Next Message >>
From: | "Marshall, Sharon, Mrs" <marshall@anat.uct.ac.za> |
To: | histonet@pathology.swmed.edu |
Reply-To: | |
Date: | Mon, 20 Sep 1999 16:34:06 SAST-2 |
Content-Type: | text/plain; charset=US-ASCII |
Dear histonetters,
Would like some advice on the following. We wish to coat some slides
with APTES before doing in situ hybridisation. I would prefer to use
extran as an initial wash as opposed to chromic acid as I feel more
comfortable working with a detergent than an acid. I am using the
extran at a dilution of 10%. Is this too high or could I get away
with a lower dilution? Apparently the acid would destroy any
protein present. Would the detergent do this as well? I plan to leave
the slides in extran overnight. The method I have also recommends a
wash after the extran step at 60 degrees for 2hrs in tap water. Could
anybody explain or has a gremlin crept in when the method was passed
along. I presume that washing in tap water for 2hrs at R.T. would
be just as sufficient? The next step is a drying procedure at 160
degrees for 2hrs. I guess any drying method is suitable? Once the
slides haved cooled we will then coat them, (2% Aptes in
acetone)5secs., rinse them in acetone and sterile distilled water and
dry them. We also then DEPC treat and bake the slides before using
the slides for in situ.
Thanks
Sharon Marshall
e-mail: marshall@anat.uct.ac.za
<< Previous Message | Next Message >>