Re: immuno/floating tissue
|From:||Amos Brooks <email@example.com> (by way of histonet)|
Using sailinized or charged slides is a great start. Using Halt or
another adhesive in conjunction with these slides is counter productive.
There has been some discussion on this subject in the past, (you can find
these in the archives). Briefly the reason is the charged surface of the
slide is covered with a neutral layer of adhesive.
Plenty of drying time is also crucial. If there is any residual water on
the slide it will try to come off at some point during the procedure, and
along with it will come the section covering the water / slide.
PS: Sorry this response is so late, I took a long weekend. :-)
----- Original Message -----
From: "Vicki Kalscheur" <firstname.lastname@example.org>
Sent: Friday, July 20, 2001 9:55 AM
Subject: immuno/floating tissue
> I am a graduate student working on my MS in the Comparative
> Research Lab at the UW-Madison.
> My project involves examining chondrocyte viability in bovine patella
> samples over time. Part of my project is examining the articular
> cartilage for apoptotic chondrocytes. I will be using Anti-Caspace
> Anti-PARP, and Tunel assays to do this.
> I am, however, having a problem regarding this procedure. It seems
> that my samples will not adhere to the slides during the assays.
> After about 4 washes (only 1/4 of the way through each assay) the
> sections fall off the slides and float in the solution.
> I have tried Aminoalkylsilane coated slides, and a product called
> to help adhere the sections but have had no luck. I am currently
> using EDTA to decalcify the sections and frozen sectioning. I have
> considered using paraffin embedded samples to secure my samples more
> effectively but am afraid of the damage to the chondrocytes during
> heating. We also plan on trying some free-floating analysis.
> I would greatly appreciate any suggestions anyone could provide. My
> email is email@example.com. My fax is 608-263-7930 (please
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