[Histonet] problem with brain sections falling off subbed slides
jkiernan <@t> uwo.ca
Wed Nov 2 22:19:22 CDT 2011
Gelatin alone is not an adhesive because it dissolves in water. Chrome alum-gelatin is an excellent adhesive for sections and costs almost nothing to make. Subbed slides are one coated with chrome gelatin, dried and stored. The following web link shows a 1999 publication giving the rationale and methods for a variety of section adhesives.
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On 02/11/11, Douglas M Burns <dmburns9 <@t> gmail.com> wrote:
> I am new to working with brain, and I am having a bad problem with
> both thin and thick rat brain sections sliding off of subbed slides.
> We tried commercial treated slides first, but then moved on to
> subbing them. The subbing was done with gelatin according to a printed
> protocol, and I can see the film on the dried slide. When kept horizontal
> they seem okay, but as soon as they are put into a slide holder for washing
> or anything, they begin to slide off or even move around. After a short
> while, many/most of mysteriously vanished (into the wash solution).
> Our slices are sliced by cryostat from a rat brain that has been
> fixed with 4% paraformaldehyde and 30% sucrose. The slice comes right off
> the cryostat, and then I m ove it onto a warm slide. (Perhaps they are not
> warm enough?? I have seen others blow on them to warm them.) Slides
> destined for Cresyl violet staining were dried for 3 days at room
> temperature before being moved.
> It is the Cresyl violet protocol that causes the most trouble, but I
> am seeing some problems with any type of large volume wash. At this point,
> I don't have enough experience with this specific tissue and our protocols
> to be able to tell for sure what is happening. Thus, I am entirely open to
> advice - any advice that might help with the problem.
> Thank you ------------- Doug Burns, Kansas City
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