Re: free floating sections

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From:"Barry Rittman" <> (by way of Marvin Hanna)
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I have stained lots of free floating sections and the method recommended by
works well. A problem I found was that if you are not careful when you change
fluids the section may tend to stick to the plastic.
We have traditionally used either welled slides or cavity dishes (with 6-12
each). Both made either from glass or from porcelain.
These both have the advantage of having concave wells which use less
solution. The
concave wells allow transfer of sections (because sections have to be
mounted). Glass "hockey sticks" can be introduced from the side as the edges of
each well are shallow and sloping. While this can be done with the 24 well
as Geoff describes, I found this to be more difficult as the side of the tissue
culture wells is straight. This often necessitates the hockey stick being
held at
an uncomfortable angle especially with large sections.
The big advantage of the 24 well tissue culture dishes is being able to
throw them
away after use. Welled slides and cavity dishes need to be cleaned.

Geoff McAuliffe wrote:

> Cynthia Favara wrote:
> > All,
> >         I have been asked to stain free floating sections and am enlisting
> > my greatest resource. If possible we would like to use frozen tissue and
> > thick sections. The premise is that we can stain without the tissue ever
> > drying. Advice from anyone with experience would be greatly appreciated.
> > I'll take any ideas and promise not to be offended by sarcasm etc [polly
> > would definitely freeze here today]
> > TIA,
>     I stain 20 micron frozen sections of fixed mouse (or rat) brain in 24
> tissue culture plates. I use 300 microliters per well for 2-3 sections (more
> sections if they are small) and do the incubations on a shaker table. If you
> put too much solution in the wells you will not get proper agitation. I
> solutions by pipetting, no need to move sections. Working against a black
> background makes it much easier to see the sections when changing solutions!
> Geoff
> --
> **********************************************
> Geoff McAuliffe, Ph.D.
> Neuroscience and Cell Biology
> Robert Wood Johnson Medical School
> 675 Hoes Lane, Piscataway, NJ 08854
> voice: (732)-235-4583; fax: -4029
> **********************************************

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