Your description gives the impression of damage caused by slow
freezing = without cryoprotection. Ice crystals form in the
tissue, leaving beh= ind holes. Prevention: adequate fixation in
buffered formaldehyde,= followed by cryoprotection (usually soaking
in 30% aqueous sucrose fo= r a day or two; until the brain
sinks). Even with cryoprotection, = the specimen should be frozen
as quickly as possible. There have been = many Histonet postings on
this subject over the years. They can be fou= nd at www.histos earch.com.
John
Kiernan
Anat= omy, UWO
London, Canada.
=
= =
----- Or= iginal Message -----
From:
"Leskovjan, Andreana" < leskovjan@bnl.gov>
Date: Tuesday, April 22,
2008= 12:16
Subject: [Histonet] Thioflavin S
staining
T= o:
histonet@lists.utsouthwestern.edu
> Hi,
>
>
>
&= gt; I need to stain 30 um frozen, unfixed mouse brain
sections with<= BR>> Thioflavin S. The sections
are mounted onto a = very thin
> plastic
film
> called Ultralene= . I know it's not
the best but necessary
> = for our
>
measurements. I have been using a ve= ry simple procedure
so
> far with
> minimal= chemicals to
avoid interference with the
> measurements: 50%
> ethanol for 5 min; 0.0125%
Thiofl= avin S for 3 min; wash in 50%
>
ethanol,then nanopu= re water. The staining works
however
> the ti= ssue is almost
>
destroyed. There are holes,= tears, and folds
everywhere,
> which makes
&= gt; the
plaques difficult to see and measure. I've tried= many
> combinationsof the above and this is the best I c ould get.
> Does anyone have
a
>= suggestion on how to prevent this and get better
looking
>= sections?
>
> >
>
Thanks,
>
> = Andreana
Leskovjan
>
> _______ _______________________ 5F= ________________
>
Histone= t mailing list
>
Histonet@lists.utsouthwestern.edu<= BR>>
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|