Re: [Histonet] Thioflavin S staining

From:John Kiernan

   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
John Kiernan
Anat= omy, UWO
London, Canada.
= = =
----- Or= iginal Message -----
From: "Leskovjan, Andreana" <>
Date: Tuesday, April 22, 2008= 12:16
Subject: [Histonet] Thioflavin S staining
T= o:

> 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
><= BR>> tonet
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