[Histonet] postnatal brain sections using vibratome

[Merced Leiker]

Yes, definitely make sure to do the 20% sucrose overnight, even try 30%, or 
grades going from 10%-20%-30%, switching to the next higher when the brain 
becomes saturated enough to fall to the bottom of the container.  We did 
this in a previous lab with rat brains and it worked well.

Merced

--On Friday, March 06, 2009 11:52 AM +0900 shymaa shawadfy 
<sshawdfy <@t> med.kobe-u.ac.jp> wrote:

> Dear all
>
> I am trying to use vibratome 50 µm thick sections for immunofluorescence
> using Postnatal day 0 brains. The problem is that brains are very soft and
> are usually destroyed upon handling and the agarose is separated form the
> brain.
>
> My used protocol was: perfusion with 4 % PFA for 3 min, followed by
> several hours to overnight post-fixation. Then embedding brains in 2 %
> low melting agarose and cutting the block on vibratome using low speed.
>
>
>
> I am thinking to add an overnight  20 % sucrose incubation step following
> the post-fixation step. Then embed in agarose and continue the normal
> protocol.  May be sucrose will increase the elasticity of the tissue.
>
>
>
> So what do you think ?
>
>
>
>
>
> Thanks a lot
>
> shymaa
>
>
>
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Merced M Leiker
Research Technician II
354 Biomedical Research Building
School of Medicine and Biomedical Sciences
State University of New York at Buffalo
3435 Main St, Buffalo, NY 14214
Ph: (716) 829-6033
Fx: (716) 829-2725

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