[Histonet] postnatal brain sections using vibratome

[shymaa shawadfy]

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