If you put a little glutaraldehyde (0.25%) in the fixaitve you perfuse
with but not in the post fix solution the brain will be noticably firmer
and your antigen may survive.
Good luck!
Geoff
shymaa shawadfy 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
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcaul...@umdnj.edu
**********************************************
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
