Hi, I dont think glutaraldehyde is good for many antigens...though the brain does become firmer...
2009-03-06 TF 发件人: Geoff McAuliffe 发送时间: 2009-03-06 23:37:34 收件人: shymaa shawadfy 抄送: histonet 主题: Re: [Histonet] postnatal brain sections using vibratome 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 祄 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
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