Currently doing Cryotomy on quickly frozen Golgi-Cox impregnated mouse brains at 100-120 microns. Cryoprotecting with 30% sucrose. Cutting at -15C. Tried fresh tissue and brief NBF perfusion(5 min). If I perfuse with NBF for 15 min, or immersion fix overnight, it seems to destroy spines and create artifact- possibly glial cells. Cutting much easier though!
Might Histogel help? Bob Nienhuis UCLA / VA Medical Center On Tue, Jan 13, 2009 at 2:10 PM, gayle callis <gayle.cal...@bresnan.net>wrote: > Bob, > > So which type of sectioning are you doing now? Cryotomy or vibratome? And > how thick do you want the sections to be? If frozen sections on snap > frozen > mouse brain, simply turning the temperature up to -16C or so will make > sectioning much easier and get rid of the shredding, friable sections. > Also, are you fixing then sucrose cryoprotecting after NBF or 4% > paraformaldehyde fixation of the brain (can be either perfused or immersion > fixation)? > > Gayle Callis > HTL(ASCP)HT,MT > Bozeman MT > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet