Hi Peter, We use 20-25% sucrose on our fresh specimens for immunofluorescence (IF). We only deal with human skin samples so I will not comment on feline skin. I sit the specimen in three changes of sucrose for 10 minutes each. Before embedding in the cryostat, I gently blot it on a tissue. My understanding is that the sucrose does give support like a matrix, and it has saved dried specimens for me in the past.
I hope this helps. Julie Bilkey. Sydney Skin Pathology. On Fri, Oct 7, 2011 at 7:54 AM, Peter Bodig <pbo...@histoprep.com> wrote: > I have some feline skin samples for frozen sections that were sent to us > because our client was not able to section them themselves. Unfortunately we > are having the same problems trying to make good sections ourselves. The > samples seem dry and unsupported when we try to cut them. The tissue breaks > into small pieces and falls out or the OCT when we try to cut it. We have > tried fixation with 10% NBF for various times up to 24 hrs prior to doing > frozen sections to give the tissue better support, which helps, but not > enough, and then the tissue falls off the coated slides when we try to stain > them. Sectioning at 8µ helps as well (that is as thick as out cryostat will > go). > > > > Does anyone have suggestions on how to get good quality frozen sections > from feline skin? > > > > We have come up with some questions as well: > > > > 1. Is there a way to support the tissue by infiltrating it with OCT or > other similar media to improve our sections? > 2. I have read about using 30% sucrose solution for cryo protection, > does that aide in making good quality sections or just reduce freezing > artifact? > 3. Does tissue need to be fixed prior to using cryo protection? > > > > Our client will be using the sections for Senescence β-Galactosidase > Staining. > > > > Thank you for any help you can provide, > > > > > > Péter Bodig > > Laboratory Coordinator > > Colorado Histo-Prep > > E-mail: pbo...@histoprep.com > > > > _______________________________________________ > 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