Hi, The thicker the section the more likely it will be to fall off. Frozen sections already love to fall off slides. You should cut them a lot thinner. 4 to 10 um should be the thickness, especially for frozens. If you want very thick sections as you describe, you would be better off cutting them and transferring them directly to an 8 well plate with PBS and doing the IHC as a floating section.
Cheers, Amos Brooks > > Message: 1 > Date: Tue, 19 Sep 2023 19:07:04 -0600 > From: Alonso Mart?nez Canabal <acana...@ciencias.unam.mx> > To: Histonet <histonet@lists.utsouthwestern.edu> > Subject: [Histonet] p17 mice brain sections > Message-ID: > <CAG12gaHPkXVjnHO= > wdugsdppx5ytkxxuaipmwctzjcazwf_...@mail.gmail.com> > Content-Type: text/plain; charset="UTF-8" > > Hello, > Have a great afternoon. I have done HAR with citrate buffer (pH 6.0) > with tween, basically all my professional life of some 15 years. Several > publications, using mice brain sections 40-50 microns thickness from > cryostat (30% sucrose). Today I tried to do some p17 brain sections and the > sections did not only fell off, but were completely destroyed. > That never happened to me, I am wonder if anyone can have any idea of > what happened? > > Thank you so much > > -- > Dr. Alonso Mart?nez Canabal PhD > Profesor Asociado "C" > Departamento de Biolog?a Celular, Facultad de Ciencias, UNAM > Investigador Nacional "I" > 56224833 > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet