Alonso, For sections that thick you would collect them as you always have, stain them as floating sections and then mount onto a glass slide. The routine standard methods for IHC will never penetrate a 50um thick section.
Colleen Forster HT(ASCP)QIHC On Fri, Aug 4, 2023 at 1:17 PM Alonso Martínez Canabal via Histonet < histonet@lists.utsouthwestern.edu> wrote: > Good afternoon Histoneters, > I have been doing IHC and IF during decades in rodent brain tissue, > always in free floating. Now, I would like to do that sections placed over > slides and perform the whole procedure over the slide. I am interested in > using thick sections, usually I used 50micrometers, what is the maximum > thickness that I could use in slide to guarantee the correct penetration of > the antibodies and the ABC complex. Before the IHC I always use graded > alcohols and xilene, and antigen retrieval with citrate buffer. > Thank you! > > -- > Dr. Alonso Martínez Canabal PhD > Profesor Asociado "C" > Departamento de Biología Celular, Facultad de Ciencias, UNAM > Investigador Nacional "I" > 56224833 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Colleen Forster HT(ASCP)QIHC BLS Histology and IHC Laboratory Jackson Hall, Room 2-155 321 Church St. SE Minneapolis, MN 55455 612-626-1930 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet