Even 30um is too thick for the routine IHC on glass. You will never penetrate the whole thickness for staining.
Colleen Forster HT(ASCP)QIHC On Fri, Aug 4, 2023 at 2:05 PM Alonso Martínez Canabal < acana...@ciencias.unam.mx> wrote: > Hi, thank you for your reply. > I cannot do my usual free floating staining because this tissue is > very brittle. So, I was thinking to do the staining in slide at 30 microns? > Thank you > > El vie, 4 ago 2023 a las 12:29, Colleen Forster (<cfors...@umn.edu>) > escribió: > >> 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 >> >> >> >> >> >> >> > > -- > Dr. Alonso Martínez Canabal PhD > Profesor Asociado "C" > Departamento de Biología Celular, Facultad de Ciencias, UNAM > Investigador Nacional "I" > 56224833 > > -- 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