I have bones of mice that are PFA fixated, cryoembedded and cut into slices with the Kawamoto method. Now I am trying to stain intracellular markers on these sections. I have tried different permeabilization methods and durations (e.g. with Triton-X100) and it works in soft tissues however, I do not get specific signals in bone marrow. Does anyone have experience with that and could help me out?
Thanks in advance,
Radost
Hello, for the expression of proteins by the immunohistochemistry technique, I recommend that you decalcify the bone marrow sections, fix them in 3.7 to 4% formaldehyde in phosphate buffer and then include them in paraffin and first perform an H&E and then the immunohistochemistry .
Dear Radost,
Yes, suggestion by Francisco sounds good. I have heard your method also of plastic sections of bones but I do not have experience of immunocytochemistry on plastic sections. You may e-mail to
Matthew Hilton
Dr. Hilton has experience in plastic sections.
As far as decalcification method is concerned, I have used and two reprints are enclosed where bone and cartilage have been stained after decalcification for many antigens and also for special staining.
Good luck, Dr Hilton will resolve your issue