I made TRAP staining solution with Naphthol AS-MX(1mg/ml) and Fast red salt (1mg/ml) in DMF/sodium acetate buffer pH 5.0.
but it only work in vitro osteoclast sample, not in vivo tissue sample (femur and cranial bone)
Is there any difference between in vitro and in vivo histology TRAP staining?
my procedure
in vitro : osteoclast culture with bone marrow macrophage (BMM) -> fixation with PFA 4%-> staining with TRAP solution
in vitro : in vivo sample fixation in formaldehyde 10%, decalcification and paraffin embedding -> tissue section -> deparaffinize & hydration -> trap solution apply
p.s.
I think it's difficult to think there is no osteoclast in tissue sample...
is phosphatase activity disappeared at some kind of environment?
I attached in vitro image.
I solved.
Use neutral EDTA solution for the decalcification process when you make histological samples. (acidic decal solution not worked with TRAP)
Thank you for your reply. I also got the same issue as you described.
How did you filter the solution? Eventhough I filtered the solution, I found junks on surface. The cell surface is not clear even after rinsing with DW. Do you have any solution to that problem? I saw your photo is very clear. Could you give any suggestion about my problem? Thanks in advance.
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