TRAP Enzyme Histochemistry for Osteoclast Detection on Bone Sections now Established with TissueSurgeon
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The detection of osteoclasts in bone section is not easy, as the cells do not appear in large numbers. They are multinucleated (up to 10 nuclei) and can be found on the bone surface in resorption bays, also called Howships lacunae. A very established way of osteoclast detection is TRAP staining (Tartrate Resistant Acid Phosphatase) which actually detects acid phosphatase enzyme vesicles inside the ostecoclasts. This method is used on either decalcified paraffin sections or ground sections.
For the first time TRAP staining method was proofed on non-decalcified thin sections performed with the Lasermicrotome TissueSurgeon at 10µm thickness. In comparison to decalcified paraffin sections, also hard tissue information is still available for analysis; compared to ground sections, laser sections are much thinner and therefore accessibility to microscopy is improved. In ground sections osteoclasts may be covered by other cell layer.
Preparation of thin sections is much faster and easier to perform with the laser compared to ground sectioning or paraffin sectioning. The establishment of TRAP enzyme histochemistry on thin sections cut with the lasermicotome TissueSurgeon demonstrates that the laser cutting does not destroy the enzyme function and therefore is suitable for enzyme histochemical analysis.