Cold acetone fixation and methacrylate embedding. A suitable method for routine processing of bone marrow biopsies.

Here we report that acetone fixation at -18 degrees C with subsequent embedding in methyl-/butylmethacrylate is a reliable method for the routine processing of bone marrow biopsies. This method allows good conventional histological visualization of morphological details, which is comparable with other fixation procedures. The essential advantage of this method is that a wide range of monoclonal antibodies and polyclonal antisera can be used for immunohistochemical investigations for diagnostic and scientific purposes. The addition of 5% polyethylene glycol 400 to the acetone minimizes freeze-related artefacts. The immunohistochemical demonstration of a number of antigens is mostly affected by the medium used for slide preparation and to a lesser extent by the concentration of benzoylperoxide used for polymerization. Performing polymerization at 4 degrees C and using N, N-dimethyl-p-toluidine as accelerator allows the concentration of benzoylperoxide to be reduced to 0.2 g% (8.3 mmol). Under these conditions the methacrylate embedding procedure has only minimal effects on the quality of immuno- and enzyme histochemistry. Additionally, the simplified method for removing the polymerization inhibitor from the methacrylate components and the shortened impregnation step are further advantages of the embedding method described here.