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This study was carried out in order to establish a simple and stable method to demonstrate acid phosphatase activity of leucocyte on blood smear. Settling a tentative method according to Pearse's azo dye method for tissue section, each experiment was carried out to choose the most suitable condition at each step of the procedure. It was made clear that the following method was a simple and reliable one. 1) Prepare blood or bone marrow smears. 2) Dry the smears. 3) Fix for 5 minutes in neutral formalin vapor at room temperature below 20°C with-in 60 minutes after the smear preparation. 4) Expose the smears to air for 1 hour to remove formalin. 5) Prepare the incubating solution as follows. Fifty mg of α-naphthylphosphate, 50mg of Fast-Garnet-GBC salt, 50ml of acetate buffer (pH 5.0, 0.1M) are mixed, shaken well, and filtrated. 6) Incubate in it for 90 minutes at 37°C. 7) Wash well in distilled water. 8) Counterstain nuclei with Meyer's hematoxylin for 30 minutes. 9) Wash for 5 minutes in tap-water. 10) Mount in 40% PVP and microscopy.

Azo Dye Method for Demonstration of Leucocyte Acid Phosphatase Activity on Blood Smears