HISTOCHEMICAL STUDIES ON THE DISTRIBUTION OF ENZYMES, ESPECIALLY OXIDASES AND PHOSPHATASES IN THE LIVING BODY

Two giant salamanders, Megalobatrachus japonicus were used for materials. The blood was carefully taken from the tail and smear preparations were obtained. The blood smears were fixed one minute with ethanolformol 91 solution prior to staining, washed thoroughly in distilled water, and stained five minutes with either of the following peroxidase reagents freshly prepared each time. 1 Mixture of benzidine 0.3 gr., 3 H2O2 10 drops, and distilled water 100 cc. This mixture should be filtered because of poor solubility of benzidine in water. 2 Mixture of orthophenylenediamine 0.1 gr., 3 H2O2 10 drops, and distilled water 100 cc. The smears were then washed in water, and counterstained with dilute Giemsastain for ten to fifteen minutes. Finally they were washed in water, and exposed to air till dry. The alkaline phosphatase technic modified technic of the leadsulphide method of Takeuchi was also applied for blood staining.