The salts which are used for the peroxidase stain of blood cells have been classified into four sorts, of which Salt 1 and 2 are useful, while Salt 3 and 4 are entirely useless for the stain. The available Methods, devised until recently by many investigators, apply the Salt 1 or 2 for the color change of the peroxidase granules. We have also made it clear that the method of showing blue granules such as Sat o's does not differ essentially from that of showing brown granules such as Graham's, because the reactions of the two methods are linked with each other ; the method of showing blue granules in the myelogenous blood cells indicates brown granules at first, which are gradually changed into blue according to the staining time. Of course, both the brown and illue granules can never be seen without benzidine, and the blue granules originate from the brown. Judging from this, it is suggested to us that so-called peroxidase stain of blood cells may be due to the affinity between peroxidase granules and benzidine, and that between oxidized benzidine and salts. The mechanism of this reaction is to be explained in further reports, though it is generally believed that the peroxidase of blood cells is the ferment which resolves hydrogen peroxide as the source of oxygen for the rapid oxidation of benzidine. We are indebted to Professor Osgood of Oregon University and Professor Hayaka wa of our School for their much valuable advice, and wish to express sincere thanks to them. This paper was read before the 55th annual meeting of the Japanese Anatomical Society, September,1950, and the study was carried out with the grant in aid for the foundamerital scientific research.

View full abstract

We have devised a simple peroxidase staining method using nickelammonium sulfate, and named this NAS-Benzidine reaction. By means of this we can stain the peroxidase positive granules at a time and more rapidly than the existing methods which use separate solutions to be mixed freshly each time. The NAS-Benzidine solution including benzidine, hydrogen peroxide and nickel-ammonium sulfate (NAS), keeps for six months without significant loss of potency. An additional advantage of this stain is the fact that it may be used not only for dry smears but also for the staining of sections and fresh blood. This paper has already been read before the 56th annual Japanese Anatomical Society, April,1951, and the study was carried out with the grant in aid for foundamental scientific research.

View full abstract