Blood Journal
Leading the way in experimental and clinical research in hematology

A Histochemical Procedure for Localizing and Evaluating Leukocyte Alkaline Phosphatase Activity in Smears of Blood and Marrow

  1. Laboratories of the Mary Fletcher Hospital and the Department of Pathology, College of Medicine, University of Vermont and State Agricultural College, Burlington, Vermont.


A rapid and simple method is described for demonstrating the presence of alkaline phosphatase activity in cells of blood and bone marrow. Smears were fixed with 10 per cent formalin in methanol at 0 ± 5 C. for 30 seconds and stained by the azo dye procedure, using propanediol buffer at a pH of 9.6.

A "scoring" technic is outlined, which permits the comparison of alkaline phosphatase activities of blood leukocytes of different individuals.

Alkaline phosphatase activity was found only in neutrophilic granulocytes and was localized exclusively in the cytoplasm of these cells. In normal adults, 43 to 95 per cent (mean of 78%) of all neutrophils were unstained. Staining was up to ten times more intense in smears from patients with a variety of pathologic disorders.