, Volume 56, Issue 2, pp 155–163 | Cite as

Mechanism for transport of nitro-blue tetrazolium into viable and non-viable leukocytes

  • V. A. DeBari
  • M. A. Needle
  • A. Prado


The permeability of the membrane of phagocytic leukocytes (WBC) to the non-complexed nitro-blue tetrazolium (NBT) cation has not been well explored. In this study both complexed and non-complexed (or free) NBT have been incubated with viable cells. The entry of NBT into the WBC was determined by both colorimetric and infrared analyses. Very little free NBT enters the viable WBC compared with NBT complexed with serum factors. NBT entry was also studied in non-viable WBC by use of the infrared system. The entry of NBT into the non-viable cells was independent of serum complexing factors. Much larger quantities of both free and complexed NBT enter non-viable cells than enter viable WBC. It is concluded that the membrane of the viable WBC is quite impermeable to the free NBT cation. Ultrastructural studies suggest that the gaps in the plasma membrane of non-viable cells could allow the passage of relatively large amounts of both free and complexed NBT.


Public Health Plasma Membrane Viable Cell Tetrazolium Ultrastructural Study 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Altman, F.P.: Quantitative dehydrogenase histochemistry with special reference to the pentose shunt dehydrogenases. Prog. Histochem. Cytochem. 4, 225–273 (1972)Google Scholar
  2. Altman, F.P.: Studies on the reduction of tetrazolium salts III. The products of chemical and enzymic reduction. Histochemistry 38, 155–171 (1974)Google Scholar
  3. Baehner, R.L., Nathan, D.G.: Leukocyte oxidase: defective activity in chronic granulomatous disease. Science 155, 835–836 (1967)Google Scholar
  4. Baehner, R.L., Nathan, D.G.: Quantitative nitro-blue tetrazolium test in chronic granulomatous disease. New Eng. J. Med. 278, 971–976 (1968)Google Scholar
  5. Bainton, D.F., Farquhar, M.G.: Differences in enzyme content of azurophil and specific granules of polymorphonuclear leukocytes. II. Cytochemistry and electron microscopy of bone marrow cells. J. Cell Biol. 39, 299–317 (1968)Google Scholar
  6. Czarnetzki, B.M., Cowan, D.H., Belcher, R.W.: The effects of polyanions on NBT reduction, hexose monophosphate shunt activity, and ultrastructure of polymorphonuclear leukocytes. Amer. J. Clin. Pathol. 64, 34–40 (1975)Google Scholar
  7. DeBari, V.A., Coste, J.F., Needle, M.A.: Direct spectrophotometric observation of intracellular nitro-blue tetrazolium and its formazan by multiple internal reflectance infrared spectroscopy. Histochemistry 45, 83–88 (1975)Google Scholar
  8. DeBari, V.A., Needle, M.A.: The adhesion of blood phagocytes to zinc selenide studied by reflectance-infrared spectroscopy. J. Recticuloendothel. Soc. 22, 121–128 (1977)Google Scholar
  9. Handin, R.I., Stossel, T.P.: Phagocytosis of antibody-coated platelets by human granulocytes. New Eng. J. Med. 290, 989–993 (1974)Google Scholar
  10. Matula, G., Paterson, P.Y.: Spontaneous in vitro reduction of nitroblue tetrazolium by neutrophils of adult patients with bacterial infection. New Engl. J. Med. 285, 311–317 (1971)Google Scholar
  11. Park, B.H., Fikrig, S.M. and Smithwick, E.M.: Infection and nitro-blue tetrazolium reduction by neutrophils. A diagnostic aid. Lancet 1968/II, 532–534Google Scholar
  12. Pearson, B., Defendi, V.: Histochemical demonstration of succinic dehydrogenase in thin tissue sections by means of 2-(p-iodopheny)-3(p-nitrophenyl)-5-phenyl tetrazolium chloride under aerobic conditions. J. Histochem. Cytochem. 2, 248–257 (1954)Google Scholar
  13. Rothwell, D.J., Doumas, B.T.: The effect of heparin and EDTA on the NBT test. J. Lab. Clin. Med. 85, 950–956 (1975)Google Scholar
  14. Rutenberg, A.M., Gofstein, R. and Seligman, A.M.: Preparation of a new tetrazolium salt which yields a blue pigment on reduction and its use in the demonstration of enzymes in normal and neoplastic tissues. Cancer Res. 10, 113–121 (1950)Google Scholar
  15. Segal, A.W., Levi, A.J.: The mechanism of the entry of dye into neutrophils in the nitro-blue tetrazolium (NBT) test. Clin. Sci. Mol. Med. 45, 817–826 (1973)Google Scholar
  16. Segal, A.W., Levi, A.J.: Factors influencing the entry of dye into neutrophil leukocytes in the nitro-blue tetrazolium test. Clin. Sci. Mol. Med. 48, 201–212 (1975)Google Scholar
  17. Stossel, T.P.: Evaluation of opsonic and leukocyte function with a spectrophotometric test in patients with infection and with phagocytic disorders. Blood 42, 121–130 (1973)Google Scholar
  18. Taclob, L.T., Patel, S.R., DeBari, V.A., Needle, M.A.: Decrease in stimulated phagocyte nitro-blue tetrazolium reduction with prolonged hemodialysis. (Abstract) Amer. Soc. Neph. 7, 91 (1974)Google Scholar

Copyright information

© Springer-Verlag 1978

Authors and Affiliations

  • V. A. DeBari
    • 1
    • 2
  • M. A. Needle
    • 1
    • 2
  • A. Prado
    • 3
  1. 1.Renal Division and Department of MedicineSt. Joseph's Hospital and Medical CenterPatersonUSA
  2. 2.College of Medicine and DentistryNew Jersey Medical SchoolNewarkUSA
  3. 3.Department of PathologyBarnert Memorial Hospital CenterPatersonUSA

Personalised recommendations