Summary
A technique is described which permits the direct, infrared (IR) spectrophotometric observation of peripheral leukocytes by utilizing multiple internal reflectance (MIR) spectroscopy on zinc selenide prisms. The IR spectra of nitro-blue tetrazolium (NBT) and its derivative, nitro-blue diformazan (NBF), are described and examined and these spectra are compared with those of the intracellular NBT and NBF yielding several peaks which may prove to be analytically useful. Additionally, the presence of unreduced NBT in leukocytes is demonstrated, which microscopic and chemical “NBT tests” have not previously done.
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Baehner, R. L., Nathan, D. G.: Leukocyte oxidase: defective activity in chronic granulomatous disease. Science 155, 835–836 (1967)
Baehner, R. L., Nathan, D. G.: Quantitative nitro-blue tetrazolium test in chronic granulomatous disease. New Engl. J. Med. 278, 971–976 (1968)
De La Vega, G., Freyre-Horta, R., Benitez-Bibriesca, L.: Plasma factor affecting the NBT reducing capacity of neutrophils. New Engl. J. Med. 289, 271–272 (1973)
Gilby, A. C., Cassels, J., Wilks, P. A.: Internal reflectance spectroscopy III. Micro sampling. Appl. Spectrosc. 24, 539–542 (1970)
Humbert, J. R., Marks, M. I., Hathaway, W. E., Thorsen, C. H.: The histochemical nitro-blue tetrazolium reduction test in the differential diagnosis of acute infections. Pediatrics 48, 259–267 (1971)
John, S., Berger, N., Bonner, M. J., Schultz, J.: Localization of lactic dehydrogenase isozymes in lysosomal fraction of the neutrophil of normal human blood. nature (Lond.) 215, 1483–1485 (1967)
Mattson, J. D., Mark, H. B., Weber, W. J., Identification of surface functional groups in active carbon by infrared internal reflection spectrophotometric (sic). Analyt. Chem. 41, 355–358 (1969)
Ochs, H. D., Igo, R. P.: The NBT slide test: a simple screening method for detecting chronic granulomatous disease and female carriers. J. Pediat. 83, 77–82 (1973)
Park, B. H., Fikrig, S. M., Smithwick, E. M.: Infection and nitro-blue tetrazolium reduction by neutrophils: a diagnostic aid. Lancet 1968 II, 532–534
Silverstein, R. M., Bassler, G. C.: Spectrometric identification of organic compounds, vol. 2, p. 68. New York: John Wiley & Sons 1967
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)
Taclob, L. T., Patel, S. R., De Bari, V. A., Needle, M. A.: Decrease in stimulated phagocyte nitro-blue tetrazolium reduction with prolonged hemodialysis. Abstract. Amer. Soc. Neph. p. 91 (1974)
Windhorst, D. H., Holmes, B., Good, R. A.: A newly defined X-linked trait in man with demonstration of the Lyon effect in carrier females. Lancet 1967 I, 737–739
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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). https://doi.org/10.1007/BF00508055
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DOI: https://doi.org/10.1007/BF00508055