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Limitations on the use of dihydrorhodamine 123 for flow cytometric analysis of the neutrophil respiratory burst

van Pelt, L. J., van Zwieten, R., Weening, R. S., Roos, D., Verhoeven, A. J. & Bolscher, B. G., 1996, In : Journal of Immunological Methods. 191, 2, p. 187-96 10 p.

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  • Limitations on the use of dihydrorhodamine 123 for flow cytometric analysis of the neutrophil respiratory burst

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DOI

  • L J van Pelt
  • R van Zwieten
  • R S Weening
  • D Roos
  • A J Verhoeven
  • B G Bolscher

Intracellular oxidation of dihydrorhodamine 123 (DHR) to the fluorescent compound rhodamine 123 (Rho123) was used to detect the production of oxygen metabolites in activated neutrophils. Total leukocyte preparations can be used in this assay, which is a great advantage when priming of the respiratory burst is studied. We have defined the conditions that should be taken into account when priming is studied with this assay. We found that neither the extent nor the kinetics of DHR oxidation match those of NADPH oxidase activity. In addition, DHR oxidation is influenced by the absolute and relative number of neutrophils in the leukocyte suspension, by the DHR concentration and by myeloperoxidase availability. The results presented in this study emphasize the need for carefully designed experiments when DHR is used to study the respiratory burst in neutrophils.

Original languageEnglish
Pages (from-to)187-96
Number of pages10
JournalJournal of Immunological Methods
Volume191
Issue number2
Publication statusPublished - 1996
Externally publishedYes

    Keywords

  • Azides, Catalase, Cell Separation, Female, Flow Cytometry, Humans, Indicators and Reagents, Male, Neutrophils/chemistry, Peroxidase/pharmacology, Respiratory Burst/drug effects, Rhodamines/metabolism

ID: 93120699