New flow cytometry-based method for the assessment of the antibacterial effect of immune cells and subcellular particles

Ákos M. Lorincz, Viktória Szeifert, Balázs Bartos, Erzsébet Ligeti

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Techniques currently used for assessment of bacterial count or growth are time-consuming, offer low throughput, or they are complicated or expensive. The aim of the present work was to elaborate a new method that is able to detect the antibacterial effect of cells, subcellular particles, and soluble compounds in a fast, cost, and labor effective way. Our proposed technique is based on flow cytometry (FC) optimized for detection of small particles and on fluorescently labeled bacteria. It allows direct determination of the bacterial count in 3 hours. The effect of various human phagocytes and extracellular vesicles on gram-positive and gram-negative bacteria is investigated in parallel with the new, FC-based method, with colony counting and with our previous, OD-based method. Comparing the killing effect of wild type and NADPH oxidase-deficient murine neutrophils presents an example of detection of a clinically important deficiency. Strong correlation was obtained between the results of the different techniques, but the reproducibility of the FC-based test was superior to the OD-based test. The major advantages of the new technique are: rapidity, low cost, high throughput, and simplicity.

Original languageEnglish
JournalJournal of Leukocyte Biology
DOIs
Publication statusAccepted/In press - Jan 1 2018

Keywords

  • Bacterial killing
  • Extacellular vesicles
  • Fluorescent staining
  • Immunodeficiency
  • Phagocytes

ASJC Scopus subject areas

  • Immunology
  • Cell Biology

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