Journal
CYTOMETRY PART A
Volume 85, Issue 4, Pages 313-321Publisher
WILEY-BLACKWELL
DOI: 10.1002/cyto.a.22400
Keywords
phagocytosis; bead uptake; time-resolved; flow cytometry; monocyte
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Funding
- Nepean Medical Foundation
- Cure Cancer Australia Foundation
- Leukemia Foundation of Australia
- ARC [FT120100581]
- NHMRC Project [1048082]
- Australian Research Council [FT120100581] Funding Source: Australian Research Council
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Phagocytosis is central to immunity however a rapid and standardized method is much needed for quantitative assessment of the phagocytic process. We describe a real-time flow cytometric method to quantitate the phagocytosis of fluorescent latex beads by human monocytes in serum-free conditions. Effects of buffer composition, temperature, pH, and bead surface on phagocytic rate are described. The innate phagocytic ability of human monocytes from single subjects measured by this method was relatively stable over many months although phagocytosis rate varied as much as two-fold between individuals. Comparable results were obtained with a simplified method using several mL of whole blood which is suitable for routine clinical application. This method also allows two-color flow cytometric measurement of cytosolic calcium levels during the phagocytic uptake of fluorescent beads. (c) 2013 International Society for Advancement of Cytometry
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