Examining the kinetics of Neutrophil phagocytosis by flow cytometry

Two methods for measuring phagocytosis are routinely used, microscopy and flow cytometry. Both techniques measure the capability of phagocytic cell to ingest a fluorescent particle (labeled microorganisms or beads).  There are several advantages to using flow cytometry to measure phagocytosis. Microscopy requires time consuming manual counting of ingested particles, which limits the total number of cells analyzed. Conversely, flow cytometry can analyze thousands of cells per second facilitating statistical analysis while offering multi-parametric measurements for simultaneous evaluation of neutrophil features. Finally, neutrophils are easy to prime and activate by manipulation in the lab, which is a necessary process for microscopic analysis of phagocytosis. Flow cytometry allows the measurement of neutrophil phagocytosis in white blood cells without creating artifacts and therefore may be more representative of phagocytosis in vivo. In this application note, we measured neutrophil phagocytosis on the NovoCyte flow cytometer under various conditions and quantified the effect of a potent actin remodeling inhibitor, cytochalasin D.

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