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. 1995 Dec 1;36(4):553–562. doi: 10.1186/BF03547669

Flow-cytometric Studies of the Phagocytic Capacities of Equine Neutrophils

Flödescytometriska studier av fagocytosförmågan hos ekvina neutrofiler

A Johannisson 116,✉, G Gröndahl 216, S Demmers 216, M Jensen-Waern 216
PMCID: PMC8095419  PMID: 8669382

Abstract

Methodological aspects of flow-cytometric evaluation of the phagocytic properties of equine neutrophils were elucidated. The kinetics of attachment and ingestion were studied, and the phagocytic process was more rapidly completed when serum-opsonized yeast cells were used than with use of IgG-opsonized yeast cells. Trypan blue was successfully used to quench fluorescence of non-ingested yeast cells. There were only minor differences in the kinetics of phagocytosis between quenched and un-quenched samples, indicating that attachment is rapidly followed by ingestion. Trypan blue quenching caused loss of cells with light scattering properties of granulocytes, although this did not affect the determined frequencies of truly phagocytic neutrophils. Aggregation of yeast cells proved to be a disturbance but not an obstacle to the determination of frequencies of actively phagocytic cells. Flow cytometry is well suited for studies of phagocytosis of yeast cells by equine neutrophils, and the trypan blue quenching provides a means of eliminating false-positive events due to aggregation of yeast cells. The main advantage of the flow-cytometric method is the possibility of rapid processing of a large number of samples, making the method useful for studies of herds.

Keywords: horses, granulocytes, trypan blue quenching, adhesion, kinetics, ingestion, fluorescence, yeast cells, opsonization

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Acknowledgments

The financial support from the Swedish Horse Race Totalizator Board (ATG) is gratefully acknowledged. The flow cytometer used in this study was financed by a grant from the Knut and Alice Wallenberg foundation.

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