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Simultaneous Flow Cytometric Analysis of Phagocytosis and Oxidative Burst Activity in Equine Leukocytes |
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| Authors: | Flaminio MJBF Rush BR Davis EG Hennessy K Shuman W Wilkerson MJ |
| Institution: | (1) The James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Hungerford Hill Road, Ithaca, New York 14853, USA;(2) Department of Clinical Sciences, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas, USA;(3) Agriculture and Animal Health Division, Bayer Corporation, Merriam, Kansas, USA;(4) Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas, USA |
| Abstract: | This paper describes a method for simultaneously measuring phagocytosis and oxidative burst activity in equine peripheral blood leukocytes by flow cytometry. Opsonized propidium iodide-labelled Staphylococcus aureus (PI-Sa) was used to measure the uptake of bacteria by equine phacocytes and the oxidative burst activity by oxidation of dihydrorhodamine 123. The requirements to achieve optimal activity of phagocytosis and oxidative burst are described. The advantage of the simultaneous technique is that it provides both independent and comparative values for phagocytosis and the oxidative burst, for the detection of impaired mechanisms of microbial destruction. Furthermore, the technique allows evaluation of opsonization activity in this context. |
| Keywords: | flow cytometry horse leukocytes phagocytosis propidium iodide oxidative burst rhodamine |
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