Quantitative Efferocytosis Assays.
Efferocytosis, the phagocytic removal of apoptotic cells, is a dynamic process requiring recruitment of numerous regulatory proteins to forming efferosomes in a tightly regulated manner. Herein we describe microscopy-based methods for the enumeration of efferocytic events and characterization of the spatiotemporal dynamics of signaling molecule recruitment to efferosomes, using genetically encoded probes and immunofluorescent labeling. While these methods are illustrated using macrophages, they are applicable to any efferocytic cell type.
Department of Microbiology and Immunology and the Centre for Human Immunology, The University of Western Ontario, London, ON, N6A 5C1, Canada.
Amanda L
AL
Evans
Department of Microbiology and Immunology and the Centre for Human Immunology, The University of Western Ontario, London, ON, N6A 5C1, Canada.
Jack W D
JW
Blackburn
Department of Microbiology and Immunology and the Centre for Human Immunology, The University of Western Ontario, London, ON, N6A 5C1, Canada.
Charles
C
Yin
Department of Microbiology and Immunology and the Centre for Human Immunology, The University of Western Ontario, London, ON, N6A 5C1, Canada. bheit@uwo.ca.
Bryan
B
Heit
D017209:Apoptosis; D001681:Biological Assay / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006801:Humans; D019169:Jurkat Cells; D008264:Macrophages / Q000166:cytology; D010587:Phagocytosis; D014162:Transfection
Efferocytosis; Immunostaining; Macrophage; Method; Microscopy; Phagocytosis