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Quantification of heterotypic granule fusion in human neutrophils by imaging flow cytometry.

Journal article
Authors Halla Björnsdottir
Amanda Welin
Claes Dahlgren
Anna Karlsson
Johan Bylund
Published in Data in brief
Volume 6
Pages 386-93
ISSN 2352-3409
Publication year 2016
Published at Institute of Odontology
Institute of Medicine, Department of Rheumatology and Inflammation Research
Institute of Odontology, Section 3
Pages 386-93
Language en
Subject categories Other Medical Sciences


Human neutrophils are filled with intracellular storage organelles, called granules and secretory vesicles, which differ in their content of soluble matrix proteins and membrane-bound molecules. To date, at least four distinct granule/vesicle subsets have been identified. These organelles may secrete their content extracellularly following mobilization to and fusion with the plasma membrane, but some of them may also fuse with internal membrane-enclosed organelles, typically a plasma membrane-derived phagosome. There are also instances where different granules appear to fuse with one another, a process that would enable mixing of their matrix and membrane components. Such granule fusion enables e.g., myeloperoxidase-processing of intragranular oxygen radicals, a key event in the formation of neutrophil extracellular traps (Björnsdottir et al., 2015) [1]. Described herein are data that show the quantification of such heterotypic granule-granule fusion by the use of imaging flow cytometry, a technique that combines flow cytometry with microscopy. The analysis described is based on immunofluorescent staining of established granule markers (lactoferrin and/or NGAL for one granule subset; the specific granules, and CD63 for another granule subset, the azurophil granules) and calculation of a colocalization score for resting and PMA-stimulated neutrophils.

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