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Release of insulin granules by simultaneous, high‐speed correlative SICM‐FCM
Author(s) -
BEDNARSKA J.,
NOVAK P.,
KORCHEV Y.,
RORSMAN P.,
TARASOV A.I.,
SHEVCHUK ANDREW
Publication year - 2021
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/jmi.12972
Subject(s) - exocytosis , insulin , confocal microscopy , chemistry , confocal , secretory vesicle , vesicle , microbiology and biotechnology , biophysics , fluorescence microscope , fluorescence , secretion , biology , biochemistry , endocrinology , membrane , physics , quantum mechanics , optics
Summary Exocytosis of peptides and steroids stored in a dense core vesicular (DCV) form is the final step of every secretory pathway, indispensable for the function of nervous, endocrine and immune systems. The lack of live imaging techniques capable of direct, label‐free visualisation of DCV release makes many aspects of the exocytotic process inaccessible to investigation. We describe the application of correlative scanning ion conductance and fluorescence confocal microscopy (SICM‐FCM) to study the exocytosis of individual granules of insulin from the top, nonadherent, surface of pancreatic β‐cells. Using SICM‐FCM, we were first to directly follow the topographical changes associated with physiologically induced release of insulin DCVs. This allowed us to report the kinetics of the full fusion of the insulin vesicle as well as the subsequent solubilisation of the released insulin crystal.