Release of insulin granules by simultaneous, high-speed correlative SICM-FCM

J Bednarska, P Novak, Yuri E. Korchev, Patrick Rorsman, Andrei Tarasov, Andrew Shevchuk

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Abstract

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, non‐adherent, 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.
Original languageEnglish
Number of pages9
JournalJournal of Microscopy
Early online date22 Oct 2020
DOIs
Publication statusE-pub ahead of print - 22 Oct 2020

Keywords

  • dense core vesicle
  • fusion pore
  • hormone
  • secretion

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