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Monitoring real-time hormone release kinetics: Via high-content 3-D imaging of compensatory endocytosis

Journal article
Authors A. I. Tarasov
J. Galvanovskis
O. Rorsman
A. Hamilton
E. Vergari
P. R. V. Johnson
F. Reimann
Frances M. Ashcroft
Patrik Rorsman
Published in Lab on a Chip
Volume 18
Issue 18
Pages 2838-2848
Publication year 2018
Published at Institute of Neuroscience and Physiology, Department of Physiology
Pages 2838-2848
Language en
Links dx.doi.org/10.1039/c8lc00417j
Subject categories Neurosciences

Abstract

High-content real-time imaging of hormone secretion in tissues or cell populations is a challenging task, which is unlikely to be resolved directly, despite immense translational value. We approach this problem indirectly, using compensatory endocytosis, a process that closely follows exocytosis in the cell, as a surrogate read-out for secretion. The tissue is immobilized in an open-air perifusion chamber and imaged using a two-photon microscope. A fluorescent polar tracer, perifused through the experimental circuit, gets trapped into the cells via endocytosis, and is quantified using a feature-detection algorithm. The signal of the tracer that accumulates into the endocytotic system reliably reflects stimulated exocytosis, which is demonstrated via co-imaging of the latter using existing reporters. A high signal-to-noise ratio and compatibility with multisensor imaging affords the real-time quantification of the secretion at the tissue/population level, whereas the cumulative nature of the signal allows imprinting of the “secretory history” within each cell. The technology works for several cell types, reflects disease progression and can be used for human tissue.

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