Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
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Imaging Analysis of Insulin Secretion with Two-Photon Microscopy
Noriko Takahashi
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2015 Volume 38 Issue 5 Pages 656-662

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Abstract

High-resolution deep tissue imaging is possible with two-photon excitation microscopy. With the combined application of two-photon imaging and perfusion with a polar fluorescent tracer, we have established a method to detect exocytic events inside secretory tissues. This method displays the spatiotemporal distribution of exocytic sites, dynamics of fusion pores, and modes of exocytosis. In glucose-stimulated pancreatic islets, exocytic events were observed to be synchronized with an increase in cytosolic Ca2+ concentrations. Full fusion of a single secretory granule is the typical mode of exocytosis and compound exocytosis is inhibited. Because two-photon excitation enables simultaneous multicolor imaging due to the broadened excitation spectra, the distributions and conformational changes in fluorescent-labeled molecules can be simultaneously visualized with exocytic events. Therefore, we can analyze the dynamics of the molecules involved in membrane fusion and their association with exocytosis in living tissues.

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© 2015 The Pharmaceutical Society of Japan
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