Imaging Single-Vesicle Exocytosis with Total Internal Reflection Fluorescence Microscopy (TIRFM)

Yingke Xu; Luhong Jin; Derek Toomre

doi:10.1007/978-1-0716-2209-4_12

Imaging Single-Vesicle Exocytosis with Total Internal Reflection Fluorescence Microscopy (TIRFM)

Methods Mol Biol. 2022:2473:157-164. doi: 10.1007/978-1-0716-2209-4_12.

Authors

Yingke Xu¹, Luhong Jin², Derek Toomre³

Affiliations

¹ Department of Biomedical Engineering, Key Laboratory of Biomedical Engineering of Ministry of Education, State Key Laboratory of Modern Optical Instrumentation, Zhejiang Provincial Key Laboratory of Cardio-Cerebral Vascular Detection Technology and Medicinal Effectiveness Appraisal, Zhejiang University, Hangzhou, China. yingkexu@zju.edu.cn.
² Department of Biomedical Engineering, Key Laboratory of Biomedical Engineering of Ministry of Education, State Key Laboratory of Modern Optical Instrumentation, Zhejiang Provincial Key Laboratory of Cardio-Cerebral Vascular Detection Technology and Medicinal Effectiveness Appraisal, Zhejiang University, Hangzhou, China.
³ Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA. derek.toomre@yale.edu.

PMID: 35819765
DOI: 10.1007/978-1-0716-2209-4_12

Abstract

Total internal reflection fluorescence microscopy (TIRFM) provides extremely thin optical sectioning with excellent signal-to-noise ratios, which allows for visualization of membrane dynamics at the cell surface with superb spatiotemporal resolution. In this chapter, TIRFM is used to record and analyze exocytosis of single glucose transporter-4 (GLUT4) containing vesicles in 3T3-L1 adipocytes.

Keywords: Exocytosis; GLUT4; Live-cell imaging; TIRFM; Vesicle; Vesicle fusion.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

3T3-L1 Cells
Adipocytes*
Animals
Cell Membrane / metabolism
Exocytosis*
Mice
Microscopy, Fluorescence / methods