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Membrane Biophysics pp 147–173Cite as

Analysis and Applications of Single-Molecule Fluorescence in Live Cell Membranes

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Abstract

In the past decades, single-molecule fluorescence (SMF) techniques have been booming as they provide researchers with valuable molecular information that are unobtainable by using ensemble techniques. Applications of SMF techniques to live cell membranes have revolutionized our views on many cellular processes. In this chapter, we describe the basics of SMF techniques such as SMF imaging, single-molecule fluorescence resonance energy transfer (sm-FRET), and fluorescence correlation spectroscopy (FCS) and then discuss their contributions to the understanding of live cell membranes. We finally provide the practical protocols of applying SMF techniques to live cell membranes, including the choice of fluorescent probes, labeling strategies, cell sample preparation, instrumentation setup, and data analysis.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 21573289) and the Natural Science Foundation of Shandong Province (No. ZR2014BM028).

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  1. State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, China

    Hua He, Xiaojuan Wang & Fang Huang

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  1. Hua He
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Correspondence to Fang Huang .

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  1. Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, China

    Hongda Wang

  2. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, Liaoning, China

    Guohui Li

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He, H., Wang, X., Huang, F. (2018). Analysis and Applications of Single-Molecule Fluorescence in Live Cell Membranes. In: Wang, H., Li, G. (eds) Membrane Biophysics. Springer, Singapore. https://doi.org/10.1007/978-981-10-6823-2_6

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