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Single Molecule Analysis pp 93–113Cite as

A Brief Introduction to Single-Molecule Fluorescence Methods

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1665))

Abstract

One of the more popular single-molecule approaches in biological science is single-molecule fluorescence microscopy, which will be the subject of the following section of this volume. Fluorescence methods provide the sensitivity required to study biology on the single-molecule level, but they also allow access to useful measurable parameters on time and length scales relevant for the biomolecular world. Before several detailed experimental approaches will be addressed, we will first give a general overview of single-molecule fluorescence microscopy. We start with discussing the phenomenon of fluorescence in general and the history of single-molecule fluorescence microscopy. Next, we will review fluorescent probes in more detail and the equipment required to visualize them on the single-molecule level. We will end with a description of parameters measurable with such approaches, ranging from protein counting and tracking, single-molecule localization super-resolution microscopy, to distance measurements with Förster Resonance Energy Transfer and orientation measurements with fluorescence polarization.

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Authors and Affiliations

  1. LaserLaB and Department of Physics and Astronomy, Vrije Universiteit, De Boelelaan 1081, 1081 HV, Amsterdam, The Netherlands

    Siet M. J. L. van den Wildenberg, Bram Prevo & Erwin J. G. Peterman

  2. Équipe de Volcanologie, Observatoire de Physique de Globe, Clermant-Ferrand, France

    Siet M. J. L. van den Wildenberg

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  1. Siet M. J. L. van den Wildenberg
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  2. Bram Prevo
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  3. Erwin J. G. Peterman
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Correspondence to Erwin J. G. Peterman .

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  1. Department of Physics & Astronomy, Vrije Universiteit, Amsterdam, The Netherlands

    Erwin J. G. Peterman

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van den Wildenberg, S.M.J.L., Prevo, B., Peterman, E.J.G. (2018). A Brief Introduction to Single-Molecule Fluorescence Methods. In: Peterman, E. (eds) Single Molecule Analysis. Methods in Molecular Biology, vol 1665. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7271-5_5

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  • DOI: https://doi.org/10.1007/978-1-4939-7271-5_5

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