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Fluorescence Cross-Correlation Spectroscopy (FCCS) in Living Cells

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Fluorescence Spectroscopy and Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1076))

Abstract

Fluorescence cross-correlation spectroscopy (FCCS) is a single-molecule sensitive technique to quantitatively study interactions among fluorescently tagged biomolecules. Besides the initial implementation as dual-color FCCS (DC-FCCS), FCCS has several powerful derivatives, including single-wavelength FCCS (SW-FCCS), two-photon FCCS (TP-FCCS), and pulsed interleaved excitation FCCS (PIE-FCCS). However, to apply FCCS successfully, one needs to be familiar with procedures ranging from fluorescent labeling, instrumentation setup and alignment, sample preparation, and data analysis. Here, we describe the procedures to apply FCCS in various biological samples ranging from live cells to in vivo measurements, with the focus on DC-FCCS and SW-FCCS.

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Author information

Authors and Affiliations

  1. Faculty of Science, Department of Chemistry, National University of Singapore, Singapore, Singapore

    Xiaoxiao Ma

  2. Department of Chemistry, Faculty of Science, National University of Singapore, Singapore, Singapore

    Yong Hwee Foo

  3. Mechanobiology Institute, Singapore, Singapore

    Yong Hwee Foo

  4. Faculty of Science, Department of Chemistry and Department of Biological Sciences, National University of Singapore, Singapore, Singapore

    Thorsten Wohland

Authors
  1. Xiaoxiao Ma
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  2. Yong Hwee Foo
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  3. Thorsten Wohland
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Editor information

Editors and Affiliations

  1. KULeuven-Biomolecular Dynamics BioSCENTer, Leuven, Belgium

    Yves Engelborghs

  2. MicroSpectroscopy Centre, University of Wageningen Biochemistry, Wageningen, The Netherlands

    Antonie J.W.G. Visser

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Ma, X., Foo, Y.H., Wohland, T. (2014). Fluorescence Cross-Correlation Spectroscopy (FCCS) in Living Cells. In: Engelborghs, Y., Visser, A. (eds) Fluorescence Spectroscopy and Microscopy. Methods in Molecular Biology, vol 1076. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-649-8_25

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  • DOI: https://doi.org/10.1007/978-1-62703-649-8_25

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-648-1

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