Unit

UNIT 4.24 Fluorescence Correlation Spectroscopy in Living Cells: A Practical Approach

  1. Nihal Altan-Bonnet1,
  2. Grégoire Altan-Bonnet2

Published Online: 1 DEC 2009

DOI: 10.1002/0471143030.cb0424s45

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Altan-Bonnet, N. and Altan-Bonnet, G. 2009. Fluorescence Correlation Spectroscopy in Living Cells: A Practical Approach. Current Protocols in Cell Biology. 45:4.24:4.24.1–4.24.14.

Author Information

  1. 1

    Department of Biology, Rutgers University, Newark, New Jersey

  2. 2

    Program in Computational Biology and Immunology, Memorial Sloan Kettering Cancer Center, New York, New York

Publication History

  1. Published Online: 1 DEC 2009
  2. Published Print: DEC 2009

Abstract

Fluorescence correlation spectroscopy (FCS) is a single-molecule fluorescence technique used to monitor molecular dynamics. It can be applied to living cells expressing fluorescently labeled proteins and lipids to determine the diffusion timescales and the total number of diffusing fluorescent molecules in the cell. A practical step-by-step approach to performing FCS with a commercial spectroscopy/microscopy system, the Zeiss Confocor 3, how to set up live-cell FCS experiments, acquire reliable data, and finally how to analyze the data acquired, are all described in this unit. Curr. Protoc. Cell Biol. 45:4.24.1-4.24.14. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • fluorescence correlation spectroscopy;
  • FCS;
  • live-cell imaging;
  • green fluorescent protein;
  • GFP;
  • diffusion