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The Nucleus pp 321-341 | Cite as

Fluorescence Correlation Spectroscopy to Assess the Mobility of Nuclear Proteins

  • Stefanie Weidtkamp-Peters
  • Klaus Weisshart
  • Lars Schmiedeberg
  • Peter Hemmerich
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 464)

Abstract

Recent developments in cell biology and microscopy techniques enable us to observe macromolecular assemblies in their natural setting: the living cell. These emerging technologies have revealed novel concepts in nuclear cell biology. In order to further elucidate the biochemistry of gene expression, replication, and genome maintenance, the major challenge is now to precisely determine the dynamics of nuclear proteins in the context of the structural organization of the nucleus. Fluorescence correlation spectroscopy (FCS) is an attractive alternative to photobleaching and photoactivation techniques for the analysis of protein dynamics at single-molecule resolution. Here we describe how FCS can be applied to retrieve biophysical parameters of nuclear proteins in living cells.

Keywords

Cell nucleus Obstructed diffusion Kinetic microscopy Living cell Diffusion coefficient Enhanced green fluorescent protein Bleaching 

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Copyright information

© Humana Press, a part of Springer Science + Business Media, LLC 2008

Authors and Affiliations

  • Stefanie Weidtkamp-Peters
    • 1
  • Klaus Weisshart
    • 2
  • Lars Schmiedeberg
    • 3
  • Peter Hemmerich
    • 1
  1. 1.Leibniz Institute for Age ResearchFritz Lipmann InstituteJenaGermany
  2. 2.Carl Zeiss JenaJenaGermany
  3. 3.Wellcome Trust Center for Cell Biology, Institute of Cell and Molecular BiologyUniversity of EdinburghEdinburghScotland, UK

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