Scanning FCS for the Characterization of Protein Dynamics in Live Cells

Research output: Contribution to book/Conference proceedings/Anthology/ReportChapter in book/Anthology/ReportContributedpeer-review

Contributors

  • Zdeněk Petrášek - , TUD Dresden University of Technology (Author)
  • Jonas Ries - , TUD Dresden University of Technology (Author)
  • Petra Schwille - , Chair of Biophysics (Author)

Abstract

Scanning fluorescence correlation spectroscopy (sFCS) is the generic term for a group of fluorescence correlation techniques where the measurement volume is moved across the sample in a defined way. The introduction of scanning is motivated by its ability to alleviate or remove several distinct problems often encountered in standard FCS, and thus, to extend the range of applicability of fluorescence correlation methods in biological systems. These problems include poor statistical accuracy in measurements with slowly moving molecules, photobleaching, optical distortions affecting the calibration of the measurement volume, membrane instabilities, etc. Here, we present an overview of sFCS methods, explaining their benefits, implementation details, requirements, and limitations, as well as relations to each other. Further, we give examples of different sFCS implementations as applied to cellular systems, namely large-circle sFCS to measure protein dynamics in embryo cortex and line sFCS to measure protein diffusion and interactions in unstable membranes.

Details

Original languageEnglish
Title of host publicationMethods in Enzymology
PublisherElsevier Academic Press Inc
Pages317-343
Number of pages27
Publication statusPublished - Jan 2010
Peer-reviewedYes

Publication series

SeriesMethods in Enzymology
Volume472
ISSN0076-6879

External IDs

PubMed 20580970

Keywords

ASJC Scopus subject areas