Scanning FCS for the Characterization of Protein Dynamics in Live Cells

Publikation: Beitrag in Buch/Konferenzbericht/Sammelband/GutachtenBeitrag in Buch/Sammelband/GutachtenBeigetragenBegutachtung

Beitragende

  • Zdeněk Petrášek - , Technische Universität Dresden (Autor:in)
  • Jonas Ries - , Technische Universität Dresden (Autor:in)
  • Petra Schwille - , Professur für Biophysik (Autor:in)

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

OriginalspracheEnglisch
TitelMethods in Enzymology
Herausgeber (Verlag)Elsevier Academic Press Inc
Seiten317-343
Seitenumfang27
PublikationsstatusVeröffentlicht - Jan. 2010
Peer-Review-StatusJa

Publikationsreihe

ReiheMethods in Enzymology
Band472
ISSN0076-6879

Externe IDs

PubMed 20580970

Schlagworte

ASJC Scopus Sachgebiete