Scanning FCS for the Characterization of Protein Dynamics in Live Cells
Research output: Contribution to book/Conference proceedings/Anthology/Report › Chapter in book/Anthology/Report › Contributed › peer-review
Contributors
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 language | English |
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Title of host publication | Methods in Enzymology |
Publisher | Elsevier Academic Press Inc |
Pages | 317-343 |
Number of pages | 27 |
Publication status | Published - Jan 2010 |
Peer-reviewed | Yes |
Publication series
Series | Methods in Enzymology |
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Volume | 472 |
ISSN | 0076-6879 |
External IDs
PubMed | 20580970 |
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