Abstract
Fluorescence correlation spectroscopy (FCS) can add dynamic molecular information to images of live cells. For example, a confocal laser scanning microscope (CLSM) equipped with an accessory FCS unit provides the possibility to first image the spatial distribution of a fluorescent protein before probing its mobility within defined regions of interest. Whereas specific protein–protein interactions are preferably assayed with a dual-color approach, single-color FCS can still provide valuable information about the size of the diffusing entities and potential interactions with other, nonfluorescent, proteins or subcellular structures. Because number fluctuations are measured, the concentrations of freely diffusing complexes and their state of oligomerization are accessible.
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Acknowledgments
I thank Wolfgang Staroske for providing his Matlab fitting script, Karin Crell, and Ellen Sieber for their assistance in cell culture.
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Weidemann, T. (2014). Application of Fluorescence Correlation Spectroscopy (FCS) to Measure the Dynamics of Fluorescent Proteins in Living Cells. In: Engelborghs, Y., Visser, A. (eds) Fluorescence Spectroscopy and Microscopy. Methods in Molecular Biology, vol 1076. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-649-8_24
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DOI: https://doi.org/10.1007/978-1-62703-649-8_24
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