Analysis of Cholesterol Trafficking with Fluorescent Probes

Frederick R. Maxfield, Daniel Wustner

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

Abstract

Cholesterol plays an important role in determining the biophysical properties of biological membranes, and its concentration is tightly controlled by homeostatic processes. The intracellular transport of cholesterol among organelles is a key part of the homeostatic mechanism, but sterol transport processes are not well understood. Fluorescence microscopy is a valuable tool for studying intracellular transport processes, but this method can be challenging for lipid molecules because addition of a fluorophore may alter the properties of the molecule greatly. We discuss the use of fluorescent molecules that can bind to cholesterol to reveal its distribution in cells. We also discuss the use of intrinsically fluorescent sterols that closely mimic cholesterol, as well as some minimally modified fluorophore-labeled sterols. Methods for imaging these sterols by conventional fluorescence microscopy and by multiphoton microscopy are described. Some label-free methods for imaging cholesterol itself are also discussed briefly.
Original languageEnglish
Title of host publicationLIPIDS, VOL 108
EditorsG DiPaolo, MR Wenk
Number of pages27
Volume108
Publication date2012
Pages367-393
DOIs
Publication statusPublished - 2012
SeriesMethods in Cell Biology
Volume108
ISSN0091-679X

Cite this

Maxfield, F. R., & Wustner, D. (2012). Analysis of Cholesterol Trafficking with Fluorescent Probes. In G. DiPaolo, & MR. Wenk (Eds.), LIPIDS, VOL 108 (Vol. 108, pp. 367-393). Methods in Cell Biology, Vol.. 108 https://doi.org/10.1016/B978-0-12-386487-1.00017-1
Maxfield, Frederick R. ; Wustner, Daniel. / Analysis of Cholesterol Trafficking with Fluorescent Probes. LIPIDS, VOL 108. editor / G DiPaolo ; MR Wenk. Vol. 108 2012. pp. 367-393 (Methods in Cell Biology, Vol. 108).
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Maxfield, FR & Wustner, D 2012, Analysis of Cholesterol Trafficking with Fluorescent Probes. in G DiPaolo & MR Wenk (eds), LIPIDS, VOL 108. vol. 108, Methods in Cell Biology, vol. 108, pp. 367-393. https://doi.org/10.1016/B978-0-12-386487-1.00017-1

Analysis of Cholesterol Trafficking with Fluorescent Probes. / Maxfield, Frederick R.; Wustner, Daniel.

LIPIDS, VOL 108. ed. / G DiPaolo; MR Wenk. Vol. 108 2012. p. 367-393 (Methods in Cell Biology, Vol. 108).

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

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AB - Cholesterol plays an important role in determining the biophysical properties of biological membranes, and its concentration is tightly controlled by homeostatic processes. The intracellular transport of cholesterol among organelles is a key part of the homeostatic mechanism, but sterol transport processes are not well understood. Fluorescence microscopy is a valuable tool for studying intracellular transport processes, but this method can be challenging for lipid molecules because addition of a fluorophore may alter the properties of the molecule greatly. We discuss the use of fluorescent molecules that can bind to cholesterol to reveal its distribution in cells. We also discuss the use of intrinsically fluorescent sterols that closely mimic cholesterol, as well as some minimally modified fluorophore-labeled sterols. Methods for imaging these sterols by conventional fluorescence microscopy and by multiphoton microscopy are described. Some label-free methods for imaging cholesterol itself are also discussed briefly.

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Maxfield FR, Wustner D. Analysis of Cholesterol Trafficking with Fluorescent Probes. In DiPaolo G, Wenk MR, editors, LIPIDS, VOL 108. Vol. 108. 2012. p. 367-393. (Methods in Cell Biology, Vol. 108). https://doi.org/10.1016/B978-0-12-386487-1.00017-1