Special Section on Selected Topics in Biophotonics: Photoacoustic Tomography and Fiber-Based Lasers and Supercontinuum Sources

D38-cholesterol as a Raman active probe for imaging intracellular cholesterol storage

[+] Author Affiliations
Alba Alfonso-García

University of California, Irvine, Department of Biomedical Engineering, Irvine, California 92697, United States

Simon G. Pfisterer, Elina Ikonen

University of Helsinki, Finland and Minerva Foundation Institute for Medical Research, Department of Anatomy and Research Programs Unit, Faculty of Medicine, Helsinki 00290, Finland

Howard Riezman

University of Geneva, NCCR Chemical Biology, Department of Biochemistry, CH-1211 Geneva, Switzerland

Eric O. Potma

University of California, Irvine, Department of Chemistry, Irvine, California 92697, United States

J. Biomed. Opt. 21(6), 061003 (Dec 17, 2015). doi:10.1117/1.JBO.21.6.061003
History: Received August 17, 2015; Accepted October 15, 2015
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Abstract.  We generated a highly deuterated cholesterol analog (D38-cholesterol) and demonstrated its use for selective vibrational imaging of cholesterol storage in mammalian cells. D38-cholesterol produces detectable signals in stimulated Raman scattering (SRS) imaging, is rapidly taken up by cells, and is efficiently metabolized by acyl-CoA cholesterol acyltransferase to form cholesteryl esters. Using hyperspectral SRS imaging of D38-cholesterol, we visualized cholesterol storage in lipid droplets. We found that some lipid droplets accumulated preferentially unesterified D38-cholesterol, whereas others stored D38-cholesteryl esters. In steroidogenic cells, D38-cholesteryl esters and triacylglycerols were partitioned into distinct sets of lipid droplets. Thus, hyperspectral SRS imaging of D38-cholesterol demonstrates a heterogeneous incorporation of neutral lipid species, i.e., free cholesterol, cholesteryl esters, and triacylglycerols, between individual lipid droplets in a cell.

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© 2015 Society of Photo-Optical Instrumentation Engineers

Citation

Alba Alfonso-García ; Simon G. Pfisterer ; Howard Riezman ; Elina Ikonen and Eric O. Potma
"D38-cholesterol as a Raman active probe for imaging intracellular cholesterol storage", J. Biomed. Opt. 21(6), 061003 (Dec 17, 2015). ; http://dx.doi.org/10.1117/1.JBO.21.6.061003


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