17 December 2015 D38-cholesterol as a Raman active probe for imaging intracellular cholesterol storage
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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.
© 2015 Society of Photo-Optical Instrumentation Engineers (SPIE)
Alba Alfonso García, Simon G. Pfisterer, Howard Riezman, Elina Ikonen, Eric O. Potma, "D38-cholesterol as a Raman active probe for imaging intracellular cholesterol storage," Journal of Biomedical Optics 21(6), 061003 (17 December 2015). https://doi.org/10.1117/1.JBO.21.6.061003 . Submission:
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