Journal
JOURNAL OF BIOMEDICAL OPTICS
Volume 21, Issue 6, Pages -Publisher
SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
DOI: 10.1117/1.JBO.21.6.061003
Keywords
biophotonics; microscopy; nonlinear optics; Raman spectroscopy
Funding
- NIH [P41-RR01192]
- Academy of Finland [272130, 282192, 284667, 275964]
- Sigrid Juselius Foundation
- Paulo Foundation
- Academy of Finland (AKA) [284667, 275964, 284667, 275964] Funding Source: Academy of Finland (AKA)
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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. (C) 2015 Society of Photo-Optical Instrumentation Engineers (SPIE)
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