Assessing cholesterol storage in live cells and C. elegans by stimulated Raman scattering imaging of phenyl-Diyne cholesterol

Hyeon Jeong Lee; Wandi Zhang; Delong Zhang; Yang Yang; Bin Liu; Eric L Barker; Kimberly K Buhman; Lyudmila V Slipchenko; Mingji Dai; Ji-Xin Cheng

doi:10.1038/srep07930

Assessing cholesterol storage in live cells and C. elegans by stimulated Raman scattering imaging of phenyl-Diyne cholesterol

Sci Rep. 2015 Jan 22:5:7930. doi: 10.1038/srep07930.

Authors

Affiliations

¹ 1] Interdisciplinary Life Science Program, Purdue University, West Lafayette, IN, USA [2] Department of Comparative Pathobiology, Purdue University, West Lafayette, IN, USA.
² Department of Chemistry, Purdue University, West Lafayette, IN, USA.
³ National Key Laboratory of Science and Technology on Tunable Laser, Harbin Institute of Technology, Harbin 150080, China.
⁴ 1] Interdisciplinary Life Science Program, Purdue University, West Lafayette, IN, USA [2] Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN, USA.
⁵ 1] Interdisciplinary Life Science Program, Purdue University, West Lafayette, IN, USA [2] Department of Nutrition Science, Purdue University, West Lafayette, IN, USA.
⁶ 1] Interdisciplinary Life Science Program, Purdue University, West Lafayette, IN, USA [2] Department of Chemistry, Purdue University, West Lafayette, IN, USA.
⁷ 1] Interdisciplinary Life Science Program, Purdue University, West Lafayette, IN, USA [2] Department of Chemistry, Purdue University, West Lafayette, IN, USA [3] Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA.

Abstract

We report a cholesterol imaging method using rationally synthesized phenyl-diyne cholesterol (PhDY-Chol) and stimulated Raman scattering (SRS) microscope. The phenyl-diyne group is biologically inert and provides a Raman scattering cross section that is 88 times larger than the endogenous C = O stretching mode. SRS microscopy offers an imaging speed that is faster than spontaneous Raman microscopy by three orders of magnitude, and a detection sensitivity of 31 μM PhDY-Chol (~1,800 molecules in the excitation volume). Inside living CHO cells, PhDY-Chol mimics the behavior of cholesterol, including membrane incorporation and esterification. In a cellular model of Niemann-Pick type C disease, PhDY-Chol reflects the lysosomal accumulation of cholesterol, and shows relocation to lipid droplets after HPβCD treatment. In live C. elegans, PhDY-Chol mimics cholesterol uptake by intestinal cells and reflects cholesterol storage. Together, our work demonstrates an enabling platform for study of cholesterol storage and trafficking in living cells and vital organisms.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
CHO Cells
Caenorhabditis elegans / chemistry
Caenorhabditis elegans / metabolism
Cholesterol / chemical synthesis
Cholesterol / isolation & purification
Cholesterol / metabolism*
Cricetulus
Diynes / chemical synthesis
Diynes / chemistry
Lipid Droplets / chemistry
Lipid Droplets / metabolism
Lysosomes / metabolism*
Lysosomes / pathology
Molecular Imaging*
Niemann-Pick Disease, Type C / metabolism*
Niemann-Pick Disease, Type C / pathology
Spectrum Analysis, Raman

Substances

Diynes
Cholesterol

Abstract

Publication types

MeSH terms

Substances

Grants and funding