title: Assessing cholesterol storage in live cells and C. elegans by stimulated Raman scattering imaging of phenyl-Diyne cholesterol.
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.
authors.affiliation: 1] Interdisciplinary Life Science Program, Purdue University, West Lafayette, IN, USA [2] Department of Comparative Pathobiology, Purdue University, West Lafayette, IN, USA.
authors.forename: Hyeon Jeong
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, IN, USA.
authors.forename: Wandi
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, IN, USA.
authors.forename: Delong
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, IN, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.affiliation: National Key Laboratory of Science and Technology on Tunable Laser, Harbin Institute of Technology, Harbin 150080, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
authors.affiliation: 1] Interdisciplinary Life Science Program, Purdue University, West Lafayette, IN, USA [2] Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN, USA.
authors.forename: Eric L
authors.initials: EL
authors.lastname: Barker
authors.affiliation: 1] Interdisciplinary Life Science Program, Purdue University, West Lafayette, IN, USA [2] Department of Nutrition Science, Purdue University, West Lafayette, IN, USA.
authors.forename: Kimberly K
authors.initials: KK
authors.lastname: Buhman
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, IN, USA.
authors.forename: Lyudmila V
authors.initials: LV
authors.lastname: Slipchenko
authors.affiliation: 1] Interdisciplinary Life Science Program, Purdue University, West Lafayette, IN, USA [2] Department of Chemistry, Purdue University, West Lafayette, IN, USA.
authors.forename: Mingji
authors.initials: M
authors.lastname: Dai
authors.affiliation: 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.
authors.forename: Ji-Xin
authors.initials: JX
authors.lastname: Cheng
mesh_terms: D000818:Animals; D016466:CHO Cells; D017173:Caenorhabditis elegans / Q000737:chemistry / Q000378:metabolism; D002784:Cholesterol / Q000138:chemical synthesis / Q000302:isolation & purification / Q000378:metabolism*; D003412:Cricetulus; D053280:Diynes / Q000138:chemical synthesis / Q000737:chemistry; D066292:Lipid Droplets / Q000737:chemistry / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism* / Q000473:pathology; D057054:Molecular Imaging; D052556:Niemann-Pick Disease, Type C / Q000378:metabolism* / Q000473:pathology; D013059:Spectrum Analysis, Raman
chemical_list: D053280:Diynes; D002784:Cholesterol