In vivo study of lipid synthesis and lipolysis dynamics by stimulated Raman scattering microscopy

Xuesong Li; Yan Li; Sicong He; Congping Chen; Zhongya Qin; Ho Yi Mak; Jianan Y. Qu

doi:10.1117/12.2286773

23 February 2018 In vivo study of lipid synthesis and lipolysis dynamics by stimulated Raman scattering microscopy

Xuesong Li, Yan Li, Sicong He, Congping Chen, Zhongya Qin, Ho Yi Mak, Jianan Y. Qu

Proceedings Volume 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII; 104981Q (2018) https://doi.org/10.1117/12.2286773
Event: SPIE BiOS, 2018, San Francisco, California, United States

Abstract

To understand the mechanisms of important lipid-related biological processes and diseases, it is highly demanded to study the dynamics of lipids in living biological system with high spatiotemporal resolution. However, in vivo quantitative analysis of lipid synthesis and lipolysis has been technically difficult to achieve by conventional lipid extraction and fluorescent staining methods. Recently, SRS microscopy has emerged as a powerful tool to probe small molecules with alkyne (C≡C) or deuterium (C-D) labeling in cell-silent region. The Raman tags have been used for the quantitative study of lipids in cells. In this study, we investigated metabolic dynamics of lipid droplets (LDs) by tracing the alkyne-tagged fatty acid 17-ODYA and deuterium-labeled saturated and unsaturated fatty acids PA-D₃₁ & OA-D₃₄ in living C. elegans. Specifically, we developed a hyperspectral SRS microscope system for LDs characterization. The system can sequentially excite and probe the stimulated Raman scattering-induced CH₂ stretching of endogenous lipids information (2863 cm-1), C≡C stretching from 17-ODYA (2125 cm^-1) and C-D stretching from deuterium-labeled fatty acids (2117 cm^-1). We first examined the concentration levels of fatty acids in E. coli OP50. Two major lipid metabolic processes, namely uptake and turnover, were further studied in adult C. elegans. We imaged alkyne-tagged and deuterated fatty acids using SRS and traced their uptake, transportation, incorporation and turnover over time. Additionally, several other treatments including starvation were also conducted to study their effects on metabolic dynamics of pulse labeled 17-ODYA, PA-D₃₁ and OA-D₃₄.

Conference Presentation

Citation Download Citation

Xuesong Li, Yan Li, Sicong He, Congping Chen, Zhongya Qin, Ho Yi Mak, and Jianan Y. Qu "In vivo study of lipid synthesis and lipolysis dynamics by stimulated Raman scattering microscopy", Proc. SPIE 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII, 104981Q (23 February 2018); https://doi.org/10.1117/12.2286773

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