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.
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 (Presented at SPIE BiOS: January 30, 2018; Published: 23 February 2018); https://doi.org/10.1117/12.2286773.
Conference Presentations are recordings of oral presentations given at SPIE conferences and published as part of the proceedings. They include the speaker's narration with video of the slides and animations. Most include full-text papers. Interactive, searchable transcripts and closed captioning are now available for 2018 presentations, with transcripts for prior recordings added daily.
Search our growing collection of more than 16,000 conference presentations, including many plenaries and keynotes.