DO-SRS and MPF imaging of metabolic activities

Pegah Bagheri; Khang Hoang; Anthony Fung; Sahran Hussain; Lingyan Shi

doi:10.1117/12.2607975

2 March 2022 DO-SRS and MPF imaging of metabolic activities

Pegah Bagheri, Khang Hoang, Anthony Fung, Sahran Hussain, Lingyan Shi

Proceedings Volume 11973, Advanced Chemical Microscopy for Life Science and Translational Medicine 2022; 1197309 (2022) https://doi.org/10.1117/12.2607975
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

Emerging studies have shown that oxidative imbalance is critical in disease progression such as cancer and Alzheimer’s [1, 2]. This variation can lead to the upregulation of certain metabolic pathways inducing diseases and disorders. Aromatic amino acids (AAA) are involved with the production of Reactive Oxygen Species (ROS), resulting in the increase of oxidative stress [3]. AAA studies typically rely on gas chromatography (GC) or mass spectroscopy (MS)-based imaging techniques to study lipids; however, these methods lack the ability to show the cell’s lipid spatial distribution or require fluorescent dyes that can interfere with the cell’s molecular activities [4, 5]. Here, we established an optical imaging approach that combines D2O (heavy water) probed Stimulated Raman scattering (DO-SRS) and Multiphoton Fluorescence (MPF) microscopy to directly visualize metabolic activities in situ in cancer cells under the regulation of excess AAA, specifically Phenylalanine and Tryptophan. The cellular spatial distribution of de novo lipogenesis, unsaturated and saturated lipids, NADH, Flavin, and new protein synthesis were quantitatively imaged and examined. We discovered an increase in de novo lipogenesis, Flavin/(Flavin + NADH), and unsaturated to saturated lipids in the cancer cells treated with excess AAAs. Decrease of protein turnover rate occurred in the same treated cells with observations of higher lipid droplet content. These observed metabolic activities are signs of mitochondrial dysfunction and oxidative stress. Our study demonstrates that DO-SRS can be used as a high-resolution imaging platform to study AAA regulated metabolic activities in cells and elucidates the linkage between lipid metabolism and cancer.

Conference Presentation

Citation Download Citation

Pegah Bagheri, Khang Hoang, Anthony Fung, Sahran Hussain, and Lingyan Shi "DO-SRS and MPF imaging of metabolic activities", Proc. SPIE 11973, Advanced Chemical Microscopy for Life Science and Translational Medicine 2022, 1197309 (2 March 2022); https://doi.org/10.1117/12.2607975

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