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Journal of Biomedical Optics

Integrated femtosecond stimulated Raman scattering and two-photon fluorescence imaging of subcellular lipid and vesicular structures
Author(s): Xuesong Li; Wen Jiun Lam; Zhe Cao; Yan Hao; Qiqi Sun; Sicong He; Ho Yi Mak; Jianan Y. Qu
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Paper Abstract

The primary goal of this study is to demonstrate that stimulated Raman scattering (SRS) as a new imaging modality can be integrated into a femtosecond (fs) nonlinear optical (NLO) microscope system. The fs sources of high pulse peak power are routinely used in multimodal nonlinear microscopy to enable efficient excitation of multiple NLO signals. However, with fs excitations, the SRS imaging of subcellular lipid and vesicular structures encounters significant interference from proteins due to poor spectral resolution and a lack of chemical specificity, respectively. We developed a unique NLO microscope of fs excitation that enables rapid acquisition of SRS and multiple two-photon excited fluorescence (TPEF) signals. In the <italic<in vivo</italic< imaging of transgenic <italic<C. elegans</italic< animals, we discovered that by cross-filtering false positive lipid signals based on the TPEF signals from tryptophan-bearing endogenous proteins and lysosome-related organelles, the imaging system produced highly accurate assignment of SRS signals to lipid. Furthermore, we demonstrated that the multimodal NLO microscope system could sequentially image lipid structure/content and organelles, such as mitochondria, lysosomes, and the endoplasmic reticulum, which are intricately linked to lipid metabolism.

Paper Details

Date Published: 18 November 2015
PDF: 4 pages
J. Biomed. Opt. 20(11) 110501 doi: 10.1117/1.JBO.20.11.110501
Published in: Journal of Biomedical Optics Volume 20, Issue 11
Show Author Affiliations
Xuesong Li, Hong Kong Univ. of Science and Technology (China)
Wen Jiun Lam, Hong Kong Univ. of Science and Technology (China)
Zhe Cao, Hong Kong Univ. of Science and Technology (China)
Yan Hao, Hong Kong Univ. of Science and Technology (China)
Qiqi Sun, Hong Kong Univ. of Science and Technology (China)
Sicong He, Hong Kong Univ. of Science and Technology (China)
Ho Yi Mak, Hong Kong Univ. of Science and Technology (China)
Jianan Y. Qu, Hong Kong Univ. of Science and Technology (China)


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