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Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy.

Authors :
Mian Wei
Lingyan Shi
Yihui Shen
Zhilun Zhao
Guzman, Asja
Kaufman, Laura J.
Lu Wei
Wei Min
Source :
Proceedings of the National Academy of Sciences of the United States of America. 4/2/2019, Vol. 116 Issue 14, p6608-6617. 10p.
Publication Year :
2019

Abstract

Three-dimensional visualization of tissue structures using optical microscopy facilitates the understanding of biological functions. However, optical microscopy is limited in tissue penetration due to severe light scattering. Recently, a series of tissue-clearing techniques have emerged to allow significant depth-extension for fluorescence imaging. Inspired by these advances, we develop a volumetric chemical imaging technique that couples Raman-tailored tissue-clearing with stimulated Raman scattering (SRS) microscopy. Compared with the standard SRS, the clearing-enhanced SRS achieves greater than 10-times depth increase. Based on the extracted spatial distribution of proteins and lipids, our method reveals intricate 3D organizations of tumor spheroids, mouse brain tissues, and tumor xenografts. We further develop volumetric phasor analysis of multispectral SRS images for chemically specific clustering and segmentation in 3D. Moreover, going beyond the conventional label-free paradigm, we demonstrate metabolic volumetric chemical imaging, which allows us to simultaneously map out metabolic activities of protein and lipid synthesis in glioblastoma. Together, these results support volumetric chemical imaging as a valuable tool for elucidating comprehensive 3D structures, compositions, and functions in diverse biological contexts, complementing the prevailing volumetric fluorescence microscopy. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
00278424
Volume :
116
Issue :
14
Database :
Academic Search Index
Journal :
Proceedings of the National Academy of Sciences of the United States of America
Publication Type :
Academic Journal
Accession number :
135759809
Full Text :
https://doi.org/10.1073/pnas.1813044116