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Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy

Wei, Mian and Shi, Lingyan and Shen, Yihui and Zhao, Zhilun and Guzman, Asja and Kaufman, Laura J. and Wei, Lu and Min, Wei (2019) Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy. Proceedings of the National Academy of Sciences of the United States of America, 116 (14). pp. 6608-6617. ISSN 0027-8424. https://resolver.caltech.edu/CaltechAUTHORS:20190314-130512034

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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1073/pnas.1813044116DOIArticle
https://www.pnas.org/content/suppl/2019/03/13/1813044116.DCSupplementalPublisherSupporting Information
ORCID:
AuthorORCID
Wei, Lu0000-0001-9170-2283
Min, Wei0000-0003-2570-3557
Additional Information:© 2019 National Academy of Sciences. Published under the PNAS license. Edited by Lance L. Munn, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA, and accepted by Editorial Board Member Rakesh K. Jain February 19, 2019 (received for review July 29, 2018). PNAS published ahead of print March 14, 2019. We thank E. Silveira for technical assistance; Y. Yang and X. Qu for discussion; and T. Swayne and the Confocal and Specialized Microscopy Shared Resource of the Herbert Irving Comprehensive Cancer Center at Columbia University for help with 3D image analysis, supported by NIH Grant P30 CA013696. W.M. acknowledges support of R01EB020892 from the NIH and the Camille and Henry Dreyfus Foundation. M.W. and L.S. contributed equally to this work. Author contributions: M.W., L.S., L.W., and W.M. designed research; M.W. and L.S. performed research; Y.S., Z.Z., A.G., and L.J.K. contributed new reagents/analytic tools; M.W. analyzed data; and M.W., L.S., L.W., and W.M. wrote the paper. The authors declare no conflict of interest. This article is a PNAS Direct Submission. L.L.M. is a guest editor invited by the Editorial Board. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1813044116/-/DCSupplemental.
Funders:
Funding AgencyGrant Number
NIHP30 CA013696
NIHR01EB020892
Camille and Henry Dreyfus FoundationUNSPECIFIED
Subject Keywords:volumetric imaging; stimulated Raman scattering; tissue clearing; metabolic imaging; cancer metabolism
Issue or Number:14
Record Number:CaltechAUTHORS:20190314-130512034
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20190314-130512034
Official Citation:Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy. Mian Wei, Lingyan Shi, Yihui Shen, Zhilun Zhao, Asja Guzman, Laura J. Kaufman, Lu Wei, Wei Min. Proceedings of the National Academy of Sciences Apr 2019, 116 (14) 6608-6617; DOI: 10.1073/pnas.1813044116
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:93811
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:14 Mar 2019 20:14
Last Modified:23 Oct 2019 20:57

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