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Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy.
- Source :
-
Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2019 Apr 02; Vol. 116 (14), pp. 6608-6617. Date of Electronic Publication: 2019 Mar 14. - 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.<br />Competing Interests: The authors declare no conflict of interest.
- Subjects :
- Animals
Cell Line, Tumor
Female
Humans
Mice
Spectrum Analysis, Raman
Spheroids, Cellular metabolism
Spheroids, Cellular pathology
Brain Neoplasms diagnostic imaging
Brain Neoplasms metabolism
Cone-Beam Computed Tomography
Glioblastoma diagnosis
Glioblastoma metabolism
Neoplasms, Experimental diagnostic imaging
Neoplasms, Experimental metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1091-6490
- Volume :
- 116
- Issue :
- 14
- Database :
- MEDLINE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Publication Type :
- Academic Journal
- Accession number :
- 30872474
- Full Text :
- https://doi.org/10.1073/pnas.1813044116