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      Coherent Raman scattering microscopy: capable solution in search of a larger audience

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          Abstract.

          Significance: Coherent Raman scattering (CRS) microscopy is an optical imaging technique with capabilities that could benefit a broad range of biomedical research studies.

          Aim: We reflect on the birth, rapid rise, and inescapable growing pains of the technique and look back on nearly four decades of developments to examine where the CRS imaging approach might be headed in the next decade to come.

          Approach: We provide a brief historical account of CRS microscopy, followed by a discussion of the challenges to disseminate the technique to a larger audience. We then highlight recent progress in expanding the capabilities of the CRS microscope and assess its current appeal as a practical imaging tool.

          Results: New developments in Raman tagging have improved the specificity and sensitivity of the CRS technique. In addition, technical advances have led to CRS microscopes that can capture hyperspectral data cubes at practical acquisition times. These improvements have broadened the application space of the technique.

          Conclusion: The technical performance of the CRS microscope has improved dramatically since its inception, but these advances have not yet translated into a substantial user base beyond a strong core of enthusiasts. Nonetheless, new developments are poised to move the unique capabilities of the technique into the hands of more users.

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          Most cited references91

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          Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering

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            Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy.

            Imaging living organisms with molecular selectivity typically requires the introduction of specific labels. Many applications in biology and medicine, however, would significantly benefit from a noninvasive imaging technique that circumvents such exogenous probes. In vivo microscopy based on vibrational spectroscopic contrast offers a unique approach for visualizing tissue architecture with molecular specificity. We have developed a sensitive technique for vibrational imaging of tissues by combining coherent anti-Stokes Raman scattering (CARS) with video-rate microscopy. Backscattering of the intense forward-propagating CARS radiation in tissue gives rise to a strong epi-CARS signal that makes in vivo imaging possible. This substantially large signal allows for real-time monitoring of dynamic processes, such as the diffusion of chemical compounds, in tissues. By tuning into the CH(2) stretching vibrational band, we demonstrate CARS imaging and spectroscopy of lipid-rich tissue structures in the skin of a live mouse, including sebaceous glands, corneocytes, and adipocytes, with unprecedented contrast at subcellular resolution.
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              Video-rate molecular imaging in vivo with stimulated Raman scattering.

              Optical imaging in vivo with molecular specificity is important in biomedicine because of its high spatial resolution and sensitivity compared with magnetic resonance imaging. Stimulated Raman scattering (SRS) microscopy allows highly sensitive optical imaging based on vibrational spectroscopy without adding toxic or perturbative labels. However, SRS imaging in living animals and humans has not been feasible because light cannot be collected through thick tissues, and motion-blur arises from slow imaging based on backscattered light. In this work, we enable in vivo SRS imaging by substantially enhancing the collection of the backscattered signal and increasing the imaging speed by three orders of magnitude to video rate. This approach allows label-free in vivo imaging of water, lipid, and protein in skin and mapping of penetration pathways of topically applied drugs in mice and humans.
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                Author and article information

                Contributors
                Journal
                J Biomed Opt
                J Biomed Opt
                JBOPFO
                JBO
                Journal of Biomedical Optics
                Society of Photo-Optical Instrumentation Engineers
                1083-3668
                1560-2281
                3 June 2021
                June 2021
                3 June 2021
                : 26
                : 6
                : 060601
                Affiliations
                [a ]University of California, Irvine , Department of Biomedical Engineering, Irvine, California, United States
                [b ]University of California, Irvine , Department of Chemistry, Irvine, California, United States
                Author notes
                [* ]Address all correspondence to Eric O. Potma, epotma@ 123456uci.edu
                Author information
                https://orcid.org/0000-0003-1199-9465
                https://orcid.org/0000-0003-3916-6131
                Article
                JBO-210102-PER 210102-PER
                10.1117/1.JBO.26.6.060601
                8174578
                34085436
                f6275216-01c2-413b-bd04-88cd110da1f9
                © 2021 The Authors

                Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

                History
                : 6 April 2021
                : 20 May 2021
                Page count
                Figures: 2, Tables: 0, References: 91, Pages: 11
                Funding
                Funded by: National Institutes of Health
                Award ID: R01-GM132506
                Categories
                Perspectives
                Paper
                Custom metadata
                Prince and Potma: Coherent Raman scattering microscopy…

                Biomedical engineering
                coherent raman scattering microscopy,optical imaging,lipid metabolism

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