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Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy
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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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“Compared to spontaneous Raman imaging with typical pixel dwell times of several seconds181920, pixel dwell times in CARS imaging are in the range of microseconds21 and thus significantly faster due to a 105 times higher CARS signal compared to spontaneous Raman scattering22.”
From Label-free in vivo analysis of intracellular lipid droplets in the oleaginous microalga Monoraphidium neglectum by coherent Raman scattering microscopy (2016) · cites this paper as [21]
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Metadata
- Authors
- Evans, C. L., Potma, E. O., Puoris'haag, M., Côté, D., Lin, C. P., & Xie, X. S.
- Affiliations
- Harvard University (US)
- Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138; and Wellman Center for Photomedicine, Massachusetts General Hospital, 55 Fruit Street, Boston, MA 02114
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2005-11
- Volume
- 102
- Issue
- 46
- Pages
- 16807-16812
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC1283840
- OpenAlex
- W2148691803
- Cited by
- 957
- References
- 27
- Citations by year
- 2026: 13 · 2025: 29 · 2024: 32 · 2023: 42 · 2022: 38 · 2021: 40 · 2020: 31 · 2019: 45 · 2018: 36 · 2017: 47 · 2016: 70 · 2015: 58 · 2014: 59 · 2013: 54 · 2012: 64
- FWCI
- 21.82
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Spectroscopy Techniques in Biomedical and Chemical Research
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Optical Imaging and Spectroscopy Techniques
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Spectroscopy and Quantum Chemical Studies
- Keywords
- coherent anti-Stokes Raman scattering microscopy, in vivo tissue imaging, vibrational imaging, noninvasive molecular imaging, skin, sebaceous glands, corneocytes, adipocytes, mouse skin
- MeSH terms
- Animals, Diffusion, Female, Mice, Inbred BALB C, Mice, Hairless, Microscopy, Mineral Oil, Scattering, Radiation, Skin, Spectrum Analysis, Raman, Mice
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