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

November 1, 2005
102 (46) 16807-16812

Abstract

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 CH2 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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Acknowledgments

We thank Dr. David Bell for help with tissue sectioning. This research was funded by the National Institutes of Health, the National Science Foundation, the Department of Defense Medical Free Electron Program, and in particular the National Institutes of Health Director's Pioneer Award (to X.S.X.). C.L.E. was supported by a National Science Foundation Graduate Research Fellowship.

Supporting Information

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Information & Authors

Information

Published in

Go to Proceedings of the National Academy of Sciences
Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 102 | No. 46
November 15, 2005
PubMed: 16263923

Classifications

Submission history

Received: August 18, 2005
Published online: November 1, 2005
Published in issue: November 15, 2005

Keywords

  1. nonlinear microscopy
  2. vibrational imaging
  3. back scattering

Acknowledgments

We thank Dr. David Bell for help with tissue sectioning. This research was funded by the National Institutes of Health, the National Science Foundation, the Department of Defense Medical Free Electron Program, and in particular the National Institutes of Health Director's Pioneer Award (to X.S.X.). C.L.E. was supported by a National Science Foundation Graduate Research Fellowship.

Authors

Affiliations

Conor L. Evans
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
Eric O. Potma
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
Mehron Puoris'haag
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
Daniel Côté
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
Charles P. Lin§
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
X. Sunney Xie§
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

Notes

§
To whom correspondence may be addressed. E-mail: [email protected] or [email protected].
Present address: Department of Chemistry, University of California, Irvine, CA 92697.
Communicated by William Klemperer, Harvard University, Cambridge, MA, September 22, 2005

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    Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy
    Proceedings of the National Academy of Sciences
    • Vol. 102
    • No. 46
    • pp. 16531-16905

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