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

Proc Natl Acad Sci U S A. 2005 Nov 15;102(46):16807-12. doi: 10.1073/pnas.0508282102. Epub 2005 Nov 1.

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 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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Diffusion
  • Female
  • Mice
  • Mice, Hairless
  • Mice, Inbred BALB C
  • Microscopy / methods*
  • Mineral Oil / metabolism
  • Scattering, Radiation
  • Skin / metabolism
  • Spectrum Analysis, Raman / methods*

Substances

  • Mineral Oil