Coherent Anti-Stokes Raman Scattering (CARS) microscopy

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Coherent Anti-Stokes Raman Scattering (CARS) microscopy

Principal investigator : Hervé Rigneault

An ultrasensitive technique without fluorophores

Fluorescence based microscopies are powerful tools to study the living world at the molecular scale. But their applications is often limited by dye phototoxicity and photobleaching. Furthermore any labeling procedure requires a complex chemistry and the fluorescent labeling of biological molecules is sometimes very difficult or impossible.

An alternative technique for sensitive detection is the vibrational spectroscopy that is not based on electronic excitations. Unfortunately standard vibrational spectroscopic methods do not offer high sensitivity (IR microscopy) or require high excitation intensities (Raman microscopy) that are not compatible with living systems.

Instrumental developments

Coherent anti-stokes Raman scattering microscopy is a modern pulse laser based technique which bypasses the problem of fluorescence and presents an intrinsic 3D resolution similar to two-photon confocal microscopy. In this process, the sample is shined both by a laser beam at pulsation wP and a stokes beam at pulsation wS. If wP-wS=wR, where wR is an intrinsic Raman vibration of a molecular species present in the sample, a four wave mixing process takes place which leads to the coherent generation of the anti-stokes pulsation at frequency 2wp-wS=wAS. Scanning the sample generates an image whose contrast comes from the CARS signal which is directly related to the local concentration of molecular bonds at the Raman frequency of interest.

We have implemented the first experimental setup in France and are currently working on the CARS detection of cellular pathologies.


Our team is strongly involved in the European research network CARSEXplorer, funded by the European Commission within its FP7 Specific programme Cooperation.

CARSExplorer official website

People involved

- Hervé Rigneault
- Patrick Ferrand
- Sophie Brustlein
- David Gachet
- Fabiana Munhoz

Selected publications

  • S. Brustlein, P. Ferrand, N. Walther, S. Brasselet, C. Billaudeau, D.
    Marguet and H Rigneault, Optical Parametric Oscillators based light source for Coherent Raman Scattering microscopy : a practical overview, J. Biomed. Opt. 16, 021106 (2011) - PDF
  • F. Munhoz, H. Rigneault, and S. Brasselet, High Order Symmetry Structural Properties of Vibrational Resonances Using Multiple-Field Polarization Coherent Anti-Stokes Raman Spectroscopy Microscopy, Phys. Rev. Lett., 105, 123903 (2010) - PDF
  • D. Gachet, S. Brustlein, and H. Rigneault, Revisiting the Young’s double slit experiment for background-free nonlinear Raman spectroscopy and microscopy, Phys. Rev. Lett., 104, 213905 (2010) - PDF
  • D Gachet, F Billard, H Rigneault, Focused field symmetries for background-free coherent anti-Stokes Raman spectroscopy, Phys. Rev. A 77, 0618021-4 (2008) - PDF
  • David Gachet, Franck Billard, Nicolas Sandeau, Herve Rigneault, Coherent anti-Stokes Raman scattering (CARS) microscopy imaging at interfaces : evidence of interference effects, Optics Express 15, 10408-10420 (2007) - PDF
  • N. Djaker, P.-F. Lenne, D. Marguet, A. Colonna, C. Hadjur, H. Rigneault
    Coherent Anti-Stokes Raman scattering microscopy (CARS) : Instrumentation and applications, Nuclear Instruments and Methods in Physics Research A Vol. 571, pp.177-181 (2007) - PDF