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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 2, Iss. 10 — Oct. 1, 2011
  • pp: 2837–2849

Combined third-harmonic generation and four-wave mixing microscopy of tissues and embryos

Pierre Mahou, Nicolas Olivier, Guillaume Labroille, Louise Duloquin, Jean-Marc Sintes, Nadine Peyriéras, Renaud Legouis, Delphine Débarre, and Emmanuel Beaurepaire  »View Author Affiliations

Biomedical Optics Express, Vol. 2, Issue 10, pp. 2837-2849 (2011)

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Nonlinear microscopy can be used to probe the intrinsic optical properties of biological tissues. Using femtosecond pulses, third-harmonic generation (THG) and four-wave mixing (FWM) signals can be efficiently produced and detected simultaneously. Both signals probe a similar parameter, i.e. the real part of the third-order nonlinear susceptibility χ(3). However THG and FWM images result from different phase matching conditions and provide complementary information. We analyze this complementarity using calculations, z-scan measurements on water and oils, and THG-FWM imaging of cell divisions in live zebrafish embryos. The two signals exhibit different sensitivity to sample size and clustering in the half-wavelength regime. Far from resonance, THG images reveal spatial variations |Δχ(3)(−3ω;ω,ω,ω)| with remarkable sensitivity while FWM directly reflects the distribution of χ(3)(−2ω1 + ω2;ω1, –ω2, ω1). We show that FWM images provide χ(3) maps useful for proper interpretation of cellular THG signals, and that combined imaging carries additional structural information. Finally we present simultaneous imaging of intrinsic THG, FWM, second-harmonic (SHG) and two-photon-excited fluorescence (2PEF) signals in live Caenorhabditis elegans worms illustrating the information provided by multimodal nonlinear imaging of unstained tissue.

© 2011 OSA

OCIS Codes
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(190.4160) Nonlinear optics : Multiharmonic generation
(190.4380) Nonlinear optics : Nonlinear optics, four-wave mixing
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:

Original Manuscript: August 19, 2011
Revised Manuscript: September 19, 2011
Manuscript Accepted: September 19, 2011
Published: September 26, 2011

Pierre Mahou, Nicolas Olivier, Guillaume Labroille, Louise Duloquin, Jean-Marc Sintes, Nadine Peyriéras, Renaud Legouis, Delphine Débarre, and Emmanuel Beaurepaire, "Combined third-harmonic generation and four-wave mixing microscopy of tissues and embryos," Biomed. Opt. Express 2, 2837-2849 (2011)

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