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. 2012 Jul 8;9(8):815-8.
doi: 10.1038/nmeth.2098.

Multicolor two-photon tissue imaging by wavelength mixing

Pierre Mahou  1 , Maxwell Zimmerley, Karine Loulier, Katherine S Matho, Guillaume Labroille, Xavier Morin, Willy Supatto, Jean Livet, Delphine Débarre, Emmanuel Beaurepaire
Affiliations

Multicolor two-photon tissue imaging by wavelength mixing

Pierre Mahou et al. Nat Methods. .

Abstract

We achieve simultaneous two-photon excitation of three chromophores with distinct absorption spectra using synchronized pulses from a femtosecond laser and an optical parametric oscillator. The two beams generate separate multiphoton processes, and their spatiotemporal overlap provides an additional two-photon excitation route, with submicrometer overlay of the color channels. We report volume and live multicolor imaging of 'Brainbow'-labeled tissues as well as simultaneous three-color fluorescence and third-harmonic imaging of fly embryos.

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