Multicolor two-photon tissue imaging by wavelength mixing.

Mahou, Pierre; Zimmerley, Maxwell; Loulier, Karine; Matho, Katherine S; Labroille, Guillaume; Morin, Xavier; Supatto, Willy; Livet, Jean et al. · Nat Methods · 2012

basic_science · Level V

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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.

Medical subject headings