Multicolor two-photon tissue imaging by wavelength mixing

Pierre Mahou (1) , Maxwell Zimmerley (1) , Karine Loulier (2) , Katherine S. Matho (2) , Guillaume Labroille (1) , Xavier Morin (3) , Willy Supatto (1) , Jean Livet (2) , Delphine Débarre (1) , Emmanuel Beaurepaire (1)

1 LOB - Laboratoire d'optique et biosciences
2 Institut de la Vision
3 IBENS - Institut de biologie de l'ENS Paris

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.

Domaines: Sciences du Vivant [q-bio]

Pierre Mahou, Maxwell Zimmerley, Karine Loulier, Katherine S. Matho, Guillaume Labroille, et al.. Multicolor two-photon tissue imaging by wavelength mixing. Nature Methods, 2012, 9 (8), pp.815-818. ⟨10.1038/nmeth.2098⟩. ⟨hal-00324345⟩