Multiplexed two-photon microscopy of dynamic biological samples with shaped broadband pulses.

Rajesh S. Pillai (1) , Caroline Boudoux (1, 2) , Guillaume Labroille (1) , Nicolas Olivier (1) , Israel Veilleux (1) , Emmanuel Farge (2) , Manuel Joffre (1) , Emmanuel Beaurepaire (1)

1 LOB - Laboratoire d'optique et biosciences
2 Ecole Polytechnique

Coherent control can be used to selectively enhance or cancel concurrent multiphoton processes, and has been suggested as a means to achieve nonlinear microscopy of multiple signals. Here we report multiplexed two-photon imaging in vivo with fast pixel rates and micrometer resolution. We control broadband laser pulses with a shaping scheme combining diffraction on an optically-addressed spatial light modulator and a scanning mirror allowing to switch between programmable shapes at kiloHertz rates. Using coherent control of the two-photon excited fluorescence, it was possible to perform selective microscopy of GFP and endogenous fluorescence in developing Drosophila embryos. This study establishes that broadband pulse shaping is a viable means for achieving multiplexed nonlinear imaging of biological tissues.

Domains: Physics [physics]; Physics [physics]; Optics [physics.optics]; Life Sciences [q-bio]; Bioengineering; Imaging

Rajesh S. Pillai, Caroline Boudoux, Guillaume Labroille, Nicolas Olivier, Israel Veilleux, et al.. Multiplexed two-photon microscopy of dynamic biological samples with shaped broadband pulses.. Optics Express, 2009, 17 (15), pp.12741-52. ⟨10.1364/OE.17.012741⟩. ⟨hal-00818488⟩