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

Rajesh Pillai 1 Caroline Boudoux 1, 2 Guillaume Labroille 1 Nicolas Olivier 1 Israel Veilleux 1 Emmanuel Farge 2 Manuel Joffre 1 Emmanuel Beaurepaire 1, *
* Corresponding author
1 LOB - Laboratoire d'optique et biosciences
2 Ecole Polytechnique
Abstract : 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.
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Rajesh 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, Optical Society of America, 2009, 17 (15), pp.12741-52. ⟨10.1364/OE.17.012741⟩. ⟨hal-00818488⟩

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