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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editor: Gregory W. Faris
  • Vol. 4, Iss. 9 — Sep. 4, 2009

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

Rajesh S. Pillai, Caroline Boudoux, Guillaume Labroille, Nicolas Olivier, Israel Veilleux, Emmanuel Farge, Manuel Joffre, and Emmanuel Beaurepaire  »View Author Affiliations

Optics Express, Vol. 17, Issue 15, pp. 12741-12752 (2009)

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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.

© 2009 Optical Society of America

OCIS Codes
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(180.6900) Microscopy : Three-dimensional microscopy
(230.6120) Optical devices : Spatial light modulators
(320.5540) Ultrafast optics : Pulse shaping
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: March 17, 2009
Revised Manuscript: May 4, 2009
Manuscript Accepted: May 6, 2009
Published: July 13, 2009

Virtual Issues
Vol. 4, Iss. 9 Virtual Journal for Biomedical Optics

Rajesh S. Pillai, Caroline Boudoux, Guillaume Labroille, Nicolas Olivier, Israel Veilleux, Emmanuel Farge, Manuel Joffre, and Emmanuel Beaurepaire, "Multiplexed two-photon microscopy of dynamic biological samples with shaped broadband pulses," Opt. Express 17, 12741-12752 (2009)

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