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

Autor: Guillaume Labroille, Manuel Joffre, Israel Veilleux, Emmanuel Farge, Caroline Boudoux, Emmanuel Beaurepaire, Rajesh S. Pillai, Nicolas Olivier
Přispěvatelé: Laboratoire d'optique et biosciences (LOB), École polytechnique (X)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Ecole Polytechnique, École Polytechnique de Montréal (EPM)
Předmět:
Point spread function
Femtosecond pulse shaping
Fluorescence-lifetime imaging microscopy
Optics and Photonics
Materials science
Embryo
Nonmammalian
MESH: Drosophila
[SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging
MESH: Developmental Biology
Green Fluorescent Proteins
Physics::Optics
02 engineering and technology
MESH: Signal Processing
Computer-Assisted
01 natural sciences
010309 optics
Optics
MESH: Green Fluorescent Proteins
Two-photon excitation microscopy
0103 physical sciences
Microscopy
Image Processing
Computer-Assisted
Animals
MESH: Animals
[PHYS.PHYS.PHYS-OPTICS]Physics [physics]/Physics [physics]/Optics [physics.optics]
Spatial light modulator
Models
Statistical
Fourier Analysis
business.industry
Lasers
MESH: Embryo
Nonmammalian
Signal Processing
Computer-Assisted
Equipment Design
021001 nanoscience & nanotechnology
Pulse shaping
MESH: Image Processing
Computer-Assisted
Atomic and Molecular Physics
and Optics
Microscopy
Fluorescence
Multiphoton
Coherent control
MESH: Microscopy
Fluorescence
Multiphoton
MESH: Lasers
Drosophila
0210 nano-technology
business
MESH: Optics and Photonics
MESH: Models
Statistical
MESH: Fourier Analysis
Developmental Biology
MESH: Equipment Design
Zdroj: ResearcherID
Optics Express
Optics Express, Optical Society of America-OSA Publishing, 2009, 17 (15), pp.12741-52. ⟨10.1364/OE.17.012741⟩
ISSN: 1094-4087
DOI: 10.1364/OE.17.012741⟩
Popis: International audience; 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.
Databáze: OpenAIRE
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