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Multiphoton microscopy of developing tissues

PIs & engineers: Emmanuel Beaurepaire, Willy Supatto, Chiara Stringari, Pierre Mahou, Marie-Claire Schanne-Klein

 

Our group associates expertise in nonlinear optics and tissue microscopy. We develop novel experimental approaches based on multimodal multiphoton microscopy (*), and their application to the study of developing tissues.

Current issues in systems biology (development, neuroscience) require tissue-scale measurements of multiple cell parameters. Since its introduction in the early 90s, multiphoton microscopy has proven invaluable for tissue studies with its ability to provide micrometer-scale 3D resolution in intact, scattering samples. However tissue imaging is still challenging in terms of speed, depth, sensitivity, and number of accessible parameters.
We develop strategies to expand the capabilities of current microscopes (characterization of coherent contrast mechanisms; multimodal/multicolor imaging of several signals; light-sheet excitation for faster imaging; wavefront control for focus engineering and adaptive optics). In relation with the other projects of the 'advanced microscopies' pole, we explore novel applications such as imaging embryonic & nervous tissue development in animal models, and optical diagnostics.

See also:
   - Multicolor two-photon microscopy (wavelength mixing, brainbow)
   - Two-photon light sheet microscopy (2P-SPIM)
   - FLIM-based metabolic imaging
   - Deep-tissue 3-photon (3PEF) microscopy
   - Label-free third harmonic generation (THG) microscopy
   - Wavefront control (adaptive optics, Bessel beams)

(*) Combining photomanipulation and multiphoton contrasts such as multiphoton-excited fluorescence (2PEF, 3PEF), harmonic generation (THG, SHG), coherent anti-Stokes Raman scattering (CARS), four-wave mixing (FWM), fluorescence lifetime (FLIM), polarimetry.


Some introduction articles

(Review article) "Advances in whole-embryo imaging: a quantitative transition is underway", Pantazis & Supatto, Nat Rev Mol Cell Biol (2014).
(French) "Imagerie du vivant: des couleurs pour étudier la complexité", Beaurepaire & Livet, Huffington Post (2013).
(French) "Microscopie multiphoton illuminée par nappe : imagerie de fluorescence rapide et en profondeur dans les tissus vivants", Supatto, Photoniques (2012).
(Review article) "Advances in multiphoton microscopy for imaging embryos" Supatto et al, Curr Opin Gen Dev (2011).
(French video) Reconstruction du lignage cellulaire de l'embryon du poisson-zébré (2010).
(English video) Cell lineage reconstruction of early zebrafish embryos (2010).
(French) "Microscopies multi-harmoniques pour l'imagerie structurale de tissus intacts", Débarre et al. Medecine/Sciences (2006).

THG-SHG imaging of a live zebrafish embryo. SHG (green) reveals the mitotic spindles in dividing cells. THG (blue) shows the morphology of the cells and vitellus. (Olivier et al, Science 339, 967(2010))