Accueil > Français > Événements > Séminaires > Séminaires Matière Molle > Archives 2011

Séminaire de Willy Supatto

Fast and deep tissue imaging using nonlinear light sheet microscopy.

Willy Supatto

Laboratoire Optique et Biosciences, Ecole Polytechnique

Imaging fast beating cilia inside living zebrafish embryos or capturing cell movements deep inside highly scattering Drosophila tissues challenge the performances of current microscopy techniques. Fundamental light-matter interactions, such as light scattering, absorption, or photo-induced damage, set the limits of various imaging technologies in terms of spatial resolution, acquisition speed, and depth penetration. While standard two-photon excited fluorescence microscopy excels in achieving high depth penetration in scattering tissues, one-photon light sheet microscopy (SPIM, DSLM, etc) has gained widespread recognition in recent years, due to its distinct advantages for imaging live organisms with higher acquisition speed and lower phototoxicity. To find a new compromise point between these imaging performances, we report on the implementation and application of two-photon light sheet microscopy, combining two-photon excited fluorescence with the orthogonal illumination of light sheet microscopy. With live imaging of Drosophila and zebrafish embryos, we demonstrate the high performance of two-photon light sheet microscopy compared to the current state-of-the-art in maintaining good signal and high spatial resolution deep inside biological tissues, high acquisition speed and low phototoxicity. We further show the multi-modality of this approach by carrying out second harmonic generation light sheet microscopy to detect collagen in mouse tissue without fluorescent labeling.