Séminaire d'Alexandra Fragola, en mode HYBRIDE

Structured illumination and adaptive optics fluorescence microscopy for high resolution bio-imaging

par Alexandra Fragola

ISMO, Université Paris-Saclay/CNRS

Fluorescence microscopy is a technique of choice for dynamic imaging of living samples. However, image quality remains limited by diffraction or optical aberrations, which restrict both resolution and sensitivity in all microscopy modalities. To overcome these difficulties, we developed new optical microscopes for high-resolution imaging of living biological samples, from the individual cell to the small animal. Structured illumination microscopy (SIM) allows to exceed the diffraction limit with only a few acquired images, thus allowing dynamic super resolution imaging of living cells. As shown in figure 1a, the resolution gain of SIM up to 100nm allows to study the dynamics of the mitochondrial cristae in cells subjected to a photoinduced stress. For in depth imaging inside biological tissues, adaptive optics provides a reliable live correction of the aberrations and thus a significant gain in both spatial resolution and signal intensity. We proposed a few years ago an innovative strategy of adaptive optics, by adapting pioneer work from astronomy to the constraints of fluorescence light-sheet microscopy and two-photon microscopy, and demonstrated its efficiency for enhanced live imaging of Zebrafish larvae and structural or functional neuroimaging of ex vivo Drosophila brains and mouse brain slices (see figure 1b).
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