Three-dimensional wide-field fluorescence microscopy for transcranial mapping of cortical microcirculation
Wide-field fluorescence imaging is an indispensable tool for studying largescale biodynamics. Limited space-bandwidth product and strong light diffusion make conventional implementations incapable of high-resolution mapping of fluorescence biodistribution in three dimensions. We introduce a volumetric wide-field fluorescence microscopy based on optical astigmatism combined with fluorescence source localization, covering 5.6×5.6×0.6 mm3 imaging volume. Two alternative configurations are proposed exploitingmultifocal illumination or sparse localization of point emitters, which are herein seamlessly integrated in one system.We demonstrate real-time volumetricmapping of the murine cortical microcirculation at capillary resolution without employing cranial windows, thus simultaneously delivering quantitative perfusion information across both brain hemispheres. Morphological and functional changes of cerebral vascular networks are further investigated after an acute ischemic stroke, enabling cortex-wide observation of concurrent collateral recruitment events occurring on a sub-second scale. The reported technique thus offers a wealth of unmatched possibilities for non- orminimally invasive imaging of biodynamics across scales.
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