High-Speed Large-Field Multifocal Illumination Fluorescence Microscopy

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Published on:
Authors: Chen, Zhenyue; Mc Larney, Benedict; Rebling, Johannes; Deán-Ben, Xosé Luis; Zhou, Quanyu; Gottschalk, Sven & Razansky, Daniel
Abstract:

“Abstract Scanning optical microscopy techniques are commonly restricted to a sub-millimeter field-of-view (FOV) or otherwise employ slow mechanical translation, limiting their applicability for imaging fast biological dynamics occurring over large areas. A rapid scanning large-field multifocal illumination (LMI) fluorescence microscopy technique is devised based on a beam-splitting grating and an acousto-optic deflector synchronized with a high-speed camera to attain real-time fluorescence microscopy over a centimeter-scale FOV. Owing to its large depth of focus, the approach allows noninvasive visualization of perfusion across the entire mouse cerebral cortex, not achievable with conventional wide-field fluorescence microscopy methods. The new concept can readily be incorporated into conventional wide-field microscopes to mitigate image blur due to tissue scattering and attain optimal trade-off between spatial resolution and FOV. It further establishes a bridge between conventional wide-field macroscopy and laser scanning confocal microscopy, thus it is anticipated to find broad applicability in functional neuroimaging, in vivo cell tracking, and other applications looking at large-scale fluorescent-based biodynamics.”

Open Access

Publication: Laser & Photonics Reviews
Issue/Year: Laser & Photonics Reviews, Volume n/a; Number n/a; Pages 1900070; 2019
DOI: 10.1002/lpor.201900070
Link: https://doi.org/10.1002/lpor.201900070

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