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. Show more
Permanent link
https://doi.org/10.3929/ethz-b-000387371Publication status
publishedExternal links
Journal / series
Laser & Photonics ReviewsVolume
Pages / Article No.
Publisher
WileySubject
Diffraction gratings; Fast scanning microscopy; Fluorescence imaging; Multifocal illuminationOrganisational unit
09648 - Razansky, Daniel / Razansky, Daniel
09648 - Razansky, Daniel / Razansky, Daniel
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