Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution

单物镜光片显微镜可实现高速全细胞 3D 超分辨率

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作者:Marjolein B M Meddens, Sheng Liu, Patrick S Finnegan, Thayne L Edwards, Conrad D James, Keith A Lidke

Abstract

We have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single molecule super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet.

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