Open AccessSingle objective light-sheet microscopy for high-speed whole-cell 3D super-resolution
Author(s) -
Marjolein B.M. Meddens,
Sheng Liu,
Patrick Sean Finnegan,
Thayne L. Edwards,
Conrad D. James,
Keith A. Lidke
Publication year - 2016
Publication title -
biomedical optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.7.002219
Subject(s) - light sheet fluorescence microscopy , optics , numerical aperture , microscopy , materials science , lens (geology) , image quality , optical microscope , resolution (logic) , bright field microscopy , optical sectioning , microscope , focus (optics) , aperture (computer memory) , light field , cardinal point , optoelectronics , physics , scanning confocal electron microscopy , computer science , scanning electron microscope , computer vision , artificial intelligence , wavelength , acoustics , image (mathematics)
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.