
Simultaneous multiview capture and fusion improves spatial resolution in wide-field and light-sheet microscopy
Author(s) -
Yicong Wu,
Panagiotis Chandris,
Peter W. Winter,
Edward Y. Kim,
Valentin Jaumouillé,
Abhishek Kumar,
Min Guo,
Jacqueline M. Leung,
Claire Smith,
Ivan Rey-Suarez,
Huafeng Liu,
Clare M. Waterman,
Kumaran S. Ramamurthi,
Patrick J. La Rivière,
Hari Shroff
Publication year - 2016
Publication title -
optica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.074
H-Index - 107
ISSN - 2334-2536
DOI - 10.1364/optica.3.000897
Subject(s) - light sheet fluorescence microscopy , microscopy , image resolution , resolution (logic) , optics , microscope , deconvolution , materials science , computer science , biological system , computer vision , artificial intelligence , physics , scanning confocal electron microscopy , biology
Most fluorescence microscopes are inefficient, collecting only a small fraction of the emitted light at any instant. Besides wasting valuable signal, this inefficiency also reduces spatial resolution and causes imaging volumes to exhibit significant resolution anisotropy. We describe microscopic and computational techniques that address these problems by simultaneously capturing and subsequently fusing and deconvolving multiple specimen views. Unlike previous methods that serially capture multiple views, our approach improves spatial resolution without introducing any additional illumination dose or compromising temporal resolution relative to conventional imaging. When applying our methods to single-view wide-field or dual-view light-sheet microscopy, we achieve a twofold improvement in volumetric resolution (~235 nm × 235 nm × 340 nm) as demonstrated on a variety of samples including microtubules in Toxoplasma gondii , SpoVM in sporulating Bacillus subtilis , and multiple protein distributions and organelles in eukaryotic cells. In every case, spatial resolution is improved with no drawback by harnessing previously unused fluorescence.