title: Simultaneous multiview capture and fusion improves spatial resolution in wide-field and light-sheet microscopy.
abstract: 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 inToxoplasma gondii, SpoVM in sporulatingBacillus 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.
authors.affiliation: Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Yicong
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Chandris
authors.affiliation: Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Peter W
authors.initials: PW
authors.lastname: Winter
authors.affiliation: Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Edward Y
authors.initials: EY
authors.lastname: Kim
authors.affiliation: Cell Biology and Physiology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Valentin
authors.initials: V
authors.lastname: Jaumouillé
authors.affiliation: Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland 20892, USA; State Key Lab of Modern Optical Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Min
authors.initials: M
authors.lastname: Guo
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, Indiana 47405, USA.
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Leung
authors.affiliation: Department of Radiology, University of Chicago, Chicago, Illinois 60637, USA.
authors.forename: Corey
authors.initials: C
authors.lastname: Smith
authors.affiliation: Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Ivan
authors.initials: I
authors.lastname: Rey-Suarez
authors.affiliation: State Key Lab of Modern Optical Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Huafeng
authors.initials: H
authors.lastname: Liu
authors.affiliation: Cell Biology and Physiology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Clare M
authors.initials: CM
authors.lastname: Waterman
authors.affiliation: Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Kumaran S
authors.initials: KS
authors.lastname: Ramamurthi
authors.affiliation: Department of Radiology, University of Chicago, Chicago, Illinois 60637, USA; Whitman Center, Marine Biological Laboratory, Woods Hole, Massachusetts 02543, USA.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: La Riviere
authors.affiliation: Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland 20892, USA; Whitman Center, Marine Biological Laboratory, Woods Hole, Massachusetts 02543, USA.
authors.forename: Hari
authors.initials: H
authors.lastname: Shroff