Multiview confocal super-resolution microscopy.

Journal: Nature
PMID: 34837071

Abstract

Confocal microscopy remains a major workhorse in biomedical optical microscopy owing to its reliability and flexibility in imaging various samples, but suffers from substantial point spread function anisotropy, diffraction-limited resolution, depth-dependent degradation in scattering samples and volumetric bleaching. Here we address these problems, enhancing confocal microscopy performance from the sub-micrometre to millimetre spatial scale and the millisecond to hour temporal scale, improving both lateral and axial resolution more than twofold while simultaneously reducing phototoxicity. We achieve these gains using an integrated, four-pronged approach: (1) developing compact line scanners that enable sensitive, rapid, diffraction-limited imaging over large areas; (2) combining line-scanning with multiview imaging, developing reconstruction algorithms that improve resolution isotropy and recover signal otherwise lost to scattering; (3) adapting techniques from structured illumination microscopy, achieving super-resolution imaging in densely labelled, thick samples; (4) synergizing deep learning with these advances, further improving imaging speed, resolution and duration. We demonstrate these capabilities on more than 20 distinct fixed and live samples, including protein distributions in single cells; nuclei and developing neurons in Caenorhabditis elegans embryos, larvae and adults; myoblasts in imaginal disks of Drosophila wings; and mouse renal, oesophageal, cardiac and brain tissues.

Authors

  • Yicong Wu
    Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA. yicong.wu@nih.gov.
  • Xiaofei Han
    Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA.
  • Yijun Su
    Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA.
  • Melissa Glidewell
    Applied Scientific Instrumentation, Eugene, OR, USA.
  • Jonathan S Daniels
    Applied Scientific Instrumentation, Eugene, OR, USA.
  • Jiamin Liu
  • Titas Sengupta
    Department of Neuroscience and Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA.
  • Ivan Rey-Suarez
    Institute for Physical Science and Technology, University of Maryland, College Park, MD, USA.
  • Robert Fischer
    Cell and Developmental Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
  • Akshay Patel
    Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, USA.
  • Christian Combs
    NHLBI Light Microscopy Facility, National Institutes of Health, Bethesda, MD, USA.
  • Junhui Sun
    Laboratory of Cardiac Physiology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
  • Xufeng Wu
    NHLBI Light Microscopy Facility, National Institutes of Health, Bethesda, MD, USA.
  • Ryan Christensen
    Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA.
  • Corey Smith
    Department of Mechanical and Materials Engineering, The University of Western Ontario, London, Ontario, Canada.
  • Lingyu Bao
    Section on Molecular Morphogenesis, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD, USA.
  • Yilun Sun
    Department of Radiation Oncology, University Hospitals Seidman Cancer Center, Case Western Reserve University, Cleveland.
  • Leighton H Duncan
    Department of Neuroscience and Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA.
  • Jiji Chen
    Advanced Imaging and Microscopy Resource, National Institutes of Health, Bethesda, MD, USA.
  • Yves Pommier
    Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, Center for Cancer Research, National Institutes of Health, Bethesda, MD, USA.
  • Yun-Bo Shi
    Section on Molecular Morphogenesis, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD, USA.
  • Elizabeth Murphy
    Laboratory of Cardiac Physiology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
  • Sougata Roy
    Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, USA.
  • Arpita Upadhyaya
    Institute for Physical Science and Technology, University of Maryland, College Park, MD, USA.
  • Daniel Colón-Ramos
    Department of Neuroscience and Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA.
  • Patrick La Riviere
    Department of Radiology, University of Chicago, Chicago, IL, USA.
  • Hari Shroff
    Laboratory of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA.

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