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Reflective imaging improves spatiotemporal resolution and collection efficiency in light sheet microscopy

Author

Listed:
  • Yicong Wu

    (National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health)

  • Abhishek Kumar

    (National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health)

  • Corey Smith

    (University of Chicago)

  • Evan Ardiel

    (National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health)

  • Panagiotis Chandris

    (National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health)

  • Ryan Christensen

    (National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health)

  • Ivan Rey-Suarez

    (National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health
    University of Maryland)

  • Min Guo

    (National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health)

  • Harshad D. Vishwasrao

    (Advanced Imaging and Microscopy Resource, National Institutes of Health)

  • Jiji Chen

    (Advanced Imaging and Microscopy Resource, National Institutes of Health)

  • Jianyong Tang

    (JT Scientific Consulting LLC)

  • Arpita Upadhyaya

    (University of Maryland
    University of Maryland)

  • Patrick J. La Riviere

    (University of Chicago
    Marine Biological Laboratory)

  • Hari Shroff

    (National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health
    Advanced Imaging and Microscopy Resource, National Institutes of Health
    University of Maryland
    Marine Biological Laboratory)

Abstract

Light-sheet fluorescence microscopy (LSFM) enables high-speed, high-resolution, and gentle imaging of live specimens over extended periods. Here we describe a technique that improves the spatiotemporal resolution and collection efficiency of LSFM without modifying the underlying microscope. By imaging samples on reflective coverslips, we enable simultaneous collection of four complementary views in 250 ms, doubling speed and improving information content relative to symmetric dual-view LSFM. We also report a modified deconvolution algorithm that removes associated epifluorescence contamination and fuses all views for resolution recovery. Furthermore, we enhance spatial resolution (to

Suggested Citation

  • Yicong Wu & Abhishek Kumar & Corey Smith & Evan Ardiel & Panagiotis Chandris & Ryan Christensen & Ivan Rey-Suarez & Min Guo & Harshad D. Vishwasrao & Jiji Chen & Jianyong Tang & Arpita Upadhyaya & Pat, 2017. "Reflective imaging improves spatiotemporal resolution and collection efficiency in light sheet microscopy," Nature Communications, Nature, vol. 8(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_s41467-017-01250-8
    DOI: 10.1038/s41467-017-01250-8
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