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High-contrast, synchronous volumetric imaging with selective volume illumination microscopy.

Thai V Truong | Daniel B Holland | Sara Madaan | Andrey Andreev | Kevin Keomanee-Dizon | Josh V Troll | Daniel E S Koo | Margaret J McFall-Ngai | Scott E Fraser
Communications biology | 2020

Light-field fluorescence microscopy uniquely provides fast, synchronous volumetric imaging by capturing an extended volume in one snapshot, but often suffers from low contrast due to the background signal generated by its wide-field illumination strategy. We implemented light-field-based selective volume illumination microscopy (SVIM), where illumination is confined to only the volume of interest, removing the background generated from the extraneous sample volume, and dramatically enhancing the image contrast. We demonstrate the capabilities of SVIM by capturing cellular-resolution 3D movies of flowing bacteria in seawater as they colonize their squid symbiotic partner, as well as of the beating heart and brain-wide neural activity in larval zebrafish. These applications demonstrate the breadth of imaging applications that we envision SVIM will enable, in capturing tissue-scale 3D dynamic biological systems at single-cell resolution, fast volumetric rates, and high contrast to reveal the underlying biology.

Pubmed ID: 32060411

Antibodies used in this publication

None found

Associated grants

  • Agency: NIMH NIH HHS, United States
    Id: R01 MH107238
  • Agency: NIH HHS, United States
    Id: R01 OD011024
  • Agency: NIAID NIH HHS, United States
    Id: R37 AI050661

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This is a list of tools and resources that we have found mentioned in this publication.


MATLAB (tool)

RRID:SCR_001622

Multi paradigm numerical computing environment and fourth generation programming language developed by MathWorks. Allows matrix manipulations, plotting of functions and data, implementation of algorithms, creation of user interfaces, and interfacing with programs written in other languages, including C, C++, Java, Fortran and Python. Used to explore and visualize ideas and collaborate across disciplines including signal and image processing, communications, control systems, and computational finance.

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Fiji (tool)

RRID:SCR_002285

Software package as distribution of ImageJ and ImageJ2 together with Java, Java3D and plugins organized into coherent menu structure. Used to assist research in life sciences.

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SPOT (tool)

RRID:SCR_005623

Software package for analysis of microarray images. Microarray spot detection and characterization software package which extracts numerical information from cDNA microarrays.

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Imaris (tool)

RRID:SCR_007370

Imaris provides range of capabilities for working with three dimensional images. Uses flexible editing and processing functions, such as interactive surface rendering and object slicing capabilities. And output to standard TIFF, Quicktime and AVI formats. Imaris accepts virtually all image formats that are used in confocal microscopy and many of those used in wide-field image acquisition.

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Zebrafish International Resource Center (biomaterial supply resource)

RRID:SCR_005065

Center that supplies access to wild-type, mutant, and transgenic zebrafish lines, EST's/cDNAs, antibodies and fish health services. ZIRC Health Services include diagnostic pathology testing for zebrafish and other small laboratory fish species.

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