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

• Published in: Communications biology Vol. 3; no. 1; p. 74
• Main Authors: Truong, Thai V., Holland, Daniel B., Madaan, Sara, Andreev, Andrey, Keomanee-Dizon, Kevin, Troll, Josh V., Koo, Daniel E. S., McFall-Ngai, Margaret J., Fraser, Scott E.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 14.02.2020; Nature Publishing Group
• Subjects: 14; 14/63; 14/69; 3-D films; 631/1647/328/2236; 631/1647/328/2237; Animals; Bacteria; Biology; Biomedical and Life Sciences; Brain - anatomy & histology; Brain - diagnostic imaging; Brain - ultrastructure; Danio rerio; Decapodiformes - microbiology; Decapodiformes - ultrastructure; Fluorescence microscopy; Heart - anatomy & histology; Heart - diagnostic imaging; Heart - physiology; Host Microbial Interactions - physiology; Illumination; Image Processing, Computer-Assisted - instrumentation; Image Processing, Computer-Assisted - methods; Imaging, Three-Dimensional - instrumentation; Imaging, Three-Dimensional - methods; Larva; Life Sciences; Light; Microscopy; Microscopy, Fluorescence - instrumentation; Microscopy, Fluorescence - methods; Neuroimaging; Organ Size; Seawater - microbiology; Video Recording - instrumentation; Video Recording - methods; Zebrafish
• ISSN: 2399-3642; 2399-3642
• DOI: 10.1038/s42003-020-0787-6

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. Thai Truong et al. present light-field-based selective volume illumination microscopy (SVIM), a method for enhancing image contrast and resolution by combining light-field microscopy and selective plane illumination microscopy. They generate cellular-resolution 3D movies by applying SVIM to flowing bacteria in seawater and to the beating heart and whole brain of larval zebrafish.