Fast, volumetric live-cell imaging using high-resolution light-field microscopy

• Type of Material: Article
• Authors: Haoyu Li, SUNY Stony Brook; Changliang Guo, Georgia Institute of Technology; Deborah Kim-Holzapfel, SUNY Stony Brook; Weiyi Li, SUNY Stony Brook; Yelena Altshuller, SUNY Stony Brook; Bryce Schroeder, SUNY Stony Brook; Wenhao Liu, Georgia Institute of Technology; Yizhi Meng, SUNY Stony Brook; Jarrod B. French, SUNY Stony Brook; Ken-Ichi Takamaru, SUNY Stony Brook; Michael A. Frohman, SUNY Stony Brook; Shu Jia, Emory University
• Language: English
• Date: 2019-01-01
• Publisher: Optical Society of America: Open Access Journals
• Publication Version: Final Published Version
• Copyright Statement: © 2018 Optical Society of America.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1364/BOE.10.000029
• Title of Journal or Parent Work: Biomedical Optics Express
• ISSN: 2156-7085
• Volume: 10
• Issue: 1
• Start Page: 29
• End Page: 49
• Grant/Funding Information: National Institutes of Health grants 1R35GM124846; (to S.J.) and R01GM084251 (to M.F.), National Science Foundation grants CBET1604565; and EFMA1830941 (to S.J.).; We acknowledge the support of the NSF-CBET Biophotonics program, the NSF-EFMA program, and the NIH-NIGMS MIRA program.
• Keywords: Biochemical Research Methods; NEURONAL-ACTIVITY; Biochemistry & Molecular Biology; Radiology, Nuclear Medicine & Medical Imaging; Life Sciences & Biomedicine; Science & Technology; Optics; Physical Sciences
• Research Categories: Chemistry, Biochemistry; Health Sciences, Radiology; Chemistry, Physical

Abstract

Visualizing diverse anatomical and functional traits that span many spatial scales with high spatio-temporal resolution provides insights into the fundamentals of living organisms. Light-field microscopy (LFM) has recently emerged as a scanning-free, scalable method that allows for high-speed, volumetric functional brain imaging. Given those promising applications at the tissue level, at its other extreme, this highly-scalable approach holds great potential for observing structures and dynamics in single-cell specimens. However, the challenge remains for current LFM to achieve a subcellular level, near-diffraction-limited 3D spatial resolution. Here, we report high-resolution LFM (HR-LFM) for live-cell imaging with a resolution of 300-700 nm in all three dimensions, an imaging depth of several micrometers, and a volume acquisition time of milliseconds. We demonstrate the technique by imaging various cellular dynamics and structures and tracking single particles. The method may advance LFM as a particularly useful tool for understanding biological systems at multiple spatio-temporal levels.