biostudies-literatureLi HNational Science Foundation (NSF)NIGMS NIH HHSNational Institute of General Medical Sciences (NIGMS)29-49https://www.ebi.ac.uk/biostudies/studies/S-EPMC6363205biostudies-literatureUnknown10(1)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.Biomedical optics expressFast, volumetric live-cell imaging using high-resolution light-field microscopy.PMC6363205R01GM084251R35 GM124898R35GM124846R01 GM084251R35 GM124846EFMA1830941CBET1604565Li WTakamaru KILi HAltshuller YJia SFrench JBGuo CKim-Holzapfel DMeng YFrohman MASchroeder BLiu WfalseFast, volumetric live-cell imaging using high-resolution light-field microscopy.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.2019-01-01T00:00:00Z2019 Jan2026-07-11T03:18:15.31Z2026-07-11T03:09:47.043ZS-EPMC63632053077508110.1364/BOE.10.00002910.1364/boe.10.000029