Observing the Cell in Its Native State: Imaging Subcellular Dynamics in Multicellular Organisms

Author:

Liu Tsung-LiORCID,Upadhyayula SrigokulORCID,Milkie Daniel E.,Singh Ved,Wang Kai,Swinburne Ian A.,Mosaliganti Kishore R.,Collins Zach M.ORCID,Hiscock Tom W.ORCID,Shea Jamien,Kohrman Abraham Q.ORCID,Medwig Taylor N.,Dambournet Daphne,Forster Ryan,Cunniff BrianORCID,Ruan Yuan,Yashiro Hanako,Scholpp SteffenORCID,Meyerowitz Elliot M.ORCID,Hockemeyer Dirk,Drubin David G.ORCID,Martin Benjamin L.,Matus David Q.,Koyama Minoru,Megason Sean G.ORCID,Kirchhausen Tom,Betzig EricORCID

Abstract

AbstractTrue physiological imaging of subcellular dynamics requires studying cells within their parent organisms, where all the environmental cues that drive gene expression, and hence the phenotypes we actually observe, are present. A complete understanding also requires volumetric imaging of the cell and its surroundings at high spatiotemporal resolution without inducing undue stress on either. We combined lattice light sheet microscopy with two-channel adaptive optics to achieve, across large multicellular volumes, noninvasive aberration-free imaging of subcellular processes, including endocytosis, organelle remodeling during mitosis, and the migration of axons, immune cells, and metastatic cancer cells in vivo. The technology reveals the phenotypic diversity within cells across different organisms and developmental stages, and may offer insights into how cells harness their intrinsic variability to adapt to different physiological environments.One Sentence SummaryCombining lattice light sheet microscopy with adaptive optics enables high speed, high resolution in vivo 3D imaging of dynamic processes inside cells under physiological conditions within their parent organisms.

Publisher

Cold Spring Harbor Laboratory

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