title: Lattice light-sheet microscopy: imaging molecules to embryos at high spatiotemporal resolution.
abstract: Although fluorescence microscopy provides a crucial window into the physiology of living specimens, many biological processes are too fragile, are too small, or occur too rapidly to see clearly with existing tools. We crafted ultrathin light sheets from two-dimensional optical lattices that allowed us to image three-dimensional (3D) dynamics for hundreds of volumes, often at subsecond intervals, at the diffraction limit and beyond. We applied this to systems spanning four orders of magnitude in space and time, including the diffusion of single transcription factor molecules in stem cell spheroids, the dynamic instability of mitotic microtubules, the immunological synapse, neutrophil motility in a 3D matrix, and embryogenesis in Caenorhabditis elegans and Drosophila melanogaster. The results provide a visceral reminder of the beauty and the complexity of living systems.
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Bi-Chang
authors.initials: BC
authors.lastname: Chen
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Wesley R
authors.initials: WR
authors.lastname: Legant
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Kai
authors.initials: K
authors.lastname: Wang
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Lin
authors.initials: L
authors.lastname: Shao
authors.affiliation: Coleman Technologies, Incorporated, Newtown Square, PA 19073, USA.
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Milkie
authors.affiliation: National High Magnetic Field Laboratory and Department of Biological Science, Florida State University, Tallahassee, FL 32310, USA.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Davidson
authors.affiliation: Department of Biological Sciences, University of the Sciences, Philadelphia, PA 19104, USA.
authors.forename: Chris
authors.initials: C
authors.lastname: Janetopoulos
authors.affiliation: Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Xufeng S
authors.initials: XS
authors.lastname: Wu
authors.affiliation: Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Hammer
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
authors.forename: Brian P
authors.initials: BP
authors.lastname: English
authors.affiliation: Optical Image Analysis Unit, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Mimori-Kiyosue
authors.affiliation: Department of Pharmacology, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Romero
authors.affiliation: Cell Biology and Metabolism Program, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. Cambridge Institute for Medical Research, Addenbrooke's Hospital, Cambridge CB2 0XY, England, UK.
authors.forename: Alex T
authors.initials: AT
authors.lastname: Ritter
authors.affiliation: Cell Biology and Metabolism Program, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Lippincott-Schwartz
authors.affiliation: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: Lillian
authors.initials: L
authors.lastname: Fritz-Laylin
authors.affiliation: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: R Dyche
authors.initials: RD
authors.lastname: Mullins
authors.affiliation: Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA.
authors.forename: Diana M
authors.initials: DM
authors.lastname: Mitchell
authors.affiliation: Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA.
authors.forename: Joshua N
authors.initials: JN
authors.lastname: Bembenek
authors.affiliation: Max Planck Institute for Molecular Cell Biology and Genetics, 01307 Dresden, Germany. Max Planck Institute for the Physics of Complex Systems, 01307 Dresden, Germany.
authors.forename: Anne-Cecile
authors.initials: AC
authors.lastname: Reymann
authors.affiliation: Max Planck Institute for Molecular Cell Biology and Genetics, 01307 Dresden, Germany. Max Planck Institute for the Physics of Complex Systems, 01307 Dresden, Germany.
authors.forename: Ralph
authors.initials: R
authors.lastname: Böhme
authors.affiliation: Max Planck Institute for Molecular Cell Biology and Genetics, 01307 Dresden, Germany. Max Planck Institute for the Physics of Complex Systems, 01307 Dresden, Germany.
authors.forename: Stephan W
authors.initials: SW
authors.lastname: Grill
authors.affiliation: Department of Molecular Biology and Genetics, Howard Hughes Medical Institute, Center for Cell Dynamics, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Jennifer T
authors.initials: JT
authors.lastname: Wang
authors.affiliation: Department of Molecular Biology and Genetics, Howard Hughes Medical Institute, Center for Cell Dynamics, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Geraldine
authors.initials: G
authors.lastname: Seydoux
authors.affiliation: Department of Biology, Duke University, Durham, NC 27708, USA.
authors.forename: U Serdar
authors.initials: US
authors.lastname: Tulu
authors.affiliation: Department of Biology, Duke University, Durham, NC 27708, USA.
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Kiehart
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA. betzige@janelia.hhmi.org.
authors.forename: Eric
authors.initials: E
authors.lastname: Betzig
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000196:embryology*; D002450:Cell Communication; D004331:Drosophila melanogaster / Q000196:embryology*; D004625:Embryo, Nonmammalian / Q000648:ultrastructure*; D053595:Embryonic Stem Cells / Q000648:ultrastructure; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008853:Microscopy / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D018874:Spheroids, Cellular / Q000648:ultrastructure