title: Membrane dynamics of dividing cells imaged by lattice light-sheet microscopy.
abstract: Membrane remodeling is an essential part of transferring components to and from the cell surface and membrane-bound organelles and for changes in cell shape, which are particularly critical during cell division. Earlier analyses, based on classical optical live-cell imaging and mostly restricted by technical necessity to the attached bottom surface, showed persistent formation of endocytic clathrin pits and vesicles during mitosis. Taking advantage of the resolution, speed, and noninvasive illumination of the newly developed lattice light-sheet fluorescence microscope, we reexamined their assembly dynamics over the entire cell surface and found that clathrin pits form at a lower rate during late mitosis. Full-cell imaging measurements of cell surface area and volume throughout the cell cycle of single cells in culture and in zebrafish embryos showed that the total surface increased rapidly during the transition from telophase to cytokinesis, whereas cell volume increased slightly in metaphase and was relatively constant during cytokinesis. These applications demonstrate the advantage of lattice light-sheet microscopy and enable a new standard for imaging membrane dynamics in single cells and multicellular assemblies.
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115.
authors.forename: François
authors.initials: F
authors.lastname: Aguet
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
authors.forename: Srigokul
authors.initials: S
authors.lastname: Upadhyayula
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Gaudin
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
authors.forename: Yi-Ying
authors.initials: YY
authors.lastname: Chou
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
authors.forename: Emanuele
authors.initials: E
authors.lastname: Cocucci
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
authors.forename: Kangmin
authors.initials: K
authors.lastname: He
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147.
authors.forename: Bi-Chang
authors.initials: BC
authors.lastname: Chen
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115.
authors.forename: Kishore
authors.initials: K
authors.lastname: Mosaliganti
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
authors.forename: Mithun
authors.initials: M
authors.lastname: Pasham
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
authors.forename: Wesley
authors.initials: W
authors.lastname: Skillern
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147.
authors.forename: Wesley R
authors.initials: WR
authors.lastname: Legant
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147.
authors.forename: Tsung-Li
authors.initials: TL
authors.lastname: Liu
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
authors.forename: Greg
authors.initials: G
authors.lastname: Findlay
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
authors.forename: Eric
authors.initials: E
authors.lastname: Marino
authors.affiliation: Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, TX 75390.
authors.forename: Gaudenz
authors.initials: G
authors.lastname: Danuser
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115.
authors.forename: Sean
authors.initials: S
authors.lastname: Megason
authors.affiliation: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147.
authors.forename: Eric
authors.initials: E
authors.lastname: Betzig
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115 kirchhausen@crystal.harvard.edu.|Departments of Cell Biology and Pediatrics, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
authors.forename: Tom
authors.initials: T
authors.lastname: Kirchhausen
mesh_terms: D000818:Animals; D002453:Cell Cycle; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002966:Clathrin / Q000378:metabolism; D048749:Cytokinesis / Q000502:physiology; D011992:Endosomes / Q000378:metabolism; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008677:Metaphase; D008853:Microscopy / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D008938:Mitosis / Q000502:physiology; D015027:Zebrafish / Q000196:embryology
chemical_list: D002966:Clathrin