Membrane dynamics of dividing cells imaged by lattice light-sheet microscopy

François Aguet, Srigokul Upadhyayula, Raphaël Gaudin, Yi Ying Chou, Emanuele Cocucci, Kangmin He, Bi Chang Chen, Kishore Mosaliganti, Mithun Pasham, Wesley Skillern, Wesley R. Legant, Tsung Li Liu, Greg Findlay, Eric Marino, Gaudenz Danuser, Sean Megason, Eric Betzig, Tom Kirchhausen

Research output: Contribution to journalArticle

36 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)3418-3435
Number of pages18
JournalMolecular Biology of the Cell
Volume27
Issue number22
DOIs
StatePublished - Nov 7 2016

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ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Aguet, F., Upadhyayula, S., Gaudin, R., Chou, Y. Y., Cocucci, E., He, K., Chen, B. C., Mosaliganti, K., Pasham, M., Skillern, W., Legant, W. R., Liu, T. L., Findlay, G., Marino, E., Danuser, G., Megason, S., Betzig, E., & Kirchhausen, T. (2016). Membrane dynamics of dividing cells imaged by lattice light-sheet microscopy. Molecular Biology of the Cell, 27(22), 3418-3435. https://doi.org/10.1091/mbc.E16-03-0164