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

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Molecular Biology of the Cell. 2016 Aug 17;27(22):3418-35. doi: 10.1091/mbc.E16-03-0164
Membrane dynamics of dividing cells imaged by lattice light-sheet microscopy. Betzig Lab
Aguet F, Upadhyayula S, Gaudin R, Chou Y, Cocucci E, He K, Chen B, Mosaliganti K, Pasham M, Skillern W, Legant WR, Liu T, Findlay G, Marino E, Danuser G, Megason S, Betzig E, Kirchhausen T
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Abstract

Membrane remodeling is an essential part for transfer of components to and from the cell surface and membrane-bound organelles, and for changes in cell shape, particularly critical during cell division. Earlier analyses, based on classical optical live-cell imaging, 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 non-invasive illumination of the newly developed lattice light sheet fluorescence microscope, we reexamined their assembly dynamics over the entire cell surface and showed 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 remained 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 in multicellular assemblies.

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Janelia Authors

Wesley Legant
Eric Betzig
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