| Noninvasive imaging beyond the diffraction limit of 3D dynamics in thickly fluorescent specimens. | |
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MedLine Citation:
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PMID: 23217717 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Optical imaging of the dynamics of living specimens involves tradeoffs between spatial resolution, temporal resolution, and phototoxicity, made more difficult in three dimensions. Here, however, we report that rapid three-dimensional (3D) dynamics can be studied beyond the diffraction limit in thick or densely fluorescent living specimens over many time points by combining ultrathin planar illumination produced by scanned Bessel beams with super-resolution structured illumination microscopy. We demonstrate in vivo karyotyping of chromosomes during mitosis and identify different dynamics for the actin cytoskeleton at the dorsal and ventral surfaces of fibroblasts. Compared to spinning disk confocal microscopy, we demonstrate substantially reduced photodamage when imaging rapid morphological changes in D. discoideum cells, as well as improved contrast and resolution at depth within developing C. elegans embryos. Bessel beam structured plane illumination thus promises new insights into complex biological phenomena that require 4D subcellular spatiotemporal detail in either a single or multicellular context. |
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Authors:
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Liang Gao; Lin Shao; Christopher D Higgins; John S Poulton; Mark Peifer; Michael W Davidson; Xufeng Wu; Bob Goldstein; Eric Betzig |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Cell Volume: 151 ISSN: 1097-4172 ISO Abbreviation: Cell Publication Date: 2012 Dec |
Date Detail:
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Created Date: 2012-12-10 Completed Date: 2013-02-06 Revised Date: 2013-06-11 |
Medline Journal Info:
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Nlm Unique ID: 0413066 Medline TA: Cell Country: United States |
Other Details:
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Languages: eng Pagination: 1370-85 Citation Subset: IM |
Copyright Information:
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Copyright © 2012 Elsevier Inc. All rights reserved. |
Affiliation:
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Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Brain / cytology, ultrastructure Caenorhabditis elegans / cytology, growth & development Cell Line Cell Line, Tumor Dermatitis, Phototoxic Dictyostelium / ultrastructure Drosophila melanogaster / cytology Fibroblasts / ultrastructure Humans Imaging, Three-Dimensional / methods* Karyotyping / methods Larva / cytology, ultrastructure Microscopy, Fluorescence / methods* Mitosis Optical Imaging / methods* |
| Grant Support | |
ID/Acronym/Agency:
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R01-GM083071/GM/NIGMS NIH HHS; R01-GM67236/GM/NIGMS NIH HHS; //Howard Hughes Medical Institute; //Howard Hughes Medical Institute |
| Comments/Corrections | |
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