Document Detail

Noninvasive imaging beyond the diffraction limit of 3D dynamics in thickly fluorescent specimens.
MedLine Citation:
PMID:  23217717     Owner:  NLM     Status:  MEDLINE    
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.
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:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cell     Volume:  151     ISSN:  1097-4172     ISO Abbreviation:  Cell     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-10     Completed Date:  2013-02-06     Revised Date:  2014-06-11    
Medline Journal Info:
Nlm Unique ID:  0413066     Medline TA:  Cell     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1370-85     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 Elsevier Inc. All rights reserved.
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MeSH Terms
Brain / cytology,  ultrastructure
Caenorhabditis elegans / cytology,  growth & development
Cell Line
Cell Line, Tumor
Dermatitis, Phototoxic
Dictyostelium / ultrastructure
Drosophila melanogaster / cytology
Fibroblasts / ultrastructure
Imaging, Three-Dimensional / methods*
Karyotyping / methods
Larva / cytology,  ultrastructure
Microscopy, Fluorescence / methods*
Optical Imaging / methods*
Grant Support
R01 GM067236/GM/NIGMS NIH HHS; R01 GM083071/GM/NIGMS NIH HHS; R01-GM083071/GM/NIGMS NIH HHS; R01-GM67236/GM/NIGMS NIH HHS; //Howard Hughes Medical Institute; //Howard Hughes Medical Institute

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