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Science. 2014 Oct 24;346(6208):1257998. doi: 10.1126/science.1257998. Epub 2014 Oct 23.

Lattice light-sheet microscopy: imaging molecules to embryos at high spatiotemporal resolution.

Author information

1
Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.
2
Coleman Technologies, Incorporated, Newtown Square, PA 19073, USA.
3
National High Magnetic Field Laboratory and Department of Biological Science, Florida State University, Tallahassee, FL 32310, USA.
4
Department of Biological Sciences, University of the Sciences, Philadelphia, PA 19104, USA.
5
Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
6
Optical Image Analysis Unit, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan.
7
Department of Pharmacology, University of Minnesota, Minneapolis, MN 55455, USA.
8
Cell Biology and Metabolism Program, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA. Cambridge Institute for Medical Research, Addenbrooke's Hospital, Cambridge CB2 0XY, England, UK.
9
Cell Biology and Metabolism Program, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
10
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA.
11
Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA.
12
Max Planck Institute for Molecular Cell Biology and Genetics, 01307 Dresden, Germany. Max Planck Institute for the Physics of Complex Systems, 01307 Dresden, Germany.
13
Department of Molecular Biology and Genetics, Howard Hughes Medical Institute, Center for Cell Dynamics, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
14
Department of Biology, Duke University, Durham, NC 27708, USA.
15
Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA. betzige@janelia.hhmi.org.

Abstract

Although fluorescence microscopy provides a crucial window into the physiology of living specimens, many biological processes are too fragile, are too small, or occur too rapidly to see clearly with existing tools. We crafted ultrathin light sheets from two-dimensional optical lattices that allowed us to image three-dimensional (3D) dynamics for hundreds of volumes, often at subsecond intervals, at the diffraction limit and beyond. We applied this to systems spanning four orders of magnitude in space and time, including the diffusion of single transcription factor molecules in stem cell spheroids, the dynamic instability of mitotic microtubules, the immunological synapse, neutrophil motility in a 3D matrix, and embryogenesis in Caenorhabditis elegans and Drosophila melanogaster. The results provide a visceral reminder of the beauty and the complexity of living systems.

Comment in

PMID:
25342811
PMCID:
PMC4336192
DOI:
10.1126/science.1257998
[Indexed for MEDLINE]
Free PMC Article

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