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Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination
Open accessAbstract
A key challenge when imaging living cells is how to noninvasively extract the most spatiotemporal information possible. Unlike popular wide-field and confocal methods, plane-illumination microscopy limits excitation to the information-rich vicinity of the focal plane, providing effective optical sectioning and high speed while minimizing out-of-focus background and premature photobleaching. Here we used scanned Bessel beams in conjunction with structured illumination and/or two-photon excitation to create thinner light sheets (<0.5 μm) better suited to three-dimensional (3D) subcellular imaging. As demonstrated by imaging the dynamics of mitochondria, filopodia, membrane ruffles, intracellular vesicles and mitotic chromosomes in live cells, the microscope currently offers 3D isotropic resolution down to ∼0.3 μm, speeds up to nearly 200 image planes per second and the ability to noninvasively acquire hundreds of 3D data volumes from single living cells encompassing tens of thousands of image frames.
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Cited by
Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution
Chen, B.-C., Legant, W. R., Wang, K., Shao, L., Milkie, D. E., Davidson, M. W., Janetopoulos, C., Wu, X. S., Hammer, J. A., Liu, Z., et al. (2014) · Science · doi:10.1126/science.1257998 · Cited by 1,919
Self-citationRichardson, D. S., & Lichtman, J. W. (2015) · Cell · doi:10.1016/j.cell.2015.06.067 · Cited by 1,095
Advanced CLARITY for rapid and high-resolution imaging of intact tissues
Tomer, R., Ye, L., Hsueh, B., & Deisseroth, K. (2014) · Nature Protocols · doi:10.1038/nprot.2014.123 · Cited by 842
CLARITY for mapping the nervous system
Chung, K., & Deisseroth, K. (2013) · Nature Methods · doi:10.1038/nmeth.2481 · Cited by 747
Single-Molecule Dynamics of Enhanceosome Assembly in Embryonic Stem Cells
Chen, J., Zhang, Z., Li, L., Chen, B.-C., Revyakin, A., Hajj, B., Legant, W., Dahan, M., Lionnet, T., Betzig, E., et al. (2014) · Cell · doi:10.1016/j.cell.2014.01.062 · Cited by 602
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Metadata
- Authors
- Planchon, T. A., Gao, L., Milkie, D. E., Davidson, M. W., Galbraith, J. A., Galbraith, C. G., & Betzig, E.
- Venue
- Nature Methods
- Published
- 2011-03-04
- Volume
- 8
- Issue
- 5
- Pages
- 417-423
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- OpenAlex
- W2168283408
- Cited by
- 1,112
- References
- 0
- FWCI
- 43.91
- Type
- journal article
- Language
- en
- Open access
- green
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- Planchon, T. A., Gao, L., Milkie, D. E., Davidson, M. W., Galbraith, J. A., Galbraith, C. G., & Betzig, E. (2011). Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination. Nature Methods. https://doi.org/10.1038/nmeth.1586
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