DOI: 10.1126/science.1156947. Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy

Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy

10.1126/science.1156947

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

Fluorescence light microscopy allows multicolor visualization of cellular components with high specificity, but its utility has until recently been constrained by the intrinsic limit of spatial resolution. We applied three-dimensional structured illumination microscopy (3D-SIM) to circumvent this limit and to study the mammalian nucleus. By simultaneously imaging chromatin, nuclear lamina, and the nuclear pore complex (NPC), we observed several features that escape detection by conventional microscopy. We could resolve single NPCs that colocalized with channels in the lamin network and peripheral heterochromatin. We could differentially localize distinct NPC components and detect double-layered invaginations of the nuclear envelope in prophase as previously seen only by electron microscopy. Multicolor 3D-SIM opens new and facile possibilities to analyze subcellular structures beyond the diffraction limit of the emitted light.

Bibliography

Schermelleh, L., Carlton, P. M., Haase, S., Shao, L., Winoto, L., Kner, P., Burke, B., Cardoso, M. C., Agard, D. A., Gustafsson, M. G. L., Leonhardt, H., & Sedat, J. W. (2008). Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy. Science, 320(5881), 1332â1336.

Authors 12

Lothar Schermelleh (first)
Peter M. Carlton (additional)
Sebastian Haase (additional)
Lin Shao (additional)
Lukman Winoto (additional)
Peter Kner (additional)
Brian Burke (additional)
M. Cristina Cardoso (additional)
David A. Agard (additional)
Mats G. L. Gustafsson (additional)
Heinrich Leonhardt (additional)
John W. Sedat (additional)

References 30 Referenced 996

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This work was supported by grants from the Bavaria California Technology Center the Center for NanoScience the Nanosystems Initiative Munich and the Deutsche Forschungsgemeinschaft to L. Schermelleh M.C.C. and H.L.; by NIH grant GM-2501–25 to J.W.S.; by the David and Lucile Packard Foundation; and by NSF through the Center for Biophotonics an NSF Science and Technology Center managed by the University of California Davis under cooperative agreement no. PHY 0120999. P.M.C. is partially supported by the Keck Laboratory for Advanced Microscopy at the University of California San Francisco. We thank A. Čopïč K. Weis and F. Spada for comments on the manuscript and helpful discussions. P.M.C. L. Shao L.W. and P.K. have performed limited paid consulting for Applied Precision which is planning a commercial microscope system using three-dimensional structured illumination. The University of California holds patents for structured illumination microscopy.

Dates

Type	When
Created	17 years, 2 months ago (June 6, 2008, 2:16 p.m.)
Deposited	1 year, 7 months ago (Jan. 10, 2024, 3:41 a.m.)
Indexed	19 minutes ago (Aug. 26, 2025, 8:31 p.m.)
Issued	17 years, 2 months ago (June 6, 2008)
Published	17 years, 2 months ago (June 6, 2008)
Published Print	17 years, 2 months ago (June 6, 2008)

Funders 0

None

BibTeX

@article{Schermelleh_2008, title={Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy}, volume={320}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1156947}, DOI={10.1126/science.1156947}, number={5881}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Schermelleh, Lothar and Carlton, Peter M. and Haase, Sebastian and Shao, Lin and Winoto, Lukman and Kner, Peter and Burke, Brian and Cardoso, M. Cristina and Agard, David A. and Gustafsson, Mats G. L. and Leonhardt, Heinrich and Sedat, John W.}, year={2008}, month=jun, pages={1332–1336} }

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      "unstructured": "This work was supported by grants from the Bavaria California Technology Center the Center for NanoScience the Nanosystems Initiative Munich and the Deutsche Forschungsgemeinschaft to L. Schermelleh M.C.C. and H.L.; by NIH grant GM-2501\u201325 to J.W.S.; by the David and Lucile Packard Foundation; and by NSF through the Center for Biophotonics an NSF Science and Technology Center managed by the University of California Davis under cooperative agreement no. PHY 0120999. P.M.C. is partially supported by the Keck Laboratory for Advanced Microscopy at the University of California San Francisco. We thank A. \u010cop\u00ef\u010d K. Weis and F. Spada for comments on the manuscript and helpful discussions. P.M.C. L. Shao L.W. and P.K. have performed limited paid consulting for Applied Precision which is planning a commercial microscope system using three-dimensional structured illumination. The University of California holds patents for structured illumination microscopy."
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