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Title:
Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy
Authors:
Schermelleh, Lothar; Carlton, Peter M.; Haase, Sebastian; Shao, Lin; Winoto, Lukman; Kner, Peter; Burke, Brian; Cardoso, M. Cristina; Agard, David A.; Gustafsson, Mats G. L.; Leonhardt, Heinrich; Sedat, John W.
Affiliation:
AA(Center for Integrated Protein Science, Department of Biology, Ludwig Maximilians University Munich, 82152 Planegg-Martinsried, Germany.), AB(Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA.), AC(Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA.; Max Delbrück Center for Molecular Medicine, 13125 Berlin, Germany.), AD(Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA.), AE(Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA.), AF(Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA.), AG(Department of Anatomy and Cell Biology, University of Florida, Gainesville, FL 32610, USA.), AH(Max Delbrück Center for Molecular Medicine, 13125 Berlin, Germany.), AI(Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA.), AJ(Department of Physiology and Program in Bioengineering, University of California, San Francisco, CA 94143, USA.), AK(Center for Integrated Protein Science, Department of Biology, Ludwig Maximilians University Munich, 82152 Planegg-Martinsried, Germany.), AL(Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA.)
Publication:
Science, Volume 320, Issue 5881, pp. 1332- (2008).
Publication Date:
06/2008
Category:
BIOCHEM
Origin:
SCIENCE
Abstract Copyright:
(c) 2008: Science
DOI:
10.1126/science.1156947
Bibliographic Code:
2008Sci...320.1332S

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.
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