Three-Dimensional scanning transmission electron microscopy of biological specimens

Handle URI:
http://hdl.handle.net/10754/561456
Title:
Three-Dimensional scanning transmission electron microscopy of biological specimens
Authors:
De Jonge, Niels; Sougrat, Rachid; Northan, Brian M.; Pennycook, Stephen J.
Abstract:
A three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM). The specimen was a metallic replica of the biological structure comprising Pt nanoparticles 2-3 nm in diameter, with a high stability under electron beam radiation. The 3D dataset was processed by an automated deconvolution procedure. The lateral resolution was 1.1 nm, set by pixel size. Particles differing by only 10 nm in vertical position were identified as separate objects with greater than 20% dip in contrast between them. We refer to this value as the axial resolution of the deconvolution or reconstruction, the ability to recognize two objects, which were unresolved in the original dataset. The resolution of the reconstruction is comparable to that achieved by tilt-series transmission electron microscopy. However, the focal-series method does not require mechanical tilting and is therefore much faster. 3D STEM images were also recorded of the Golgi ribbon in conventional thin sections containing 3T3 cells with a comparable axial resolution in the deconvolved dataset. © 2010 Microscopy Society of America.
KAUST Department:
Core Labs
Publisher:
Cambridge University Press (CUP)
Journal:
Microscopy and Microanalysis
Issue Date:
18-Jan-2010
DOI:
10.1017/S1431927609991280
PubMed ID:
20082729
PubMed Central ID:
PMC2917646
Type:
Article
ISSN:
14319276
Additional Links:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2917646
Appears in Collections:
Articles

Full metadata record

DC FieldValue Language
dc.contributor.authorDe Jonge, Nielsen
dc.contributor.authorSougrat, Rachiden
dc.contributor.authorNorthan, Brian M.en
dc.contributor.authorPennycook, Stephen J.en
dc.date.accessioned2015-08-02T09:11:50Zen
dc.date.available2015-08-02T09:11:50Zen
dc.date.issued2010-01-18en
dc.identifier.issn14319276en
dc.identifier.pmid20082729en
dc.identifier.doi10.1017/S1431927609991280en
dc.identifier.urihttp://hdl.handle.net/10754/561456en
dc.description.abstractA three-dimensional (3D) reconstruction of the cytoskeleton and a clathrin-coated pit in mammalian cells has been achieved from a focal-series of images recorded in an aberration-corrected scanning transmission electron microscope (STEM). The specimen was a metallic replica of the biological structure comprising Pt nanoparticles 2-3 nm in diameter, with a high stability under electron beam radiation. The 3D dataset was processed by an automated deconvolution procedure. The lateral resolution was 1.1 nm, set by pixel size. Particles differing by only 10 nm in vertical position were identified as separate objects with greater than 20% dip in contrast between them. We refer to this value as the axial resolution of the deconvolution or reconstruction, the ability to recognize two objects, which were unresolved in the original dataset. The resolution of the reconstruction is comparable to that achieved by tilt-series transmission electron microscopy. However, the focal-series method does not require mechanical tilting and is therefore much faster. 3D STEM images were also recorded of the Golgi ribbon in conventional thin sections containing 3T3 cells with a comparable axial resolution in the deconvolved dataset. © 2010 Microscopy Society of America.en
dc.publisherCambridge University Press (CUP)en
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC2917646en
dc.subjectAberration-corrected STEMen
dc.subjectBiological electron microscopyen
dc.subjectClathrin-coated piten
dc.subjectCytoskeletonen
dc.subjectDeconvolutionen
dc.subjectNanoparticlesen
dc.subjectThin sectionsen
dc.subjectThree-dimensional electron microscopyen
dc.titleThree-Dimensional scanning transmission electron microscopy of biological specimensen
dc.typeArticleen
dc.contributor.departmentCore Labsen
dc.identifier.journalMicroscopy and Microanalysisen
dc.identifier.pmcidPMC2917646en
dc.contributor.institutionVanderbilt University Medical Center, Department of Molecular Physiology and Biophysics, Light Hall 702, Nashville, TN 37232-0615, United Statesen
dc.contributor.institutionOak Ridge National Laboratory, Materials Science and Technology Division, 1 Bethel Valley Rd., Oak Ridge, TN 37831-6064, United Statesen
dc.contributor.institutionCell Biology and Metabolism Branch, NICHD, National Institute of Health, 18 Library Drive, Bethesda, MD 20892-5430, United Statesen
dc.contributor.institutionMedia Cybernetics Inc., 4340 East-West Hwy, Bethesda, MD 20814-4411, United Statesen
kaust.authorSougrat, Rachiden
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