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Macromolecular 3D SEM reconstruction strategies: Signal to noise ratio and resolution
Three-dimensional scanning electron microscopy generates quantitative volumetric structural data from SEM images of macromolecules. This technique provides a quick and easy way to define the quaternary structure and handedness of protein complexes. Here, we apply a variety of preparation and imaging...
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| Published in: | Ultramicroscopy 2014-09, Vol.144, p.43-49 |
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| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c401t-4281af1c867e77a7eadce3261c7a9975fa15eef798296419ba885e107952c9373 |
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| cites | cdi_FETCH-LOGICAL-c401t-4281af1c867e77a7eadce3261c7a9975fa15eef798296419ba885e107952c9373 |
| container_end_page | 49 |
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| container_title | Ultramicroscopy |
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| creator | Woodward, J.D. Wepf, R.A. |
| description | Three-dimensional scanning electron microscopy generates quantitative volumetric structural data from SEM images of macromolecules. This technique provides a quick and easy way to define the quaternary structure and handedness of protein complexes. Here, we apply a variety of preparation and imaging methods to filamentous actin in order to explore the relationship between resolution, signal-to-noise ratio, structural preservation and dataset size. This information can be used to define successful imaging strategies for different applications.
•F-actin SEM datasets were collected using 8 different preparation/ imaging techniques.•Datasets were reconstructed by back projection and compared/analyzed•3DSEM actin reconstructions can be produced with |
| doi_str_mv | 10.1016/j.ultramic.2014.04.009 |
| format | article |
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•F-actin SEM datasets were collected using 8 different preparation/ imaging techniques.•Datasets were reconstructed by back projection and compared/analyzed•3DSEM actin reconstructions can be produced with <100 views of the asymmetric unit.•Negatively stained macromolecules can be reconstructed by 3DSEM to ~3nm resolution.</description><identifier>ISSN: 0304-3991</identifier><identifier>EISSN: 1879-2723</identifier><identifier>DOI: 10.1016/j.ultramic.2014.04.009</identifier><identifier>PMID: 24830764</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Actins - chemistry ; Actins - ultrastructure ; Algorithms ; Animals ; Filamentous actin ; Handedness ; Imaging ; Imaging, Three-Dimensional - methods ; Imaging, Three-Dimensional - statistics & numerical data ; Iterative helical real-space reconstruction ; Macromolecular ; Macromolecular Substances - chemistry ; Macromolecular Substances - ultrastructure ; Microscopy, Electron, Scanning - methods ; Microscopy, Electron, Scanning - statistics & numerical data ; Microscopy, Electron, Transmission - methods ; Microscopy, Electron, Transmission - statistics & numerical data ; Proteins ; Rabbits ; Reconstruction ; Scanning electron microscopy ; Signal to noise ratio ; Specimen Handling ; Strategy ; Three dimensional ; Three-dimensional reconstruction</subject><ispartof>Ultramicroscopy, 2014-09, Vol.144, p.43-49</ispartof><rights>2014 Elsevier B.V.</rights><rights>Copyright © 2014 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c401t-4281af1c867e77a7eadce3261c7a9975fa15eef798296419ba885e107952c9373</citedby><cites>FETCH-LOGICAL-c401t-4281af1c867e77a7eadce3261c7a9975fa15eef798296419ba885e107952c9373</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24830764$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Woodward, J.D.</creatorcontrib><creatorcontrib>Wepf, R.A.</creatorcontrib><title>Macromolecular 3D SEM reconstruction strategies: Signal to noise ratio and resolution</title><title>Ultramicroscopy</title><addtitle>Ultramicroscopy</addtitle><description>Three-dimensional scanning electron microscopy generates quantitative volumetric structural data from SEM images of macromolecules. This technique provides a quick and easy way to define the quaternary structure and handedness of protein complexes. Here, we apply a variety of preparation and imaging methods to filamentous actin in order to explore the relationship between resolution, signal-to-noise ratio, structural preservation and dataset size. This information can be used to define successful imaging strategies for different applications.
•F-actin SEM datasets were collected using 8 different preparation/ imaging techniques.•Datasets were reconstructed by back projection and compared/analyzed•3DSEM actin reconstructions can be produced with <100 views of the asymmetric unit.•Negatively stained macromolecules can be reconstructed by 3DSEM to ~3nm resolution.</description><subject>Actins - chemistry</subject><subject>Actins - ultrastructure</subject><subject>Algorithms</subject><subject>Animals</subject><subject>Filamentous actin</subject><subject>Handedness</subject><subject>Imaging</subject><subject>Imaging, Three-Dimensional - methods</subject><subject>Imaging, Three-Dimensional - statistics & numerical data</subject><subject>Iterative helical real-space reconstruction</subject><subject>Macromolecular</subject><subject>Macromolecular Substances - chemistry</subject><subject>Macromolecular Substances - ultrastructure</subject><subject>Microscopy, Electron, Scanning - methods</subject><subject>Microscopy, Electron, Scanning - statistics & numerical data</subject><subject>Microscopy, Electron, Transmission - methods</subject><subject>Microscopy, Electron, Transmission - statistics & numerical data</subject><subject>Proteins</subject><subject>Rabbits</subject><subject>Reconstruction</subject><subject>Scanning electron microscopy</subject><subject>Signal to noise ratio</subject><subject>Specimen Handling</subject><subject>Strategy</subject><subject>Three dimensional</subject><subject>Three-dimensional reconstruction</subject><issn>0304-3991</issn><issn>1879-2723</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><recordid>eNqFkE1r3DAQhkVoSTZp_0LQsRdvZ2RZHz21pElbSOghzVko8jhosa1Usgv599GySa-BgRmY552Bh7FzhC0Cqs-77Tou2U8xbAWg3EItsEdsg0bbRmjRvmMbaEE2rbV4wk5L2QEAgjTH7ERI04JWcsPubnzIaUojhXX0mbff-e3lDc8U0lyWvIYlppnXyS_0EKl84bfxYfYjXxKfUyzE6yYm7ue-hkoa133gA3s_-LHQx5d-xu6uLv9c_Gyuf__4dfHtugkScGmkMOgHDEZp0tpr8n2gVigM2luru8FjRzRoa4RVEu29N6YjBG07EWyr2zP26XD3Mae_K5XFTbEEGkc_U1qLQ60EdEqheBvtpFDSWAMVVQe0mikl0-Aec5x8fnIIbm_f7dyrfbe376AW2Bo8f_mx3k_U_4-96q7A1wNAVcq_SNmVEGkO1MdqfHF9im_9eAZ0HJkH</recordid><startdate>20140901</startdate><enddate>20140901</enddate><creator>Woodward, J.D.</creator><creator>Wepf, R.A.</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7SR</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>L7M</scope></search><sort><creationdate>20140901</creationdate><title>Macromolecular 3D SEM reconstruction strategies: Signal to noise ratio and resolution</title><author>Woodward, J.D. ; Wepf, R.A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c401t-4281af1c867e77a7eadce3261c7a9975fa15eef798296419ba885e107952c9373</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Actins - chemistry</topic><topic>Actins - ultrastructure</topic><topic>Algorithms</topic><topic>Animals</topic><topic>Filamentous actin</topic><topic>Handedness</topic><topic>Imaging</topic><topic>Imaging, Three-Dimensional - methods</topic><topic>Imaging, Three-Dimensional - statistics & numerical data</topic><topic>Iterative helical real-space reconstruction</topic><topic>Macromolecular</topic><topic>Macromolecular Substances - chemistry</topic><topic>Macromolecular Substances - ultrastructure</topic><topic>Microscopy, Electron, Scanning - methods</topic><topic>Microscopy, Electron, Scanning - statistics & numerical data</topic><topic>Microscopy, Electron, Transmission - methods</topic><topic>Microscopy, Electron, Transmission - statistics & numerical data</topic><topic>Proteins</topic><topic>Rabbits</topic><topic>Reconstruction</topic><topic>Scanning electron microscopy</topic><topic>Signal to noise ratio</topic><topic>Specimen Handling</topic><topic>Strategy</topic><topic>Three dimensional</topic><topic>Three-dimensional reconstruction</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Woodward, J.D.</creatorcontrib><creatorcontrib>Wepf, R.A.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Engineered Materials Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><jtitle>Ultramicroscopy</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Woodward, J.D.</au><au>Wepf, R.A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Macromolecular 3D SEM reconstruction strategies: Signal to noise ratio and resolution</atitle><jtitle>Ultramicroscopy</jtitle><addtitle>Ultramicroscopy</addtitle><date>2014-09-01</date><risdate>2014</risdate><volume>144</volume><spage>43</spage><epage>49</epage><pages>43-49</pages><issn>0304-3991</issn><eissn>1879-2723</eissn><abstract>Three-dimensional scanning electron microscopy generates quantitative volumetric structural data from SEM images of macromolecules. This technique provides a quick and easy way to define the quaternary structure and handedness of protein complexes. Here, we apply a variety of preparation and imaging methods to filamentous actin in order to explore the relationship between resolution, signal-to-noise ratio, structural preservation and dataset size. This information can be used to define successful imaging strategies for different applications.
•F-actin SEM datasets were collected using 8 different preparation/ imaging techniques.•Datasets were reconstructed by back projection and compared/analyzed•3DSEM actin reconstructions can be produced with <100 views of the asymmetric unit.•Negatively stained macromolecules can be reconstructed by 3DSEM to ~3nm resolution.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>24830764</pmid><doi>10.1016/j.ultramic.2014.04.009</doi><tpages>7</tpages></addata></record> |
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| identifier | ISSN: 0304-3991 |
| ispartof | Ultramicroscopy, 2014-09, Vol.144, p.43-49 |
| issn | 0304-3991 1879-2723 |
| language | eng |
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| source | ScienceDirect Freedom Collection |
| subjects | Actins - chemistry Actins - ultrastructure Algorithms Animals Filamentous actin Handedness Imaging Imaging, Three-Dimensional - methods Imaging, Three-Dimensional - statistics & numerical data Iterative helical real-space reconstruction Macromolecular Macromolecular Substances - chemistry Macromolecular Substances - ultrastructure Microscopy, Electron, Scanning - methods Microscopy, Electron, Scanning - statistics & numerical data Microscopy, Electron, Transmission - methods Microscopy, Electron, Transmission - statistics & numerical data Proteins Rabbits Reconstruction Scanning electron microscopy Signal to noise ratio Specimen Handling Strategy Three dimensional Three-dimensional reconstruction |
| title | Macromolecular 3D SEM reconstruction strategies: Signal to noise ratio and resolution |
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