The Effect of Electron Beam Irradiation in Environmental Scanning Transmission Electron Microscopy of Whole Cells in Liquid
Whole cells can be studied in their native liquid environment using electron microscopy, and unique information about the locations and stoichiometry of individual membrane proteins can be obtained from many cells thus taking cell heterogeneity into account. Of key importance for the further develop...
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| Veröffentlicht in: | Microscopy and microanalysis 2016-06, Vol.22 (3), p.656-665 |
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| creator | Hermannsdörfer, Justus Tinnemann, Verena Peckys, Diana B. de Jonge, Niels |
| description | Whole cells can be studied in their native liquid environment using electron microscopy, and unique information about the locations and stoichiometry of individual membrane proteins can be obtained from many cells thus taking cell heterogeneity into account. Of key importance for the further development of this microscopy technology is knowledge about the effect of electron beam radiation on the samples under investigation. We used environmental scanning electron microscopy (ESEM) with scanning transmission electron microscopy (STEM) detection to examine the effect of radiation for whole fixed COS7 fibroblasts in liquid. The main observation was the localization of nanoparticle labels attached to epidermal growth factor receptors (EGFRs). It was found that the relative distances between the labels remained mostly unchanged ( |
| doi_str_mv | 10.1017/S1431927616000763 |
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Of key importance for the further development of this microscopy technology is knowledge about the effect of electron beam radiation on the samples under investigation. We used environmental scanning electron microscopy (ESEM) with scanning transmission electron microscopy (STEM) detection to examine the effect of radiation for whole fixed COS7 fibroblasts in liquid. The main observation was the localization of nanoparticle labels attached to epidermal growth factor receptors (EGFRs). It was found that the relative distances between the labels remained mostly unchanged (<1.5%) for electron doses ranging from the undamaged native state at 10 e−/Å2 toward 103 e−/Å2. This dose range was sufficient to determine the EGFR locations with nanometer resolution and to distinguish between monomers and dimers. 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Of key importance for the further development of this microscopy technology is knowledge about the effect of electron beam radiation on the samples under investigation. We used environmental scanning electron microscopy (ESEM) with scanning transmission electron microscopy (STEM) detection to examine the effect of radiation for whole fixed COS7 fibroblasts in liquid. The main observation was the localization of nanoparticle labels attached to epidermal growth factor receptors (EGFRs). It was found that the relative distances between the labels remained mostly unchanged (<1.5%) for electron doses ranging from the undamaged native state at 10 e−/Å2 toward 103 e−/Å2. This dose range was sufficient to determine the EGFR locations with nanometer resolution and to distinguish between monomers and dimers. Various different forms of radiation damage became visible at higher doses, including severe dislocation, and the dissolution of labels.</description><subject>Animals</subject><subject>Biological Applications</subject><subject>Cells - radiation effects</subject><subject>Cells - ultrastructure</subject><subject>Cercopithecus aethiops</subject><subject>COS Cells</subject><subject>Electron microscopes</subject><subject>Environmental scanning</subject><subject>Epidermal growth factor</subject><subject>Fourier transforms</subject><subject>Heterogeneity</subject><subject>Irradiation</subject><subject>Microscopy, Electron, Scanning Transmission - instrumentation</subject><subject>Microscopy, Electron, Scanning Transmission - methods</subject><subject>Nanoparticles</subject><subject>Nanoparticles - chemistry</subject><subject>Proteins</subject><subject>Radiation</subject><subject>Scanning electron microscopy</subject><subject>Semiconductors</subject><subject>Silicon nitride</subject><subject>Tomography, X-Ray Computed</subject><subject>Transmission electron microscopy</subject><issn>1431-9276</issn><issn>1435-8115</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkd1r2zAQwMVoWT-2P2AvQ9CXvbjVh-2zH9eQtoGUPiRjj0axTomCLaeSPSj75ye3aRgbpSDQcffTTycdIV84u-SMw9WCp5KXAnKeM8Yglx_IaUxlScF5dvQc82Ssn5CzELaRkZH6SE4EcAkM4JT8Xm6QTo3BuqedodMmBr5z9BpVS2feK21Vb2PCOjp1v2ysteh61dBFrZyzbk2XXrnQ2hBG7CC4t7XvQt3tnkbvz03XIJ1g04TRNLePg9WfyLFRTcDP-_2c_LiZLid3yfzhdjb5Pk_qFESfAGBaqBIwg0xxWRgsQMNKZ6zQAovc6FqiyARjK1aWmAsDjClVG5RapDyX5-Tbi3fnu8cBQ1_FbuvYi3LYDaHiBSvyLE1L_j4KZSlYXDKiF_-g227wLj5kpKIQZMYixV-o8TeCR1PtvG2Vf6o4q8YhVv8NMZ75ujcPqxb14cTr1CIg91LVrrzVa_zr7je1fwCHtqXL</recordid><startdate>20160601</startdate><enddate>20160601</enddate><creator>Hermannsdörfer, Justus</creator><creator>Tinnemann, Verena</creator><creator>Peckys, Diana B.</creator><creator>de Jonge, Niels</creator><general>Cambridge University Press</general><general>Oxford University Press</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>3V.</scope><scope>7QO</scope><scope>7RV</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB0</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope></search><sort><creationdate>20160601</creationdate><title>The Effect of Electron Beam Irradiation in Environmental Scanning Transmission Electron Microscopy of Whole Cells in Liquid</title><author>Hermannsdörfer, Justus ; Tinnemann, Verena ; Peckys, Diana B. ; de Jonge, Niels</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c472t-77e48a97e575a138fe87d7bd508d2e86fdc3e25200b099e62f700aacfe3d24163</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Animals</topic><topic>Biological Applications</topic><topic>Cells - radiation effects</topic><topic>Cells - ultrastructure</topic><topic>Cercopithecus aethiops</topic><topic>COS Cells</topic><topic>Electron microscopes</topic><topic>Environmental scanning</topic><topic>Epidermal growth factor</topic><topic>Fourier transforms</topic><topic>Heterogeneity</topic><topic>Irradiation</topic><topic>Microscopy, Electron, Scanning Transmission - instrumentation</topic><topic>Microscopy, Electron, Scanning Transmission - methods</topic><topic>Nanoparticles</topic><topic>Nanoparticles - chemistry</topic><topic>Proteins</topic><topic>Radiation</topic><topic>Scanning electron microscopy</topic><topic>Semiconductors</topic><topic>Silicon nitride</topic><topic>Tomography, X-Ray Computed</topic><topic>Transmission electron microscopy</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hermannsdörfer, Justus</creatorcontrib><creatorcontrib>Tinnemann, Verena</creatorcontrib><creatorcontrib>Peckys, Diana B.</creatorcontrib><creatorcontrib>de Jonge, Niels</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Neurosciences Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>MEDLINE - 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Of key importance for the further development of this microscopy technology is knowledge about the effect of electron beam radiation on the samples under investigation. We used environmental scanning electron microscopy (ESEM) with scanning transmission electron microscopy (STEM) detection to examine the effect of radiation for whole fixed COS7 fibroblasts in liquid. The main observation was the localization of nanoparticle labels attached to epidermal growth factor receptors (EGFRs). It was found that the relative distances between the labels remained mostly unchanged (<1.5%) for electron doses ranging from the undamaged native state at 10 e−/Å2 toward 103 e−/Å2. This dose range was sufficient to determine the EGFR locations with nanometer resolution and to distinguish between monomers and dimers. 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| subjects | Animals Biological Applications Cells - radiation effects Cells - ultrastructure Cercopithecus aethiops COS Cells Electron microscopes Environmental scanning Epidermal growth factor Fourier transforms Heterogeneity Irradiation Microscopy, Electron, Scanning Transmission - instrumentation Microscopy, Electron, Scanning Transmission - methods Nanoparticles Nanoparticles - chemistry Proteins Radiation Scanning electron microscopy Semiconductors Silicon nitride Tomography, X-Ray Computed Transmission electron microscopy |
| title | The Effect of Electron Beam Irradiation in Environmental Scanning Transmission Electron Microscopy of Whole Cells in Liquid |
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