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dc.contributor.authorDahmke, Indra N
dc.contributor.authorVerch, Andreas
dc.contributor.authorHermannsdörfer, Justus
dc.contributor.authorPeckys, Diana B
dc.contributor.authorWeatherup, Robert S
dc.contributor.authorHofmann, Stephan
dc.contributor.authorde Jonge, Niels
dc.date.accessioned2018-05-04T10:42:12Z
dc.date.available2018-05-04T10:42:12Z
dc.date.issued2017-11-28
dc.identifier.issn1936-0851
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/275562
dc.description.abstractMembrane proteins govern many important functions in cells via dynamic oligomerization into active complexes. However, analytical methods to study their distribution and functional state in relation to the cellular structure are currently limited. Here, we introduce a technique for studying single-membrane proteins within their native context of the intact plasma membrane. SKBR3 breast cancer cells were grown on silicon microchips with thin silicon nitride windows. The cells were fixed, and the epidermal growth factor receptor ErbB2 was specifically labeled with quantum dot (QD) nanoparticles. For correlative fluorescence- and liquid-phase electron microscopy, we enclosed the liquid samples by chemical vapor deposited (CVD) graphene films. Depending on the local cell thickness, QD labels were imaged with a spatial resolution of 2 nm at a low electron dose. The distribution and stoichiometric assembly of ErbB2 receptors were determined at several different cellular locations, including tunneling nanotubes, where we found higher levels of homodimerization at the connecting sites. This experimental approach is applicable to a wide range of cell lines and membrane proteins and particularly suitable for studies involving both inter- and intracellular heterogeneity in protein distribution and expression.
dc.format.mediumPrint-Electronic
dc.languageeng
dc.publisherAmerican Chemical Society (ACS)
dc.subjectCell Line, Tumor
dc.subjectHumans
dc.subjectGraphite
dc.subjectSilicon Compounds
dc.subjectReceptor, erbB-2
dc.subjectMembrane Proteins
dc.subjectNeoplasm Proteins
dc.subjectMicroscopy, Electron
dc.subjectQuantum Dots
dc.subjectLab-On-A-Chip Devices
dc.subjectSingle Molecule Imaging
dc.titleGraphene Liquid Enclosure for Single-Molecule Analysis of Membrane Proteins in Whole Cells Using Electron Microscopy.
dc.typeArticle
prism.endingPage11117
prism.issueIdentifier11
prism.publicationDate2017
prism.publicationNameACS Nano
prism.startingPage11108
prism.volume11
dc.identifier.doi10.17863/CAM.22806
rioxxterms.versionofrecord10.1021/acsnano.7b05258
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2017-11
dc.contributor.orcidWeatherup, Robert S [0000-0002-3993-9045]
dc.contributor.orcidHofmann, Stephan [0000-0001-6375-1459]
dc.contributor.orcidde Jonge, Niels [0000-0002-3969-6821]
dc.identifier.eissn1936-086X
rioxxterms.typeJournal Article/Review
pubs.funder-project-idEngineering and Physical Sciences Research Council (EP/K016636/1)
pubs.funder-project-idEuropean Commission Horizon 2020 (H2020) Marie Sk?odowska-Curie actions (656870)
cam.issuedOnline2017-10-16
rioxxterms.freetoread.startdate2018-10-12


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