Single-cell lysis for visual analysis by electron microscopy.
Identifieur interne : 000398 ( Main/Exploration ); précédent : 000397; suivant : 000399Single-cell lysis for visual analysis by electron microscopy.
Auteurs : RBID : pubmed:23816812English descriptors
- KwdEn :
- MESH :
- metabolism : Fibroblasts.
- methods : Single-Cell Analysis.
- ultrastructure : Fibroblasts, Organelles.
- Animals, Cell Line, Cricetinae, Microscopy, Electron, Transmission.
Abstract
The stochastic nature of biological systems makes the study of individual cells a necessity in systems biology. Yet, handling and disruption of single cells and the analysis of the relatively low concentrations of their protein components still challenges available techniques. Transmission electron microscopy (TEM) allows for the analysis of proteins at the single-molecule level. Here, we present a system for single-cell lysis under light microscopy observation, followed by rapid uptake of the cell lysate. Eukaryotic cells were grown on conductively coated glass slides and observed by light microscopy. A custom-designed microcapillary electrode was used to target and lyse individual cells with electrical pulses. Nanoliter volumes were subsequently aspirated into the microcapillary and dispensed onto an electron microscopy grid for TEM inspection. We show, that the cell lysis and preparation method conserves protein structures well and is suitable for visual analysis by TEM.
DOI: 10.1016/j.jsb.2013.06.012
PubMed: 23816812
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Single-cell lysis for visual analysis by electron microscopy.</title><author><name sortKey="Kemmerling, Simon" uniqKey="Kemmerling S">Simon Kemmerling</name><affiliation wicri:level="1"><nlm:affiliation>Center for Cellular Imaging and Nano Analytics (C-CINA), Biozentrum, University of Basel, Basel, Switzerland.</nlm:affiliation><country xml:lang="fr">Suisse</country><wicri:regionArea>Center for Cellular Imaging and Nano Analytics (C-CINA), Biozentrum, University of Basel, Basel</wicri:regionArea></affiliation></author><author><name sortKey="Arnold, Stefan A" uniqKey="Arnold S">Stefan A Arnold</name></author><author><name sortKey="Bircher, Benjamin A" uniqKey="Bircher B">Benjamin A Bircher</name></author><author><name sortKey="Sauter, Nora" uniqKey="Sauter N">Nora Sauter</name></author><author><name sortKey="Escobedo, Carlos" uniqKey="Escobedo C">Carlos Escobedo</name></author><author><name sortKey="Dernick, Gregor" uniqKey="Dernick G">Gregor Dernick</name></author><author><name sortKey="Hierlemann, Andreas" uniqKey="Hierlemann A">Andreas Hierlemann</name></author><author><name sortKey="Stahlberg, Henning" uniqKey="Stahlberg H">Henning Stahlberg</name></author><author><name sortKey="Braun, Thomas" uniqKey="Braun T">Thomas Braun</name></author></titleStmt><publicationStmt><date when="2013">2013</date><idno type="RBID">pubmed:23816812</idno><idno type="pmid">23816812</idno><idno type="doi">10.1016/j.jsb.2013.06.012</idno><idno type="wicri:Area/Main/Corpus">000546</idno><idno type="wicri:Area/Main/Curation">000546</idno><idno type="wicri:Area/Main/Exploration">000398</idno></publicationStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Cell Line</term><term>Cricetinae</term><term>Fibroblasts (metabolism)</term><term>Fibroblasts (ultrastructure)</term><term>Microscopy, Electron, Transmission</term><term>Organelles (ultrastructure)</term><term>Single-Cell Analysis (methods)</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Fibroblasts</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Single-Cell Analysis</term></keywords><keywords scheme="MESH" qualifier="ultrastructure" xml:lang="en"><term>Fibroblasts</term><term>Organelles</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cell Line</term><term>Cricetinae</term><term>Microscopy, Electron, Transmission</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The stochastic nature of biological systems makes the study of individual cells a necessity in systems biology. Yet, handling and disruption of single cells and the analysis of the relatively low concentrations of their protein components still challenges available techniques. Transmission electron microscopy (TEM) allows for the analysis of proteins at the single-molecule level. Here, we present a system for single-cell lysis under light microscopy observation, followed by rapid uptake of the cell lysate. Eukaryotic cells were grown on conductively coated glass slides and observed by light microscopy. A custom-designed microcapillary electrode was used to target and lyse individual cells with electrical pulses. Nanoliter volumes were subsequently aspirated into the microcapillary and dispensed onto an electron microscopy grid for TEM inspection. We show, that the cell lysis and preparation method conserves protein structures well and is suitable for visual analysis by TEM.</div></front></TEI><pubmed><MedlineCitation Owner="NLM" Status="MEDLINE"><PMID Version="1">23816812</PMID><DateCreated><Year>2013</Year><Month>09</Month><Day>09</Day></DateCreated><DateCompleted><Year>2014</Year><Month>03</Month><Day>20</Day></DateCompleted><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1095-8657</ISSN><JournalIssue CitedMedium="Internet"><Volume>183</Volume><Issue>3</Issue><PubDate><Year>2013</Year><Month>Sep</Month></PubDate></JournalIssue><Title>Journal of structural biology</Title><ISOAbbreviation>J. Struct. Biol.</ISOAbbreviation></Journal><ArticleTitle>Single-cell lysis for visual analysis by electron microscopy.</ArticleTitle><Pagination><MedlinePgn>467-73</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.jsb.2013.06.012</ELocationID><ELocationID EIdType="pii" ValidYN="Y">S1047-8477(13)00167-6</ELocationID><Abstract><AbstractText>The stochastic nature of biological systems makes the study of individual cells a necessity in systems biology. Yet, handling and disruption of single cells and the analysis of the relatively low concentrations of their protein components still challenges available techniques. Transmission electron microscopy (TEM) allows for the analysis of proteins at the single-molecule level. Here, we present a system for single-cell lysis under light microscopy observation, followed by rapid uptake of the cell lysate. Eukaryotic cells were grown on conductively coated glass slides and observed by light microscopy. A custom-designed microcapillary electrode was used to target and lyse individual cells with electrical pulses. Nanoliter volumes were subsequently aspirated into the microcapillary and dispensed onto an electron microscopy grid for TEM inspection. We show, that the cell lysis and preparation method conserves protein structures well and is suitable for visual analysis by TEM.</AbstractText><CopyrightInformation>Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Kemmerling</LastName><ForeName>Simon</ForeName><Initials>S</Initials><Affiliation>Center for Cellular Imaging and Nano Analytics (C-CINA), Biozentrum, University of Basel, Basel, Switzerland.</Affiliation></Author><Author ValidYN="Y"><LastName>Arnold</LastName><ForeName>Stefan A</ForeName><Initials>SA</Initials></Author><Author ValidYN="Y"><LastName>Bircher</LastName><ForeName>Benjamin A</ForeName><Initials>BA</Initials></Author><Author ValidYN="Y"><LastName>Sauter</LastName><ForeName>Nora</ForeName><Initials>N</Initials></Author><Author ValidYN="Y"><LastName>Escobedo</LastName><ForeName>Carlos</ForeName><Initials>C</Initials></Author><Author ValidYN="Y"><LastName>Dernick</LastName><ForeName>Gregor</ForeName><Initials>G</Initials></Author><Author ValidYN="Y"><LastName>Hierlemann</LastName><ForeName>Andreas</ForeName><Initials>A</Initials></Author><Author 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