Detection and imaging the expression of the trans-membrane protein CD44 in RT112 cells by use of enzyme-labeled antibodies and SECM
The deposition of human RT112 cells in a patterned fashion onto glass substrates and subsequent imaging of the expression of the trans-membrane protein CD44 have been studied using scanning electrochemical microscopy (SECM). Patterns of RT112 cells derived from a transitional cell carcinoma of the b...
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| Veröffentlicht in: | Biosensors & bioelectronics 2013-03, Vol.41, p.282-288 |
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| creator | Roberts, William S. Davis, Frank Holmes, Joanne L. Collyer, Stuart D. Larcombe, Lee D. Morgan, Sarah L. Higson, Séamus P.J. |
| description | The deposition of human RT112 cells in a patterned fashion onto glass substrates and subsequent imaging of the expression of the trans-membrane protein CD44 have been studied using scanning electrochemical microscopy (SECM). Patterns of RT112 cells derived from a transitional cell carcinoma of the bladder could be deposited on amino-modified glass substrates by cytospinning. These were then treated with horseradish peroxidase (HRP) labeled secondary antibodies to the trans-membrane protein CD44. Expression of CD44 protein by the cells directly leads to immobilisation of the labeled antibodies. The presence of the enzyme substrate (hydrogen peroxide) along with a hydroquinone mediator then allowed an enzymatic reaction to proceed, generating benzoquinone. Reduction of benzoquinone gave rise to positive feedback between the substrate and the SECM microelectrode tip. Control samples such as blank slides or slides not treated with HRP-labeled antibody showed negative feedback effects. Patterns of RT112 cells could be assembled and their expression of the target protein imaged whereas control samples showed minimal activity.
► RT112 cells in patterns on glass expressed the trans-membrane protein CD44. ► Expression of CD44 by the cells lead to immobilisation of enzyme-labeled antibodies. ► Enzymatic reactions allowed imaging of cells by scanning electrochemical microscopy. ► Patterns of RT112 cells assembled and their expression of the target protein imaged. ► Control samples showed minimal activity |
| doi_str_mv | 10.1016/j.bios.2012.08.038 |
| format | Article |
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► RT112 cells in patterns on glass expressed the trans-membrane protein CD44. ► Expression of CD44 by the cells lead to immobilisation of enzyme-labeled antibodies. ► Enzymatic reactions allowed imaging of cells by scanning electrochemical microscopy. ► Patterns of RT112 cells assembled and their expression of the target protein imaged. ► Control samples showed minimal activity</description><identifier>ISSN: 0956-5663</identifier><identifier>EISSN: 1873-4235</identifier><identifier>DOI: 10.1016/j.bios.2012.08.038</identifier><identifier>PMID: 23017674</identifier><language>eng</language><publisher>Kidlington: Elsevier B.V</publisher><subject>Biological and medical sciences ; Biotechnology ; CD44 ; Cell imaging ; Cell Line, Tumor ; Conductometry - methods ; Fundamental and applied biological sciences. Psychology ; Horseradish Peroxidase - chemistry ; Humans ; Hyaluronan Receptors - metabolism ; Microscopy, Electron, Scanning - methods ; Protein expression ; Scanning electrochemical microscopy ; Staining and Labeling - methods ; Urinary Bladder Neoplasms - metabolism ; Urinary Bladder Neoplasms - pathology</subject><ispartof>Biosensors & bioelectronics, 2013-03, Vol.41, p.282-288</ispartof><rights>2012 Elsevier B.V.</rights><rights>2014 INIST-CNRS</rights><rights>Copyright © 2012 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c423t-d4f86e81ec5c10bee57609365591ae111b5324b19656ed5f317570b54557cad93</citedby><cites>FETCH-LOGICAL-c423t-d4f86e81ec5c10bee57609365591ae111b5324b19656ed5f317570b54557cad93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bios.2012.08.038$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3541,27915,27916,45986</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=26924555$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23017674$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Roberts, William S.</creatorcontrib><creatorcontrib>Davis, Frank</creatorcontrib><creatorcontrib>Holmes, Joanne L.</creatorcontrib><creatorcontrib>Collyer, Stuart D.</creatorcontrib><creatorcontrib>Larcombe, Lee D.</creatorcontrib><creatorcontrib>Morgan, Sarah L.</creatorcontrib><creatorcontrib>Higson, Séamus P.J.</creatorcontrib><title>Detection and imaging the expression of the trans-membrane protein CD44 in RT112 cells by use of enzyme-labeled antibodies and SECM</title><title>Biosensors & bioelectronics</title><addtitle>Biosens Bioelectron</addtitle><description>The deposition of human RT112 cells in a patterned fashion onto glass substrates and subsequent imaging of the expression of the trans-membrane protein CD44 have been studied using scanning electrochemical microscopy (SECM). Patterns of RT112 cells derived from a transitional cell carcinoma of the bladder could be deposited on amino-modified glass substrates by cytospinning. These were then treated with horseradish peroxidase (HRP) labeled secondary antibodies to the trans-membrane protein CD44. Expression of CD44 protein by the cells directly leads to immobilisation of the labeled antibodies. The presence of the enzyme substrate (hydrogen peroxide) along with a hydroquinone mediator then allowed an enzymatic reaction to proceed, generating benzoquinone. Reduction of benzoquinone gave rise to positive feedback between the substrate and the SECM microelectrode tip. Control samples such as blank slides or slides not treated with HRP-labeled antibody showed negative feedback effects. Patterns of RT112 cells could be assembled and their expression of the target protein imaged whereas control samples showed minimal activity.
► RT112 cells in patterns on glass expressed the trans-membrane protein CD44. ► Expression of CD44 by the cells lead to immobilisation of enzyme-labeled antibodies. ► Enzymatic reactions allowed imaging of cells by scanning electrochemical microscopy. ► Patterns of RT112 cells assembled and their expression of the target protein imaged. ► Control samples showed minimal activity</description><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>CD44</subject><subject>Cell imaging</subject><subject>Cell Line, Tumor</subject><subject>Conductometry - methods</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Horseradish Peroxidase - chemistry</subject><subject>Humans</subject><subject>Hyaluronan Receptors - metabolism</subject><subject>Microscopy, Electron, Scanning - methods</subject><subject>Protein expression</subject><subject>Scanning electrochemical microscopy</subject><subject>Staining and Labeling - methods</subject><subject>Urinary Bladder Neoplasms - metabolism</subject><subject>Urinary Bladder Neoplasms - pathology</subject><issn>0956-5663</issn><issn>1873-4235</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU9v1DAQxS0EotvCF-CAfEHikuCxYzuRuKBtC0itKpVytmJ7UrzKn8XOIpYrX7xOd4Ebp7Hs92be_EzIK2AlMFDvNqUNUyo5A16yumSifkJWUGtRVFzIp2TFGqkKqZQ4IacpbRhjGhr2nJxwwUArXa3I73Oc0c1hGmk7ehqG9j6M93T-hhR_biOmtDxN3ePNHNsxFQMONh-QbuM0Yxjp-ryqaK63dwCcOuz7RO2e7hIuRhx_7Qcs-tZijz5PmYOdfMD0OPDLxfr6BXnWtX3Cl8d6Rr5eXtytPxVXNx8_rz9cFS7vMxe-6mqFNaCTDphFlFqxRigpG2gRAKwUvLLQKKnQy06AlppZWUmpXesbcUbeHvrm4N93mGYzhLTEzctMu2SAV6xuGq1VlvKD1MUppYid2cbMJu4NMLPANxuzwDcLfMNqk-Fn0-tj_50d0P-1_KGdBW-Ogja5tu8yRRfSP51qeA4rs-79QYeZxo-A0SQXcHToQ8yfZfwU_pfjAfiooXY</recordid><startdate>20130315</startdate><enddate>20130315</enddate><creator>Roberts, William S.</creator><creator>Davis, Frank</creator><creator>Holmes, Joanne L.</creator><creator>Collyer, Stuart D.</creator><creator>Larcombe, Lee D.</creator><creator>Morgan, Sarah L.</creator><creator>Higson, Séamus P.J.</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><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></search><sort><creationdate>20130315</creationdate><title>Detection and imaging the expression of the trans-membrane protein CD44 in RT112 cells by use of enzyme-labeled antibodies and SECM</title><author>Roberts, William S. ; Davis, Frank ; Holmes, Joanne L. ; Collyer, Stuart D. ; Larcombe, Lee D. ; Morgan, Sarah L. ; Higson, Séamus P.J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c423t-d4f86e81ec5c10bee57609365591ae111b5324b19656ed5f317570b54557cad93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>CD44</topic><topic>Cell imaging</topic><topic>Cell Line, Tumor</topic><topic>Conductometry - methods</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Horseradish Peroxidase - chemistry</topic><topic>Humans</topic><topic>Hyaluronan Receptors - metabolism</topic><topic>Microscopy, Electron, Scanning - methods</topic><topic>Protein expression</topic><topic>Scanning electrochemical microscopy</topic><topic>Staining and Labeling - methods</topic><topic>Urinary Bladder Neoplasms - metabolism</topic><topic>Urinary Bladder Neoplasms - pathology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Roberts, William S.</creatorcontrib><creatorcontrib>Davis, Frank</creatorcontrib><creatorcontrib>Holmes, Joanne L.</creatorcontrib><creatorcontrib>Collyer, Stuart D.</creatorcontrib><creatorcontrib>Larcombe, Lee D.</creatorcontrib><creatorcontrib>Morgan, Sarah L.</creatorcontrib><creatorcontrib>Higson, Séamus P.J.</creatorcontrib><collection>Pascal-Francis</collection><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><jtitle>Biosensors & bioelectronics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Roberts, William S.</au><au>Davis, Frank</au><au>Holmes, Joanne L.</au><au>Collyer, Stuart D.</au><au>Larcombe, Lee D.</au><au>Morgan, Sarah L.</au><au>Higson, Séamus P.J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection and imaging the expression of the trans-membrane protein CD44 in RT112 cells by use of enzyme-labeled antibodies and SECM</atitle><jtitle>Biosensors & bioelectronics</jtitle><addtitle>Biosens Bioelectron</addtitle><date>2013-03-15</date><risdate>2013</risdate><volume>41</volume><spage>282</spage><epage>288</epage><pages>282-288</pages><issn>0956-5663</issn><eissn>1873-4235</eissn><abstract>The deposition of human RT112 cells in a patterned fashion onto glass substrates and subsequent imaging of the expression of the trans-membrane protein CD44 have been studied using scanning electrochemical microscopy (SECM). Patterns of RT112 cells derived from a transitional cell carcinoma of the bladder could be deposited on amino-modified glass substrates by cytospinning. These were then treated with horseradish peroxidase (HRP) labeled secondary antibodies to the trans-membrane protein CD44. Expression of CD44 protein by the cells directly leads to immobilisation of the labeled antibodies. The presence of the enzyme substrate (hydrogen peroxide) along with a hydroquinone mediator then allowed an enzymatic reaction to proceed, generating benzoquinone. Reduction of benzoquinone gave rise to positive feedback between the substrate and the SECM microelectrode tip. Control samples such as blank slides or slides not treated with HRP-labeled antibody showed negative feedback effects. Patterns of RT112 cells could be assembled and their expression of the target protein imaged whereas control samples showed minimal activity.
► RT112 cells in patterns on glass expressed the trans-membrane protein CD44. ► Expression of CD44 by the cells lead to immobilisation of enzyme-labeled antibodies. ► Enzymatic reactions allowed imaging of cells by scanning electrochemical microscopy. ► Patterns of RT112 cells assembled and their expression of the target protein imaged. ► Control samples showed minimal activity</abstract><cop>Kidlington</cop><pub>Elsevier B.V</pub><pmid>23017674</pmid><doi>10.1016/j.bios.2012.08.038</doi><tpages>7</tpages></addata></record> |
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| subjects | Biological and medical sciences Biotechnology CD44 Cell imaging Cell Line, Tumor Conductometry - methods Fundamental and applied biological sciences. Psychology Horseradish Peroxidase - chemistry Humans Hyaluronan Receptors - metabolism Microscopy, Electron, Scanning - methods Protein expression Scanning electrochemical microscopy Staining and Labeling - methods Urinary Bladder Neoplasms - metabolism Urinary Bladder Neoplasms - pathology |
| title | Detection and imaging the expression of the trans-membrane protein CD44 in RT112 cells by use of enzyme-labeled antibodies and SECM |
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