Quantification of protein based on single-molecule counting by total internal reflection fluorescence microscopy with adsorption equilibrium
We developed a sensitive single-molecule imaging method for quantification of protein by total internal reflection fluorescence microscopy with adsorption equilibrium. In this method, the adsorption equilibrium of protein was achieved between solution and glass substrate. Then, fluorescence images o...
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| Veröffentlicht in: | Analytica chimica acta 2007-05, Vol.590 (1), p.104-109 |
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| container_title | Analytica chimica acta |
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| creator | Wang, Lei Xu, Guang Shi, Zhikun Jiang, Wei Jin, Wenrui |
| description | We developed a sensitive single-molecule imaging method for quantification of protein by total internal reflection fluorescence microscopy with adsorption equilibrium. In this method, the adsorption equilibrium of protein was achieved between solution and glass substrate. Then, fluorescence images of protein molecules in a evanescent wave field were taken by a highly sensitive electron multiplying charge coupled device. Finally, the number of fluorescent spots corresponding to the protein molecules in the images was counted. Alexa Fluor 488-labeled goat anti-rat IgG(H
+
L) was chosen as the model protein. The spot number showed an excellent linear relationship with protein concentration. The concentration linear range was 5.4
×
10
−11 to 8.1
×
10
−10
mol
L
−1. |
| doi_str_mv | 10.1016/j.aca.2007.03.018 |
| format | Article |
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+
L) was chosen as the model protein. The spot number showed an excellent linear relationship with protein concentration. The concentration linear range was 5.4
×
10
−11 to 8.1
×
10
−10
mol
L
−1.</description><identifier>ISSN: 0003-2670</identifier><identifier>EISSN: 1873-4324</identifier><identifier>DOI: 10.1016/j.aca.2007.03.018</identifier><identifier>PMID: 17416229</identifier><identifier>CODEN: ACACAM</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Adsorption ; Analytical chemistry ; Chemistry ; Exact sciences and technology ; Immunoglobulin Heavy Chains - analysis ; Microscopy, Fluorescence - methods ; Protein ; Proteins - analysis ; Single-molecule detection ; Spectrometric and optical methods ; Total internal reflection fluorescence microscopy</subject><ispartof>Analytica chimica acta, 2007-05, Vol.590 (1), p.104-109</ispartof><rights>2007 Elsevier B.V.</rights><rights>2007 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c412t-ed61ca0ae7211f9f2eef61db391f612139571ef4fe9819d7d0abfa03ba564f8f3</citedby><cites>FETCH-LOGICAL-c412t-ed61ca0ae7211f9f2eef61db391f612139571ef4fe9819d7d0abfa03ba564f8f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0003267007005016$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18673741$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17416229$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Lei</creatorcontrib><creatorcontrib>Xu, Guang</creatorcontrib><creatorcontrib>Shi, Zhikun</creatorcontrib><creatorcontrib>Jiang, Wei</creatorcontrib><creatorcontrib>Jin, Wenrui</creatorcontrib><title>Quantification of protein based on single-molecule counting by total internal reflection fluorescence microscopy with adsorption equilibrium</title><title>Analytica chimica acta</title><addtitle>Anal Chim Acta</addtitle><description>We developed a sensitive single-molecule imaging method for quantification of protein by total internal reflection fluorescence microscopy with adsorption equilibrium. In this method, the adsorption equilibrium of protein was achieved between solution and glass substrate. Then, fluorescence images of protein molecules in a evanescent wave field were taken by a highly sensitive electron multiplying charge coupled device. Finally, the number of fluorescent spots corresponding to the protein molecules in the images was counted. Alexa Fluor 488-labeled goat anti-rat IgG(H
+
L) was chosen as the model protein. The spot number showed an excellent linear relationship with protein concentration. The concentration linear range was 5.4
×
10
−11 to 8.1
×
10
−10
mol
L
−1.</description><subject>Adsorption</subject><subject>Analytical chemistry</subject><subject>Chemistry</subject><subject>Exact sciences and technology</subject><subject>Immunoglobulin Heavy Chains - analysis</subject><subject>Microscopy, Fluorescence - methods</subject><subject>Protein</subject><subject>Proteins - analysis</subject><subject>Single-molecule detection</subject><subject>Spectrometric and optical methods</subject><subject>Total internal reflection fluorescence microscopy</subject><issn>0003-2670</issn><issn>1873-4324</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcGKFDEQhoMo7uzqA3iRXPTWbSrp6XTwJIu6woIIeg7pdGXNkE5mk25l3sGHNrMzsDc9VVJ8f1H1_4S8AtYCg_7drjXWtJwx2TLRMhiekA0MUjSd4N1TsmGMiYb3kl2Qy1J29cuBdc_JBcgOes7Vhvz5tpq4eOetWXyKNDm6z2lBH-loCk609oqPdwGbOQW0a0Bq01ol8Y6OB7qkxQTq44I51kdGV6GHSS6sKWOxGC3S2ducik37A_3tl5_UTCXl_QOH96sPfsx-nV-QZ86Egi_P9Yr8-PTx-_VNc_v185frD7eN7YAvDU49WMMMSg7glOOIrodpFApq5SDUVgK6zqEaQE1yYmZ0honRbPvODU5ckbenufXU-xXLomdfFw3BRExr0ZKJrVSs_y_IlQKm-LaCcAKPZ5bqgt5nP5t80MD0MSu90zUrfcxKM6FrVlXz-jx8HWecHhXncCrw5gyYYk1w2UTryyM39FJUtnLvTxxWz355zLpYf7R98rmGoafk_7HGXzk9tds</recordid><startdate>20070502</startdate><enddate>20070502</enddate><creator>Wang, Lei</creator><creator>Xu, Guang</creator><creator>Shi, Zhikun</creator><creator>Jiang, Wei</creator><creator>Jin, Wenrui</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>7U5</scope><scope>8FD</scope><scope>L7M</scope><scope>7X8</scope></search><sort><creationdate>20070502</creationdate><title>Quantification of protein based on single-molecule counting by total internal reflection fluorescence microscopy with adsorption equilibrium</title><author>Wang, Lei ; Xu, Guang ; Shi, Zhikun ; Jiang, Wei ; Jin, Wenrui</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c412t-ed61ca0ae7211f9f2eef61db391f612139571ef4fe9819d7d0abfa03ba564f8f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Adsorption</topic><topic>Analytical chemistry</topic><topic>Chemistry</topic><topic>Exact sciences and technology</topic><topic>Immunoglobulin Heavy Chains - analysis</topic><topic>Microscopy, Fluorescence - methods</topic><topic>Protein</topic><topic>Proteins - analysis</topic><topic>Single-molecule detection</topic><topic>Spectrometric and optical methods</topic><topic>Total internal reflection fluorescence microscopy</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Lei</creatorcontrib><creatorcontrib>Xu, Guang</creatorcontrib><creatorcontrib>Shi, Zhikun</creatorcontrib><creatorcontrib>Jiang, Wei</creatorcontrib><creatorcontrib>Jin, Wenrui</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>Solid State and Superconductivity Abstracts</collection><collection>Technology Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>MEDLINE - Academic</collection><jtitle>Analytica chimica acta</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Lei</au><au>Xu, Guang</au><au>Shi, Zhikun</au><au>Jiang, Wei</au><au>Jin, Wenrui</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantification of protein based on single-molecule counting by total internal reflection fluorescence microscopy with adsorption equilibrium</atitle><jtitle>Analytica chimica acta</jtitle><addtitle>Anal Chim Acta</addtitle><date>2007-05-02</date><risdate>2007</risdate><volume>590</volume><issue>1</issue><spage>104</spage><epage>109</epage><pages>104-109</pages><issn>0003-2670</issn><eissn>1873-4324</eissn><coden>ACACAM</coden><abstract>We developed a sensitive single-molecule imaging method for quantification of protein by total internal reflection fluorescence microscopy with adsorption equilibrium. In this method, the adsorption equilibrium of protein was achieved between solution and glass substrate. Then, fluorescence images of protein molecules in a evanescent wave field were taken by a highly sensitive electron multiplying charge coupled device. Finally, the number of fluorescent spots corresponding to the protein molecules in the images was counted. Alexa Fluor 488-labeled goat anti-rat IgG(H
+
L) was chosen as the model protein. The spot number showed an excellent linear relationship with protein concentration. The concentration linear range was 5.4
×
10
−11 to 8.1
×
10
−10
mol
L
−1.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>17416229</pmid><doi>10.1016/j.aca.2007.03.018</doi><tpages>6</tpages></addata></record> |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Adsorption Analytical chemistry Chemistry Exact sciences and technology Immunoglobulin Heavy Chains - analysis Microscopy, Fluorescence - methods Protein Proteins - analysis Single-molecule detection Spectrometric and optical methods Total internal reflection fluorescence microscopy |
| title | Quantification of protein based on single-molecule counting by total internal reflection fluorescence microscopy with adsorption equilibrium |
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