Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations
Extracellular vesicles (EVs), including exosomes, are circulating nanoscale particles heavily implicated in cell signaling and can be isolated in vast numbers from human biofluids. Study of their molecular profiling and materials properties is currently underway for purposes of describing a variety...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2017-05, Vol.89 (10), p.5357-5363 |
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| creator | Carney, Randy P Hazari, Sidhartha Colquhoun, Macalistair Tran, Di Hwang, Billanna Mulligan, Michael S Bryers, James D Girda, Eugenia Leiserowitz, Gary S Smith, Zachary J Lam, Kit S |
| description | Extracellular vesicles (EVs), including exosomes, are circulating nanoscale particles heavily implicated in cell signaling and can be isolated in vast numbers from human biofluids. Study of their molecular profiling and materials properties is currently underway for purposes of describing a variety of biological functions and diseases. However, the large, and as yet largely unquantified, variety of EV subpopulations differing in composition, size, and likely function necessitates characterization schemes capable of measuring single vesicles. Here we describe the first application of multispectral optical tweezers (MS-OTs) to single vesicles for molecular fingerprinting of EV subpopulations. This versatile imaging platform allows for sensitive measurement of Raman chemical composition (e.g., variation in protein, lipid, cholesterol, nucleic acids), coupled with discrimination by fluorescence markers. For exosomes isolated by ultracentrifugation, we use MS-OTs to interrogate the CD9-positive subpopulations via antibody fluorescence labeling and Raman spectra measurement. We report that the CD9-positive exosome subset exhibits reduced component concentration per vesicle and reduced chemical heterogeneity compared to the total purified EV population. We observed that specific vesicle subpopulations are present across exosomes isolated from cell culture supernatant of several clonal varieties of mesenchymal stromal cells and also from plasma and ascites isolated from human ovarian cancer patients. |
| doi_str_mv | 10.1021/acs.analchem.7b00017 |
| format | Article |
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Study of their molecular profiling and materials properties is currently underway for purposes of describing a variety of biological functions and diseases. However, the large, and as yet largely unquantified, variety of EV subpopulations differing in composition, size, and likely function necessitates characterization schemes capable of measuring single vesicles. Here we describe the first application of multispectral optical tweezers (MS-OTs) to single vesicles for molecular fingerprinting of EV subpopulations. This versatile imaging platform allows for sensitive measurement of Raman chemical composition (e.g., variation in protein, lipid, cholesterol, nucleic acids), coupled with discrimination by fluorescence markers. For exosomes isolated by ultracentrifugation, we use MS-OTs to interrogate the CD9-positive subpopulations via antibody fluorescence labeling and Raman spectra measurement. We report that the CD9-positive exosome subset exhibits reduced component concentration per vesicle and reduced chemical heterogeneity compared to the total purified EV population. We observed that specific vesicle subpopulations are present across exosomes isolated from cell culture supernatant of several clonal varieties of mesenchymal stromal cells and also from plasma and ascites isolated from human ovarian cancer patients.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.7b00017</identifier><identifier>PMID: 28345878</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Analytical chemistry ; Ascites ; Biochemistry ; Cancer ; CD9 antigen ; Cell culture ; Chemical composition ; Chemical fingerprinting ; Chemistry ; Cholesterol ; Coupling (molecular) ; Diseases ; Exosomes ; Fingerprinting ; Fluorescence ; Heterogeneity ; Imaging ; Labels ; Markers ; Marking ; Mesenchyme ; Nanostructure ; Nucleic acids ; Ovarian cancer ; Patients ; Plasma ; Proteins ; Raman spectra ; Raman spectroscopy ; Stromal cells ; Subpopulations ; Ultracentrifugation ; Vesicles</subject><ispartof>Analytical chemistry (Washington), 2017-05, Vol.89 (10), p.5357-5363</ispartof><rights>Copyright © 2017 American Chemical Society</rights><rights>Copyright American Chemical Society May 16, 2017</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a543t-a64c9d40340b2e968ebeb749da875e6532d81e4460f4f6da4b4a09121a6c6fa83</citedby><cites>FETCH-LOGICAL-a543t-a64c9d40340b2e968ebeb749da875e6532d81e4460f4f6da4b4a09121a6c6fa83</cites><orcidid>0000-0001-8193-1664</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/acs.analchem.7b00017$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.analchem.7b00017$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>230,314,780,784,885,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28345878$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Carney, Randy P</creatorcontrib><creatorcontrib>Hazari, Sidhartha</creatorcontrib><creatorcontrib>Colquhoun, Macalistair</creatorcontrib><creatorcontrib>Tran, Di</creatorcontrib><creatorcontrib>Hwang, Billanna</creatorcontrib><creatorcontrib>Mulligan, Michael S</creatorcontrib><creatorcontrib>Bryers, James D</creatorcontrib><creatorcontrib>Girda, Eugenia</creatorcontrib><creatorcontrib>Leiserowitz, Gary S</creatorcontrib><creatorcontrib>Smith, Zachary J</creatorcontrib><creatorcontrib>Lam, Kit S</creatorcontrib><title>Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Extracellular vesicles (EVs), including exosomes, are circulating nanoscale particles heavily implicated in cell signaling and can be isolated in vast numbers from human biofluids. Study of their molecular profiling and materials properties is currently underway for purposes of describing a variety of biological functions and diseases. However, the large, and as yet largely unquantified, variety of EV subpopulations differing in composition, size, and likely function necessitates characterization schemes capable of measuring single vesicles. Here we describe the first application of multispectral optical tweezers (MS-OTs) to single vesicles for molecular fingerprinting of EV subpopulations. This versatile imaging platform allows for sensitive measurement of Raman chemical composition (e.g., variation in protein, lipid, cholesterol, nucleic acids), coupled with discrimination by fluorescence markers. For exosomes isolated by ultracentrifugation, we use MS-OTs to interrogate the CD9-positive subpopulations via antibody fluorescence labeling and Raman spectra measurement. We report that the CD9-positive exosome subset exhibits reduced component concentration per vesicle and reduced chemical heterogeneity compared to the total purified EV population. We observed that specific vesicle subpopulations are present across exosomes isolated from cell culture supernatant of several clonal varieties of mesenchymal stromal cells and also from plasma and ascites isolated from human ovarian cancer patients.</description><subject>Analytical chemistry</subject><subject>Ascites</subject><subject>Biochemistry</subject><subject>Cancer</subject><subject>CD9 antigen</subject><subject>Cell culture</subject><subject>Chemical composition</subject><subject>Chemical fingerprinting</subject><subject>Chemistry</subject><subject>Cholesterol</subject><subject>Coupling (molecular)</subject><subject>Diseases</subject><subject>Exosomes</subject><subject>Fingerprinting</subject><subject>Fluorescence</subject><subject>Heterogeneity</subject><subject>Imaging</subject><subject>Labels</subject><subject>Markers</subject><subject>Marking</subject><subject>Mesenchyme</subject><subject>Nanostructure</subject><subject>Nucleic acids</subject><subject>Ovarian cancer</subject><subject>Patients</subject><subject>Plasma</subject><subject>Proteins</subject><subject>Raman spectra</subject><subject>Raman spectroscopy</subject><subject>Stromal cells</subject><subject>Subpopulations</subject><subject>Ultracentrifugation</subject><subject>Vesicles</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><recordid>eNp9kU1P3DAQhi1UVBbaf4CqSL1wyTJOHMe5VCrLp7QVlYCz5TiTXaMkTu2Ej_76etllKT30NJbnmXc-XkIOKUwpJPRYaT9VnWr0EttpXgIAzXfIhGYJxFyI5AOZhL80TnKAPbLv_X0gKFD-kewlImWZyMWEVD_GZjC-Rz041UTX_WB0iLePiL_R-ai2LjoxdtXkJXFuugW63pluCK_I1tHstIh_Wm8G84DR2ZP1tsXoZix724-NGozt_CeyW6vG4-dNPCB352e3s8t4fn1xNfs-j1XG0iFWnOmiYpAyKBMsuMASy5wVlRJ5hjxLk0pQZIxDzWpeKVYyBQVNqOKa10qkB-TbWrcfyxYrjd1qKRmmbZV7llYZ-T7TmaVc2AeZZRllwILA0UbA2V8j-kG2xmtsGtWhHb2kQtA8B1oUAf36D3pvRxf8CFQB4b407BQotqa0s947rLfDUJArG2WwUb7aKDc2hrIvfy-yLXr1LQCwBlblb43_p_kH06eujw</recordid><startdate>20170516</startdate><enddate>20170516</enddate><creator>Carney, Randy P</creator><creator>Hazari, Sidhartha</creator><creator>Colquhoun, Macalistair</creator><creator>Tran, Di</creator><creator>Hwang, Billanna</creator><creator>Mulligan, Michael S</creator><creator>Bryers, James D</creator><creator>Girda, Eugenia</creator><creator>Leiserowitz, Gary S</creator><creator>Smith, Zachary J</creator><creator>Lam, Kit S</creator><general>American Chemical Society</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-8193-1664</orcidid></search><sort><creationdate>20170516</creationdate><title>Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations</title><author>Carney, Randy P ; Hazari, Sidhartha ; Colquhoun, Macalistair ; Tran, Di ; Hwang, Billanna ; Mulligan, Michael S ; Bryers, James D ; Girda, Eugenia ; Leiserowitz, Gary S ; Smith, Zachary J ; Lam, Kit S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a543t-a64c9d40340b2e968ebeb749da875e6532d81e4460f4f6da4b4a09121a6c6fa83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Analytical chemistry</topic><topic>Ascites</topic><topic>Biochemistry</topic><topic>Cancer</topic><topic>CD9 antigen</topic><topic>Cell culture</topic><topic>Chemical composition</topic><topic>Chemical fingerprinting</topic><topic>Chemistry</topic><topic>Cholesterol</topic><topic>Coupling (molecular)</topic><topic>Diseases</topic><topic>Exosomes</topic><topic>Fingerprinting</topic><topic>Fluorescence</topic><topic>Heterogeneity</topic><topic>Imaging</topic><topic>Labels</topic><topic>Markers</topic><topic>Marking</topic><topic>Mesenchyme</topic><topic>Nanostructure</topic><topic>Nucleic acids</topic><topic>Ovarian cancer</topic><topic>Patients</topic><topic>Plasma</topic><topic>Proteins</topic><topic>Raman spectra</topic><topic>Raman spectroscopy</topic><topic>Stromal cells</topic><topic>Subpopulations</topic><topic>Ultracentrifugation</topic><topic>Vesicles</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Carney, Randy P</creatorcontrib><creatorcontrib>Hazari, Sidhartha</creatorcontrib><creatorcontrib>Colquhoun, Macalistair</creatorcontrib><creatorcontrib>Tran, Di</creatorcontrib><creatorcontrib>Hwang, Billanna</creatorcontrib><creatorcontrib>Mulligan, Michael S</creatorcontrib><creatorcontrib>Bryers, James D</creatorcontrib><creatorcontrib>Girda, Eugenia</creatorcontrib><creatorcontrib>Leiserowitz, Gary S</creatorcontrib><creatorcontrib>Smith, Zachary J</creatorcontrib><creatorcontrib>Lam, Kit S</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Carney, Randy P</au><au>Hazari, Sidhartha</au><au>Colquhoun, Macalistair</au><au>Tran, Di</au><au>Hwang, Billanna</au><au>Mulligan, Michael S</au><au>Bryers, James D</au><au>Girda, Eugenia</au><au>Leiserowitz, Gary S</au><au>Smith, Zachary J</au><au>Lam, Kit S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2017-05-16</date><risdate>2017</risdate><volume>89</volume><issue>10</issue><spage>5357</spage><epage>5363</epage><pages>5357-5363</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><abstract>Extracellular vesicles (EVs), including exosomes, are circulating nanoscale particles heavily implicated in cell signaling and can be isolated in vast numbers from human biofluids. Study of their molecular profiling and materials properties is currently underway for purposes of describing a variety of biological functions and diseases. However, the large, and as yet largely unquantified, variety of EV subpopulations differing in composition, size, and likely function necessitates characterization schemes capable of measuring single vesicles. Here we describe the first application of multispectral optical tweezers (MS-OTs) to single vesicles for molecular fingerprinting of EV subpopulations. This versatile imaging platform allows for sensitive measurement of Raman chemical composition (e.g., variation in protein, lipid, cholesterol, nucleic acids), coupled with discrimination by fluorescence markers. For exosomes isolated by ultracentrifugation, we use MS-OTs to interrogate the CD9-positive subpopulations via antibody fluorescence labeling and Raman spectra measurement. We report that the CD9-positive exosome subset exhibits reduced component concentration per vesicle and reduced chemical heterogeneity compared to the total purified EV population. We observed that specific vesicle subpopulations are present across exosomes isolated from cell culture supernatant of several clonal varieties of mesenchymal stromal cells and also from plasma and ascites isolated from human ovarian cancer patients.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>28345878</pmid><doi>10.1021/acs.analchem.7b00017</doi><tpages>7</tpages><orcidid>https://orcid.org/0000-0001-8193-1664</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Analytical chemistry Ascites Biochemistry Cancer CD9 antigen Cell culture Chemical composition Chemical fingerprinting Chemistry Cholesterol Coupling (molecular) Diseases Exosomes Fingerprinting Fluorescence Heterogeneity Imaging Labels Markers Marking Mesenchyme Nanostructure Nucleic acids Ovarian cancer Patients Plasma Proteins Raman spectra Raman spectroscopy Stromal cells Subpopulations Ultracentrifugation Vesicles |
| title | Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations |
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