Microfluidic, marker-free isolation of circulating tumor cells from blood samples
The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen–independen...
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| Veröffentlicht in: | Nature protocols 2014-03, Vol.9 (3), p.694-710 |
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| creator | Karabacak, Nezihi Murat Spuhler, Philipp S Fachin, Fabio Lim, Eugene J Pai, Vincent Ozkumur, Emre Martel, Joseph M Kojic, Nikola Smith, Kyle Chen, Pin-i Yang, Jennifer Hwang, Henry Morgan, Bailey Trautwein, Julie Barber, Thomas A Stott, Shannon L Maheswaran, Shyamala Kapur, Ravi Haber, Daniel A Toner, Mehmet |
| description | The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen–independent microfluidic CTC-iChip technology. The CTC-iChip uses deterministic lateral displacement, inertial focusing and magnetophoresis to sort up to 10
7
cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2–5 d. |
| doi_str_mv | 10.1038/nprot.2014.044 |
| format | Article |
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7
cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2–5 d.</description><identifier>ISSN: 1754-2189</identifier><identifier>EISSN: 1750-2799</identifier><identifier>DOI: 10.1038/nprot.2014.044</identifier><identifier>PMID: 24577360</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>631/1647/2230 ; 631/1647/350/877 ; 631/67/2322 ; Analysis ; Analytical Chemistry ; Antibodies ; Biological Techniques ; Blood ; Cancer ; Cell Separation - methods ; Cells ; Computational Biology/Bioinformatics ; Humans ; Insect Proteins ; Leukocytes ; Life Sciences ; Magnets ; Metastasis ; Microarrays ; Microfluidic Analytical Techniques - methods ; Neoplastic Cells, Circulating ; Organic Chemistry ; Physiological aspects ; protocol ; Sample preparation ; Tumor antigens ; Tumors</subject><ispartof>Nature protocols, 2014-03, Vol.9 (3), p.694-710</ispartof><rights>Springer Nature Limited 2013</rights><rights>COPYRIGHT 2014 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Mar 2014</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c625t-467ac992332f0fb09938a8bd096ca5571aff712b7efa050d5b53c8a0cfa484f63</citedby><cites>FETCH-LOGICAL-c625t-467ac992332f0fb09938a8bd096ca5571aff712b7efa050d5b53c8a0cfa484f63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nprot.2014.044$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nprot.2014.044$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>230,314,777,781,882,27905,27906,41469,42538,51300</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24577360$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Karabacak, Nezihi Murat</creatorcontrib><creatorcontrib>Spuhler, Philipp S</creatorcontrib><creatorcontrib>Fachin, Fabio</creatorcontrib><creatorcontrib>Lim, Eugene J</creatorcontrib><creatorcontrib>Pai, Vincent</creatorcontrib><creatorcontrib>Ozkumur, Emre</creatorcontrib><creatorcontrib>Martel, Joseph M</creatorcontrib><creatorcontrib>Kojic, Nikola</creatorcontrib><creatorcontrib>Smith, Kyle</creatorcontrib><creatorcontrib>Chen, Pin-i</creatorcontrib><creatorcontrib>Yang, Jennifer</creatorcontrib><creatorcontrib>Hwang, Henry</creatorcontrib><creatorcontrib>Morgan, Bailey</creatorcontrib><creatorcontrib>Trautwein, Julie</creatorcontrib><creatorcontrib>Barber, Thomas A</creatorcontrib><creatorcontrib>Stott, Shannon L</creatorcontrib><creatorcontrib>Maheswaran, Shyamala</creatorcontrib><creatorcontrib>Kapur, Ravi</creatorcontrib><creatorcontrib>Haber, Daniel A</creatorcontrib><creatorcontrib>Toner, Mehmet</creatorcontrib><title>Microfluidic, marker-free isolation of circulating tumor cells from blood samples</title><title>Nature protocols</title><addtitle>Nat Protoc</addtitle><addtitle>Nat Protoc</addtitle><description>The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen–independent microfluidic CTC-iChip technology. The CTC-iChip uses deterministic lateral displacement, inertial focusing and magnetophoresis to sort up to 10
7
cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2–5 d.</description><subject>631/1647/2230</subject><subject>631/1647/350/877</subject><subject>631/67/2322</subject><subject>Analysis</subject><subject>Analytical Chemistry</subject><subject>Antibodies</subject><subject>Biological Techniques</subject><subject>Blood</subject><subject>Cancer</subject><subject>Cell Separation - methods</subject><subject>Cells</subject><subject>Computational Biology/Bioinformatics</subject><subject>Humans</subject><subject>Insect Proteins</subject><subject>Leukocytes</subject><subject>Life Sciences</subject><subject>Magnets</subject><subject>Metastasis</subject><subject>Microarrays</subject><subject>Microfluidic Analytical Techniques - methods</subject><subject>Neoplastic Cells, Circulating</subject><subject>Organic Chemistry</subject><subject>Physiological aspects</subject><subject>protocol</subject><subject>Sample preparation</subject><subject>Tumor antigens</subject><subject>Tumors</subject><issn>1754-2189</issn><issn>1750-2799</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNptkk1vFSEUhonR2FrdujQkbjRxboGBATYmTeNHkxrj15owDIxUBm5hxui_l2lr7TU3LDhwnvOSc3gBeIrRBqNWHMdtTvOGIEw3iNJ74BBzhhrCpbx_FdOGYCEPwKNSLhCivO34Q3BAKOM1RIfg0wdvcnJh8YM3r-Ck8w-bG5ethb6koGefIkwOGp_Nsh7jCOdlShkaG0KBLqcJ9iGlARY9bYMtj8EDp0OxT272I_Dt7Zuvp--b84_vzk5PzhvTETY3tOPaSEnaljjkeiRlK7ToByQ7oxnjWDvHMem5dRoxNLCetUZoZJymgrquPQKvr3W3Sz_Zwdg4Zx3UNvvaxG-VtFe7mei_qzH9VBRzSRitAi9uBHK6XGyZ1eTL2pWONi1FYYYoppJyWdHn_6EXacmxtrdShPNOIP6PGnWwykeX6rtmFVUnbSck41KgSm32UHUNdvImRet8vd8peLlTUJnZ_ppHvZSizr583iteP7WUbN3tPDBSq2HUlWHUahhVDVMLnt2d4i3-1yEVOL4GSk3F0eY7re-X_AOlA8uy</recordid><startdate>20140301</startdate><enddate>20140301</enddate><creator>Karabacak, Nezihi Murat</creator><creator>Spuhler, Philipp S</creator><creator>Fachin, Fabio</creator><creator>Lim, Eugene J</creator><creator>Pai, Vincent</creator><creator>Ozkumur, Emre</creator><creator>Martel, Joseph M</creator><creator>Kojic, Nikola</creator><creator>Smith, Kyle</creator><creator>Chen, Pin-i</creator><creator>Yang, Jennifer</creator><creator>Hwang, Henry</creator><creator>Morgan, Bailey</creator><creator>Trautwein, Julie</creator><creator>Barber, Thomas A</creator><creator>Stott, Shannon L</creator><creator>Maheswaran, Shyamala</creator><creator>Kapur, Ravi</creator><creator>Haber, Daniel A</creator><creator>Toner, Mehmet</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><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>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7T5</scope><scope>7T7</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PATMY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20140301</creationdate><title>Microfluidic, marker-free isolation of circulating tumor cells from blood samples</title><author>Karabacak, Nezihi Murat ; Spuhler, Philipp S ; Fachin, Fabio ; Lim, Eugene J ; Pai, Vincent ; Ozkumur, Emre ; Martel, Joseph M ; Kojic, Nikola ; Smith, Kyle ; Chen, Pin-i ; Yang, Jennifer ; Hwang, Henry ; Morgan, Bailey ; Trautwein, Julie ; Barber, Thomas A ; Stott, Shannon L ; Maheswaran, Shyamala ; Kapur, Ravi ; Haber, Daniel A ; Toner, Mehmet</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c625t-467ac992332f0fb09938a8bd096ca5571aff712b7efa050d5b53c8a0cfa484f63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>631/1647/2230</topic><topic>631/1647/350/877</topic><topic>631/67/2322</topic><topic>Analysis</topic><topic>Analytical Chemistry</topic><topic>Antibodies</topic><topic>Biological Techniques</topic><topic>Blood</topic><topic>Cancer</topic><topic>Cell Separation - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Nature protocols</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Karabacak, Nezihi Murat</au><au>Spuhler, Philipp S</au><au>Fachin, Fabio</au><au>Lim, Eugene J</au><au>Pai, Vincent</au><au>Ozkumur, Emre</au><au>Martel, Joseph M</au><au>Kojic, Nikola</au><au>Smith, Kyle</au><au>Chen, Pin-i</au><au>Yang, Jennifer</au><au>Hwang, Henry</au><au>Morgan, Bailey</au><au>Trautwein, Julie</au><au>Barber, Thomas A</au><au>Stott, Shannon L</au><au>Maheswaran, Shyamala</au><au>Kapur, Ravi</au><au>Haber, Daniel A</au><au>Toner, Mehmet</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Microfluidic, marker-free isolation of circulating tumor cells from blood samples</atitle><jtitle>Nature protocols</jtitle><stitle>Nat Protoc</stitle><addtitle>Nat Protoc</addtitle><date>2014-03-01</date><risdate>2014</risdate><volume>9</volume><issue>3</issue><spage>694</spage><epage>710</epage><pages>694-710</pages><issn>1754-2189</issn><eissn>1750-2799</eissn><abstract>The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen–independent microfluidic CTC-iChip technology. The CTC-iChip uses deterministic lateral displacement, inertial focusing and magnetophoresis to sort up to 10
7
cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2–5 d.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>24577360</pmid><doi>10.1038/nprot.2014.044</doi><tpages>17</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | 631/1647/2230 631/1647/350/877 631/67/2322 Analysis Analytical Chemistry Antibodies Biological Techniques Blood Cancer Cell Separation - methods Cells Computational Biology/Bioinformatics Humans Insect Proteins Leukocytes Life Sciences Magnets Metastasis Microarrays Microfluidic Analytical Techniques - methods Neoplastic Cells, Circulating Organic Chemistry Physiological aspects protocol Sample preparation Tumor antigens Tumors |
| title | Microfluidic, marker-free isolation of circulating tumor cells from blood samples |
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