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Microfluidic-Based Cell Sorting of Francisella tularensis Infected Macrophages Using Optical Forces
We have extended the principle of optical tweezers as a noninvasive technique to actively sort hydrodynamically focused cells based on their fluorescence signal in a microfluidic device. This micro fluorescence-activated cell sorter (µFACS) uses an infrared laser to laterally deflect cells into a co...
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| Published in: | Analytical chemistry (Washington) 2008-08, Vol.80 (16), p.6365-6372 |
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| Main Authors: | , , , , , , |
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| cited_by | cdi_FETCH-LOGICAL-a532t-50e616fe9206cabc78329616c1ff0551f545b8170460ca36fdfa2fe28a706b353 |
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| container_end_page | 6372 |
| container_issue | 16 |
| container_start_page | 6365 |
| container_title | Analytical chemistry (Washington) |
| container_volume | 80 |
| creator | Perroud, Thomas D Kaiser, Julia N Sy, Jay C Lane, Todd W Branda, Catherine S Singh, Anup K Patel, Kamlesh D |
| description | We have extended the principle of optical tweezers as a noninvasive technique to actively sort hydrodynamically focused cells based on their fluorescence signal in a microfluidic device. This micro fluorescence-activated cell sorter (µFACS) uses an infrared laser to laterally deflect cells into a collection channel. Green-labeled macrophages were sorted from a 40/60 ratio mixture at a throughput of 22 cells/s over 30 min achieving a 93% sorting purity and a 60% recovery yield. To rule out potential photoinduced cell damage during optical deflection, we investigated the response of mouse macrophage to brief exposures ( |
| doi_str_mv | 10.1021/ac8007779 |
| format | article |
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(SNL-CA), Livermore, CA (United States)</creatorcontrib><description>We have extended the principle of optical tweezers as a noninvasive technique to actively sort hydrodynamically focused cells based on their fluorescence signal in a microfluidic device. This micro fluorescence-activated cell sorter (µFACS) uses an infrared laser to laterally deflect cells into a collection channel. Green-labeled macrophages were sorted from a 40/60 ratio mixture at a throughput of 22 cells/s over 30 min achieving a 93% sorting purity and a 60% recovery yield. To rule out potential photoinduced cell damage during optical deflection, we investigated the response of mouse macrophage to brief exposures (<4 ms) of focused 1064-nm laser light (9.6 W at the sample). We found no significant difference in viability, cell proliferation, activation state, and functionality between infrared-exposed and unexposed cells. Activation state was measured by the phosphorylation of ERK and nuclear translocation of NF-κB, while functionality was assessed in a similar manner, but after a lipopolysaccharide challenge. To demonstrate the selective nature of optical sorting, we isolated a subpopulation of macrophages highly infected with the fluorescently labeled pathogen Francisella tularensis subsp. novicida. A total of 10 738 infected cells were sorted at a throughput of 11 cells/s with 93% purity and 39% recovery.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/ac8007779</identifier><identifier>PMID: 18510341</identifier><identifier>CODEN: ANCHAM</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Analytical chemistry ; Animals ; Bacteria ; Cell Nucleus - metabolism ; Cell Proliferation ; Cell Separation - methods ; Cell Survival ; Cells ; Cells, Cultured ; Chemistry ; Diagnostic Imaging ; Exact sciences and technology ; Extracellular Signal-Regulated MAP Kinases - metabolism ; Flow Cytometry ; Fluorescence ; Fluorescent Dyes ; Francisella tularensis ; Francisella tularensis - immunology ; Francisella tularensis - metabolism ; Francisella tularensis - radiation effects ; Green Fluorescent Proteins ; Infrared radiation ; Lipopolysaccharides - pharmacology ; Macrophage Activation - radiation effects ; Macrophages - immunology ; Macrophages - microbiology ; Macrophages - radiation effects ; Mice ; Microfluidics - methods ; NF-kappa B - metabolism ; Optical Tweezers ; Optics ; Phosphorylation - radiation effects ; Protein Transport ; Signal Transduction ; Spectrometric and optical methods ; Tularemia - immunology</subject><ispartof>Analytical chemistry (Washington), 2008-08, Vol.80 (16), p.6365-6372</ispartof><rights>Copyright © 2008 American Chemical Society</rights><rights>2008 INIST-CNRS</rights><rights>Copyright American Chemical Society Aug 15, 2008</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a532t-50e616fe9206cabc78329616c1ff0551f545b8170460ca36fdfa2fe28a706b353</citedby><cites>FETCH-LOGICAL-a532t-50e616fe9206cabc78329616c1ff0551f545b8170460ca36fdfa2fe28a706b353</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,777,781,882,27906,27907</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=20606434$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18510341$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/1145788$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Perroud, Thomas D</creatorcontrib><creatorcontrib>Kaiser, Julia N</creatorcontrib><creatorcontrib>Sy, Jay C</creatorcontrib><creatorcontrib>Lane, Todd W</creatorcontrib><creatorcontrib>Branda, Catherine S</creatorcontrib><creatorcontrib>Singh, Anup K</creatorcontrib><creatorcontrib>Patel, Kamlesh D</creatorcontrib><creatorcontrib>Sandia National Lab. (SNL-CA), Livermore, CA (United States)</creatorcontrib><title>Microfluidic-Based Cell Sorting of Francisella tularensis Infected Macrophages Using Optical Forces</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>We have extended the principle of optical tweezers as a noninvasive technique to actively sort hydrodynamically focused cells based on their fluorescence signal in a microfluidic device. This micro fluorescence-activated cell sorter (µFACS) uses an infrared laser to laterally deflect cells into a collection channel. Green-labeled macrophages were sorted from a 40/60 ratio mixture at a throughput of 22 cells/s over 30 min achieving a 93% sorting purity and a 60% recovery yield. To rule out potential photoinduced cell damage during optical deflection, we investigated the response of mouse macrophage to brief exposures (<4 ms) of focused 1064-nm laser light (9.6 W at the sample). We found no significant difference in viability, cell proliferation, activation state, and functionality between infrared-exposed and unexposed cells. Activation state was measured by the phosphorylation of ERK and nuclear translocation of NF-κB, while functionality was assessed in a similar manner, but after a lipopolysaccharide challenge. To demonstrate the selective nature of optical sorting, we isolated a subpopulation of macrophages highly infected with the fluorescently labeled pathogen Francisella tularensis subsp. novicida. A total of 10 738 infected cells were sorted at a throughput of 11 cells/s with 93% purity and 39% recovery.</description><subject>Analytical chemistry</subject><subject>Animals</subject><subject>Bacteria</subject><subject>Cell Nucleus - metabolism</subject><subject>Cell Proliferation</subject><subject>Cell Separation - methods</subject><subject>Cell Survival</subject><subject>Cells</subject><subject>Cells, Cultured</subject><subject>Chemistry</subject><subject>Diagnostic Imaging</subject><subject>Exact sciences and technology</subject><subject>Extracellular Signal-Regulated MAP Kinases - metabolism</subject><subject>Flow Cytometry</subject><subject>Fluorescence</subject><subject>Fluorescent Dyes</subject><subject>Francisella tularensis</subject><subject>Francisella tularensis - immunology</subject><subject>Francisella tularensis - metabolism</subject><subject>Francisella tularensis - radiation effects</subject><subject>Green Fluorescent Proteins</subject><subject>Infrared radiation</subject><subject>Lipopolysaccharides - pharmacology</subject><subject>Macrophage Activation - radiation effects</subject><subject>Macrophages - immunology</subject><subject>Macrophages - microbiology</subject><subject>Macrophages - radiation effects</subject><subject>Mice</subject><subject>Microfluidics - methods</subject><subject>NF-kappa B - metabolism</subject><subject>Optical Tweezers</subject><subject>Optics</subject><subject>Phosphorylation - radiation effects</subject><subject>Protein Transport</subject><subject>Signal Transduction</subject><subject>Spectrometric and optical methods</subject><subject>Tularemia - immunology</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><recordid>eNqFkUFvEzEQhS0EomnhwB9AKySQOCzM2Gt79wgRoZVaNSitKnGxHMduXTa7wbMrlX-Po0TJgQMnS-Nv3sybx9gbhE8IHD9bVwNorZtnbIKSQ6nqmj9nEwAQJdcAJ-yU6BEAEVC9ZCdYSwRR4YS5q-hSH9oxrqIrv1ryq2Lq27ZY9GmI3X3Rh2KWbOci5aothrG1yXcUqbjogndD5q9sltg82HtPxS1tm643Q3S2LWZ9cp5esRfBtuRf798zdjv7djM9Ly-vv19Mv1yWVgo-lBK8QhV8w0E5u3S6FrzJFYchgJQYZCWXNWqoFDgrVFgFy4PntdWglkKKM_Zup9vTEA25OHj34Pquy2saxErqus7Qhx20Sf3v0dNg1pHc1lvn-5GMairBK4n_BbFRkleNPo49gI_9mLrs1HDUDYKqVIY-7qB8KqLkg9mkuLbpj0Ew2xDNIcTMvt0Ljsu1Xx3JfWoZeL8HLOUzh10-By7fL88UVebKHRdp8E-Hf5t-GaWFluZmvjA_fp7P79Ribu6OutbR0cS_C_4FI4q81Q</recordid><startdate>20080815</startdate><enddate>20080815</enddate><creator>Perroud, Thomas D</creator><creator>Kaiser, Julia N</creator><creator>Sy, Jay C</creator><creator>Lane, Todd W</creator><creator>Branda, Catherine S</creator><creator>Singh, Anup K</creator><creator>Patel, Kamlesh D</creator><general>American Chemical Society</general><scope>BSCLL</scope><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>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>7QL</scope><scope>7X8</scope><scope>OTOTI</scope></search><sort><creationdate>20080815</creationdate><title>Microfluidic-Based Cell Sorting of Francisella tularensis Infected Macrophages Using Optical Forces</title><author>Perroud, Thomas D ; 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(SNL-CA), Livermore, CA (United States)</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Microfluidic-Based Cell Sorting of Francisella tularensis Infected Macrophages Using Optical Forces</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2008-08-15</date><risdate>2008</risdate><volume>80</volume><issue>16</issue><spage>6365</spage><epage>6372</epage><pages>6365-6372</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>We have extended the principle of optical tweezers as a noninvasive technique to actively sort hydrodynamically focused cells based on their fluorescence signal in a microfluidic device. This micro fluorescence-activated cell sorter (µFACS) uses an infrared laser to laterally deflect cells into a collection channel. Green-labeled macrophages were sorted from a 40/60 ratio mixture at a throughput of 22 cells/s over 30 min achieving a 93% sorting purity and a 60% recovery yield. To rule out potential photoinduced cell damage during optical deflection, we investigated the response of mouse macrophage to brief exposures (<4 ms) of focused 1064-nm laser light (9.6 W at the sample). We found no significant difference in viability, cell proliferation, activation state, and functionality between infrared-exposed and unexposed cells. Activation state was measured by the phosphorylation of ERK and nuclear translocation of NF-κB, while functionality was assessed in a similar manner, but after a lipopolysaccharide challenge. To demonstrate the selective nature of optical sorting, we isolated a subpopulation of macrophages highly infected with the fluorescently labeled pathogen Francisella tularensis subsp. novicida. A total of 10 738 infected cells were sorted at a throughput of 11 cells/s with 93% purity and 39% recovery.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>18510341</pmid><doi>10.1021/ac8007779</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Analytical chemistry (Washington), 2008-08, Vol.80 (16), p.6365-6372 |
| issn | 0003-2700 1520-6882 |
| language | eng |
| recordid | cdi_osti_scitechconnect_1145788 |
| source | American Chemical Society:Jisc Collections:American Chemical Society Read & Publish Agreement 2022-2024 (Reading list) |
| subjects | Analytical chemistry Animals Bacteria Cell Nucleus - metabolism Cell Proliferation Cell Separation - methods Cell Survival Cells Cells, Cultured Chemistry Diagnostic Imaging Exact sciences and technology Extracellular Signal-Regulated MAP Kinases - metabolism Flow Cytometry Fluorescence Fluorescent Dyes Francisella tularensis Francisella tularensis - immunology Francisella tularensis - metabolism Francisella tularensis - radiation effects Green Fluorescent Proteins Infrared radiation Lipopolysaccharides - pharmacology Macrophage Activation - radiation effects Macrophages - immunology Macrophages - microbiology Macrophages - radiation effects Mice Microfluidics - methods NF-kappa B - metabolism Optical Tweezers Optics Phosphorylation - radiation effects Protein Transport Signal Transduction Spectrometric and optical methods Tularemia - immunology |
| title | Microfluidic-Based Cell Sorting of Francisella tularensis Infected Macrophages Using Optical Forces |
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