Spatiotemporally controlled single cell sonoporation
This paper presents unique approaches to enable control and quantification of ultrasound-mediated cell membrane disruption, or sonoporation, at the single-cell level. Ultrasound excitation of microbubbles that were targeted to the plasma membrane of HEK-293 cells generated spatially and temporally c...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2012-10, Vol.109 (41), p.16486-16491 |
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| creator | Fan, Zhenzhen Liu, Haiyan Mayer, Michael Deng, Cheri X |
| description | This paper presents unique approaches to enable control and quantification of ultrasound-mediated cell membrane disruption, or sonoporation, at the single-cell level. Ultrasound excitation of microbubbles that were targeted to the plasma membrane of HEK-293 cells generated spatially and temporally controlled membrane disruption with high repeatability. Using whole-cell patch clamp recording combined with fluorescence microscopy, we obtained time-resolved measurements of single-cell sonoporation and quantified the size and resealing rate of pores. We measured the intracellular diffusion coefficient of cytoplasmic RNA/DNA from sonoporation-induced transport of an intercalating fluorescent dye into and within single cells. We achieved spatiotemporally controlled delivery with subcellular precision and calcium signaling in targeted cells by selective excitation of microbubbles. Finally, we utilized sonoporation to deliver calcein, a membrane-impermeant substrate of multidrug resistance protein-1 (MRP1), into HEK-MRP1 cells, which overexpress MRP1, and monitored the calcein efflux by MRP1. This approach made it possible to measure the efflux rate in individual cells and to compare it directly to the efflux rate in parental control cells that do not express MRP1. |
| doi_str_mv | 10.1073/pnas.1208198109 |
| format | Article |
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Ultrasound excitation of microbubbles that were targeted to the plasma membrane of HEK-293 cells generated spatially and temporally controlled membrane disruption with high repeatability. Using whole-cell patch clamp recording combined with fluorescence microscopy, we obtained time-resolved measurements of single-cell sonoporation and quantified the size and resealing rate of pores. We measured the intracellular diffusion coefficient of cytoplasmic RNA/DNA from sonoporation-induced transport of an intercalating fluorescent dye into and within single cells. We achieved spatiotemporally controlled delivery with subcellular precision and calcium signaling in targeted cells by selective excitation of microbubbles. Finally, we utilized sonoporation to deliver calcein, a membrane-impermeant substrate of multidrug resistance protein-1 (MRP1), into HEK-MRP1 cells, which overexpress MRP1, and monitored the calcein efflux by MRP1. This approach made it possible to measure the efflux rate in individual cells and to compare it directly to the efflux rate in parental control cells that do not express MRP1.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.1208198109</identifier><identifier>PMID: 23012425</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Algorithms ; ATP-Binding Cassette, Sub-Family B, Member 1 - genetics ; ATP-Binding Cassette, Sub-Family B, Member 1 - metabolism ; Biological Sciences ; Biological Transport ; Calcium ; calcium signaling ; Cell lines ; Cell Membrane - metabolism ; Cell Membrane - physiology ; Cell Membrane Permeability - physiology ; Cell membranes ; Cells ; Cytoplasm ; Deoxyribonucleic acid ; Diffusion ; diffusivity ; DNA ; Drug resistance ; Fluoresceins - metabolism ; Fluorescence ; fluorescence microscopy ; fluorescent dyes ; HEK293 Cells ; Humans ; Imaging ; Kinetics ; Membrane Potentials - physiology ; Membranes ; Memory interference ; Microbubbles ; Microscopy, Fluorescence - methods ; Microscopy, Video - methods ; multiple drug resistance ; Nucleic Acids - metabolism ; P branes ; Patch-Clamp Techniques ; plasma membrane ; Pressure pulses ; Proteins ; Reproducibility of Results ; Ribonucleic acid ; RNA ; Single-Cell Analysis - methods ; Sonication - methods ; Time Factors ; ultrasonics ; Ultrasonics - methods ; Ultrasonography</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2012-10, Vol.109 (41), p.16486-16491</ispartof><rights>copyright © 1993-2008 National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Oct 9, 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c591t-ef1664e3a46e308afcb10ece78a63a3ae3803845b64fa0ec35155db8be182b4a3</citedby><cites>FETCH-LOGICAL-c591t-ef1664e3a46e308afcb10ece78a63a3ae3803845b64fa0ec35155db8be182b4a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/109/41.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/41763375$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/41763375$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,724,777,781,800,882,27905,27906,53772,53774,57998,58231</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23012425$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fan, Zhenzhen</creatorcontrib><creatorcontrib>Liu, Haiyan</creatorcontrib><creatorcontrib>Mayer, Michael</creatorcontrib><creatorcontrib>Deng, Cheri X</creatorcontrib><title>Spatiotemporally controlled single cell sonoporation</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>This paper presents unique approaches to enable control and quantification of ultrasound-mediated cell membrane disruption, or sonoporation, at the single-cell level. Ultrasound excitation of microbubbles that were targeted to the plasma membrane of HEK-293 cells generated spatially and temporally controlled membrane disruption with high repeatability. Using whole-cell patch clamp recording combined with fluorescence microscopy, we obtained time-resolved measurements of single-cell sonoporation and quantified the size and resealing rate of pores. We measured the intracellular diffusion coefficient of cytoplasmic RNA/DNA from sonoporation-induced transport of an intercalating fluorescent dye into and within single cells. We achieved spatiotemporally controlled delivery with subcellular precision and calcium signaling in targeted cells by selective excitation of microbubbles. Finally, we utilized sonoporation to deliver calcein, a membrane-impermeant substrate of multidrug resistance protein-1 (MRP1), into HEK-MRP1 cells, which overexpress MRP1, and monitored the calcein efflux by MRP1. This approach made it possible to measure the efflux rate in individual cells and to compare it directly to the efflux rate in parental control cells that do not express MRP1.</description><subject>Algorithms</subject><subject>ATP-Binding Cassette, Sub-Family B, Member 1 - genetics</subject><subject>ATP-Binding Cassette, Sub-Family B, Member 1 - metabolism</subject><subject>Biological Sciences</subject><subject>Biological Transport</subject><subject>Calcium</subject><subject>calcium signaling</subject><subject>Cell lines</subject><subject>Cell Membrane - metabolism</subject><subject>Cell Membrane - physiology</subject><subject>Cell Membrane Permeability - physiology</subject><subject>Cell membranes</subject><subject>Cells</subject><subject>Cytoplasm</subject><subject>Deoxyribonucleic acid</subject><subject>Diffusion</subject><subject>diffusivity</subject><subject>DNA</subject><subject>Drug resistance</subject><subject>Fluoresceins - metabolism</subject><subject>Fluorescence</subject><subject>fluorescence microscopy</subject><subject>fluorescent dyes</subject><subject>HEK293 Cells</subject><subject>Humans</subject><subject>Imaging</subject><subject>Kinetics</subject><subject>Membrane Potentials - physiology</subject><subject>Membranes</subject><subject>Memory interference</subject><subject>Microbubbles</subject><subject>Microscopy, Fluorescence - methods</subject><subject>Microscopy, Video - methods</subject><subject>multiple drug resistance</subject><subject>Nucleic Acids - metabolism</subject><subject>P branes</subject><subject>Patch-Clamp Techniques</subject><subject>plasma membrane</subject><subject>Pressure pulses</subject><subject>Proteins</subject><subject>Reproducibility of Results</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>Single-Cell Analysis - methods</subject><subject>Sonication - methods</subject><subject>Time Factors</subject><subject>ultrasonics</subject><subject>Ultrasonics - methods</subject><subject>Ultrasonography</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0U1v3CAQBmBUtWq2ac85NbXUSy9OZgBjuESqon5JkXpIckbYO956xRoHvJHy74u7m03aSy9wmIeXgWHsBOEMoRbn4-DSGXLQaDSCecEWecVSSQMv2QKA16WWXB6xNymtAcBUGl6zIy4AueTVgsnr0U19mGgzhui8fyjaMEwxeE_LIvXDylPRkvdFCkOYScbDW_aqcz7Ru_1-zG6_frm5_F5e_fz24_LzVdlWBqeSOlRKknBSkQDturZBoJZq7ZRwwpHQILSsGiU7lwuiwqpaNroh1LyRThyzi13uuG02tGwpd-a8HWO_cfHBBtfbvytD_8uuwr0VstYKRQ74tA-I4W5LabKbPs3PcQOFbbKYG0CpINv_0vnvMAfzTD_-Q9dhG4f8E3-UUtqYKqvznWpjSClSd-gbwc7Ds_Pw7NPw8onT5889-MdpZVDswXzyKc5YiRaV1CqT9zuyTlOIByOxVkLUc8SHXb1zwbpV7JO9veaACvIdxnAUvwENxbL0</recordid><startdate>20121009</startdate><enddate>20121009</enddate><creator>Fan, Zhenzhen</creator><creator>Liu, Haiyan</creator><creator>Mayer, Michael</creator><creator>Deng, Cheri X</creator><general>National Academy of Sciences</general><general>National Acad Sciences</general><scope>FBQ</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>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20121009</creationdate><title>Spatiotemporally controlled single cell sonoporation</title><author>Fan, Zhenzhen ; Liu, Haiyan ; Mayer, Michael ; Deng, Cheri X</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c591t-ef1664e3a46e308afcb10ece78a63a3ae3803845b64fa0ec35155db8be182b4a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Algorithms</topic><topic>ATP-Binding Cassette, Sub-Family B, Member 1 - genetics</topic><topic>ATP-Binding Cassette, Sub-Family B, Member 1 - metabolism</topic><topic>Biological Sciences</topic><topic>Biological Transport</topic><topic>Calcium</topic><topic>calcium signaling</topic><topic>Cell lines</topic><topic>Cell Membrane - metabolism</topic><topic>Cell Membrane - physiology</topic><topic>Cell Membrane Permeability - physiology</topic><topic>Cell membranes</topic><topic>Cells</topic><topic>Cytoplasm</topic><topic>Deoxyribonucleic acid</topic><topic>Diffusion</topic><topic>diffusivity</topic><topic>DNA</topic><topic>Drug resistance</topic><topic>Fluoresceins - metabolism</topic><topic>Fluorescence</topic><topic>fluorescence microscopy</topic><topic>fluorescent dyes</topic><topic>HEK293 Cells</topic><topic>Humans</topic><topic>Imaging</topic><topic>Kinetics</topic><topic>Membrane Potentials - physiology</topic><topic>Membranes</topic><topic>Memory interference</topic><topic>Microbubbles</topic><topic>Microscopy, Fluorescence - methods</topic><topic>Microscopy, Video - methods</topic><topic>multiple drug resistance</topic><topic>Nucleic Acids - metabolism</topic><topic>P branes</topic><topic>Patch-Clamp Techniques</topic><topic>plasma membrane</topic><topic>Pressure pulses</topic><topic>Proteins</topic><topic>Reproducibility of Results</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>Single-Cell Analysis - methods</topic><topic>Sonication - methods</topic><topic>Time Factors</topic><topic>ultrasonics</topic><topic>Ultrasonics - methods</topic><topic>Ultrasonography</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Fan, Zhenzhen</creatorcontrib><creatorcontrib>Liu, Haiyan</creatorcontrib><creatorcontrib>Mayer, Michael</creatorcontrib><creatorcontrib>Deng, Cheri X</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Fan, Zhenzhen</au><au>Liu, Haiyan</au><au>Mayer, Michael</au><au>Deng, Cheri X</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Spatiotemporally controlled single cell sonoporation</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>2012-10-09</date><risdate>2012</risdate><volume>109</volume><issue>41</issue><spage>16486</spage><epage>16491</epage><pages>16486-16491</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>This paper presents unique approaches to enable control and quantification of ultrasound-mediated cell membrane disruption, or sonoporation, at the single-cell level. Ultrasound excitation of microbubbles that were targeted to the plasma membrane of HEK-293 cells generated spatially and temporally controlled membrane disruption with high repeatability. Using whole-cell patch clamp recording combined with fluorescence microscopy, we obtained time-resolved measurements of single-cell sonoporation and quantified the size and resealing rate of pores. We measured the intracellular diffusion coefficient of cytoplasmic RNA/DNA from sonoporation-induced transport of an intercalating fluorescent dye into and within single cells. We achieved spatiotemporally controlled delivery with subcellular precision and calcium signaling in targeted cells by selective excitation of microbubbles. Finally, we utilized sonoporation to deliver calcein, a membrane-impermeant substrate of multidrug resistance protein-1 (MRP1), into HEK-MRP1 cells, which overexpress MRP1, and monitored the calcein efflux by MRP1. This approach made it possible to measure the efflux rate in individual cells and to compare it directly to the efflux rate in parental control cells that do not express MRP1.</abstract><cop>United States</cop><pub>National Academy of Sciences</pub><pmid>23012425</pmid><doi>10.1073/pnas.1208198109</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Algorithms ATP-Binding Cassette, Sub-Family B, Member 1 - genetics ATP-Binding Cassette, Sub-Family B, Member 1 - metabolism Biological Sciences Biological Transport Calcium calcium signaling Cell lines Cell Membrane - metabolism Cell Membrane - physiology Cell Membrane Permeability - physiology Cell membranes Cells Cytoplasm Deoxyribonucleic acid Diffusion diffusivity DNA Drug resistance Fluoresceins - metabolism Fluorescence fluorescence microscopy fluorescent dyes HEK293 Cells Humans Imaging Kinetics Membrane Potentials - physiology Membranes Memory interference Microbubbles Microscopy, Fluorescence - methods Microscopy, Video - methods multiple drug resistance Nucleic Acids - metabolism P branes Patch-Clamp Techniques plasma membrane Pressure pulses Proteins Reproducibility of Results Ribonucleic acid RNA Single-Cell Analysis - methods Sonication - methods Time Factors ultrasonics Ultrasonics - methods Ultrasonography |
| title | Spatiotemporally controlled single cell sonoporation |
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