Patch-clamping of primary cardiac cells with micro-openings in polyimide films

Abstract Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate...
Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Stett, A. [verfasserIn] Bucher, V. Burkhardt, C. Weber, U. Nisch, W.

Format:	Artikel
Sprache:	Englisch

Erschienen:	2003

Schlagwörter:	High-throughput screening Ion channel Electrophysiology Patch-clamping Loose patch Automation

Anmerkung:	© IFMBE 2003

Übergeordnetes Werk:	Enthalten in: Medical & biological engineering & computing - Springer-Verlag, 1977, 41(2003), 2 vom: März, Seite 233-240
Übergeordnetes Werk:	volume:41 ; year:2003 ; number:2 ; month:03 ; pages:233-240

Links:	Volltext

DOI / URN:	10.1007/BF02344895

Katalog-ID:	OLC2038678863

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520			\|a Abstract Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate was tested. Using standard microfabrication and ion beam technology, small-diameter openings (2 and 4 μm) were formed in polyimide films (thickness 6.5 μm). Single cells (sheep Purkinje heart cells, Chinese hamster ovary cells) in a suspension were positioned on top of the opening and sucked towards the opening to improve adhesion of the cell to the planar substrate, hence increasing the seal resistance. Voltage/current measurements yielded a median seal resistance of 1.3MΩ with 4 μm openings (n=24) and 26.0 MΩ with 2 μm openings (n=75), respectively. With 2 μm openings, successful loose-patch recordings of TTX-sensitive inward currents and action potentials in sheep Purkinje heart cells (n=18) were made. In rare cases, gigaseals (n=4) were also measured, and a whole-cell configuration (n=1) could be established. It was concluded that the simple planar patch approach is suitable for automated loosepatch recordings from cells in suspension but will hardly be suitable for highthroughput whole-cell patch-clamping with high-resistance seals.
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allfields	10.1007/BF02344895 doi (DE-627)OLC2038678863 (DE-He213)BF02344895-p DE-627 ger DE-627 rakwb eng 610 660 570 VZ 12 ssgn Stett, A. verfasserin aut Patch-clamping of primary cardiac cells with micro-openings in polyimide films 2003 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © IFMBE 2003 Abstract Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate was tested. Using standard microfabrication and ion beam technology, small-diameter openings (2 and 4 μm) were formed in polyimide films (thickness 6.5 μm). Single cells (sheep Purkinje heart cells, Chinese hamster ovary cells) in a suspension were positioned on top of the opening and sucked towards the opening to improve adhesion of the cell to the planar substrate, hence increasing the seal resistance. Voltage/current measurements yielded a median seal resistance of 1.3MΩ with 4 μm openings (n=24) and 26.0 MΩ with 2 μm openings (n=75), respectively. With 2 μm openings, successful loose-patch recordings of TTX-sensitive inward currents and action potentials in sheep Purkinje heart cells (n=18) were made. In rare cases, gigaseals (n=4) were also measured, and a whole-cell configuration (n=1) could be established. It was concluded that the simple planar patch approach is suitable for automated loosepatch recordings from cells in suspension but will hardly be suitable for highthroughput whole-cell patch-clamping with high-resistance seals. High-throughput screening Ion channel Electrophysiology Patch-clamping Loose patch Automation Bucher, V. aut Burkhardt, C. aut Weber, U. aut Nisch, W. aut Enthalten in Medical & biological engineering & computing Springer-Verlag, 1977 41(2003), 2 vom: März, Seite 233-240 (DE-627)129858552 (DE-600)282327-5 (DE-576)015165507 0140-0118 nnns volume:41 year:2003 number:2 month:03 pages:233-240 https://doi.org/10.1007/BF02344895 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 SSG-OPC-MAT GBV_ILN_32 GBV_ILN_40 GBV_ILN_70 GBV_ILN_105 GBV_ILN_118 GBV_ILN_2006 GBV_ILN_4012 GBV_ILN_4219 GBV_ILN_4306 GBV_ILN_4307 AR 41 2003 2 03 233-240
spelling	10.1007/BF02344895 doi (DE-627)OLC2038678863 (DE-He213)BF02344895-p DE-627 ger DE-627 rakwb eng 610 660 570 VZ 12 ssgn Stett, A. verfasserin aut Patch-clamping of primary cardiac cells with micro-openings in polyimide films 2003 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © IFMBE 2003 Abstract Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate was tested. Using standard microfabrication and ion beam technology, small-diameter openings (2 and 4 μm) were formed in polyimide films (thickness 6.5 μm). Single cells (sheep Purkinje heart cells, Chinese hamster ovary cells) in a suspension were positioned on top of the opening and sucked towards the opening to improve adhesion of the cell to the planar substrate, hence increasing the seal resistance. Voltage/current measurements yielded a median seal resistance of 1.3MΩ with 4 μm openings (n=24) and 26.0 MΩ with 2 μm openings (n=75), respectively. With 2 μm openings, successful loose-patch recordings of TTX-sensitive inward currents and action potentials in sheep Purkinje heart cells (n=18) were made. In rare cases, gigaseals (n=4) were also measured, and a whole-cell configuration (n=1) could be established. It was concluded that the simple planar patch approach is suitable for automated loosepatch recordings from cells in suspension but will hardly be suitable for highthroughput whole-cell patch-clamping with high-resistance seals. High-throughput screening Ion channel Electrophysiology Patch-clamping Loose patch Automation Bucher, V. aut Burkhardt, C. aut Weber, U. aut Nisch, W. aut Enthalten in Medical & biological engineering & computing Springer-Verlag, 1977 41(2003), 2 vom: März, Seite 233-240 (DE-627)129858552 (DE-600)282327-5 (DE-576)015165507 0140-0118 nnns volume:41 year:2003 number:2 month:03 pages:233-240 https://doi.org/10.1007/BF02344895 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 SSG-OPC-MAT GBV_ILN_32 GBV_ILN_40 GBV_ILN_70 GBV_ILN_105 GBV_ILN_118 GBV_ILN_2006 GBV_ILN_4012 GBV_ILN_4219 GBV_ILN_4306 GBV_ILN_4307 AR 41 2003 2 03 233-240
allfields_unstemmed	10.1007/BF02344895 doi (DE-627)OLC2038678863 (DE-He213)BF02344895-p DE-627 ger DE-627 rakwb eng 610 660 570 VZ 12 ssgn Stett, A. verfasserin aut Patch-clamping of primary cardiac cells with micro-openings in polyimide films 2003 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © IFMBE 2003 Abstract Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate was tested. Using standard microfabrication and ion beam technology, small-diameter openings (2 and 4 μm) were formed in polyimide films (thickness 6.5 μm). Single cells (sheep Purkinje heart cells, Chinese hamster ovary cells) in a suspension were positioned on top of the opening and sucked towards the opening to improve adhesion of the cell to the planar substrate, hence increasing the seal resistance. Voltage/current measurements yielded a median seal resistance of 1.3MΩ with 4 μm openings (n=24) and 26.0 MΩ with 2 μm openings (n=75), respectively. With 2 μm openings, successful loose-patch recordings of TTX-sensitive inward currents and action potentials in sheep Purkinje heart cells (n=18) were made. In rare cases, gigaseals (n=4) were also measured, and a whole-cell configuration (n=1) could be established. It was concluded that the simple planar patch approach is suitable for automated loosepatch recordings from cells in suspension but will hardly be suitable for highthroughput whole-cell patch-clamping with high-resistance seals. High-throughput screening Ion channel Electrophysiology Patch-clamping Loose patch Automation Bucher, V. aut Burkhardt, C. aut Weber, U. aut Nisch, W. aut Enthalten in Medical & biological engineering & computing Springer-Verlag, 1977 41(2003), 2 vom: März, Seite 233-240 (DE-627)129858552 (DE-600)282327-5 (DE-576)015165507 0140-0118 nnns volume:41 year:2003 number:2 month:03 pages:233-240 https://doi.org/10.1007/BF02344895 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 SSG-OPC-MAT GBV_ILN_32 GBV_ILN_40 GBV_ILN_70 GBV_ILN_105 GBV_ILN_118 GBV_ILN_2006 GBV_ILN_4012 GBV_ILN_4219 GBV_ILN_4306 GBV_ILN_4307 AR 41 2003 2 03 233-240
allfieldsGer	10.1007/BF02344895 doi (DE-627)OLC2038678863 (DE-He213)BF02344895-p DE-627 ger DE-627 rakwb eng 610 660 570 VZ 12 ssgn Stett, A. verfasserin aut Patch-clamping of primary cardiac cells with micro-openings in polyimide films 2003 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © IFMBE 2003 Abstract Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate was tested. Using standard microfabrication and ion beam technology, small-diameter openings (2 and 4 μm) were formed in polyimide films (thickness 6.5 μm). Single cells (sheep Purkinje heart cells, Chinese hamster ovary cells) in a suspension were positioned on top of the opening and sucked towards the opening to improve adhesion of the cell to the planar substrate, hence increasing the seal resistance. Voltage/current measurements yielded a median seal resistance of 1.3MΩ with 4 μm openings (n=24) and 26.0 MΩ with 2 μm openings (n=75), respectively. With 2 μm openings, successful loose-patch recordings of TTX-sensitive inward currents and action potentials in sheep Purkinje heart cells (n=18) were made. In rare cases, gigaseals (n=4) were also measured, and a whole-cell configuration (n=1) could be established. It was concluded that the simple planar patch approach is suitable for automated loosepatch recordings from cells in suspension but will hardly be suitable for highthroughput whole-cell patch-clamping with high-resistance seals. High-throughput screening Ion channel Electrophysiology Patch-clamping Loose patch Automation Bucher, V. aut Burkhardt, C. aut Weber, U. aut Nisch, W. aut Enthalten in Medical & biological engineering & computing Springer-Verlag, 1977 41(2003), 2 vom: März, Seite 233-240 (DE-627)129858552 (DE-600)282327-5 (DE-576)015165507 0140-0118 nnns volume:41 year:2003 number:2 month:03 pages:233-240 https://doi.org/10.1007/BF02344895 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 SSG-OPC-MAT GBV_ILN_32 GBV_ILN_40 GBV_ILN_70 GBV_ILN_105 GBV_ILN_118 GBV_ILN_2006 GBV_ILN_4012 GBV_ILN_4219 GBV_ILN_4306 GBV_ILN_4307 AR 41 2003 2 03 233-240
allfieldsSound	10.1007/BF02344895 doi (DE-627)OLC2038678863 (DE-He213)BF02344895-p DE-627 ger DE-627 rakwb eng 610 660 570 VZ 12 ssgn Stett, A. verfasserin aut Patch-clamping of primary cardiac cells with micro-openings in polyimide films 2003 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © IFMBE 2003 Abstract Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate was tested. Using standard microfabrication and ion beam technology, small-diameter openings (2 and 4 μm) were formed in polyimide films (thickness 6.5 μm). Single cells (sheep Purkinje heart cells, Chinese hamster ovary cells) in a suspension were positioned on top of the opening and sucked towards the opening to improve adhesion of the cell to the planar substrate, hence increasing the seal resistance. Voltage/current measurements yielded a median seal resistance of 1.3MΩ with 4 μm openings (n=24) and 26.0 MΩ with 2 μm openings (n=75), respectively. With 2 μm openings, successful loose-patch recordings of TTX-sensitive inward currents and action potentials in sheep Purkinje heart cells (n=18) were made. In rare cases, gigaseals (n=4) were also measured, and a whole-cell configuration (n=1) could be established. It was concluded that the simple planar patch approach is suitable for automated loosepatch recordings from cells in suspension but will hardly be suitable for highthroughput whole-cell patch-clamping with high-resistance seals. High-throughput screening Ion channel Electrophysiology Patch-clamping Loose patch Automation Bucher, V. aut Burkhardt, C. aut Weber, U. aut Nisch, W. aut Enthalten in Medical & biological engineering & computing Springer-Verlag, 1977 41(2003), 2 vom: März, Seite 233-240 (DE-627)129858552 (DE-600)282327-5 (DE-576)015165507 0140-0118 nnns volume:41 year:2003 number:2 month:03 pages:233-240 https://doi.org/10.1007/BF02344895 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 SSG-OPC-MAT GBV_ILN_32 GBV_ILN_40 GBV_ILN_70 GBV_ILN_105 GBV_ILN_118 GBV_ILN_2006 GBV_ILN_4012 GBV_ILN_4219 GBV_ILN_4306 GBV_ILN_4307 AR 41 2003 2 03 233-240
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topic_title	610 660 570 VZ 12 ssgn Patch-clamping of primary cardiac cells with micro-openings in polyimide films High-throughput screening Ion channel Electrophysiology Patch-clamping Loose patch Automation
topic	ddc 610 ssgn 12 misc High-throughput screening misc Ion channel misc Electrophysiology misc Patch-clamping misc Loose patch misc Automation
topic_unstemmed	ddc 610 ssgn 12 misc High-throughput screening misc Ion channel misc Electrophysiology misc Patch-clamping misc Loose patch misc Automation
topic_browse	ddc 610 ssgn 12 misc High-throughput screening misc Ion channel misc Electrophysiology misc Patch-clamping misc Loose patch misc Automation
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dewey-tens	610 - Medicine & health 660 - Chemical engineering 570 - Life sciences; biology
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title	Patch-clamping of primary cardiac cells with micro-openings in polyimide films
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title_full	Patch-clamping of primary cardiac cells with micro-openings in polyimide films
author_sort	Stett, A.
journal	Medical & biological engineering & computing
journalStr	Medical & biological engineering & computing
lang_code	eng
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author_browse	Stett, A. Bucher, V. Burkhardt, C. Weber, U. Nisch, W.
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class	610 660 570 VZ 12 ssgn
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author-letter	Stett, A.
doi_str_mv	10.1007/BF02344895
dewey-full	610 660 570
title_sort	patch-clamping of primary cardiac cells with micro-openings in polyimide films
title_auth	Patch-clamping of primary cardiac cells with micro-openings in polyimide films
abstract	Abstract Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate was tested. Using standard microfabrication and ion beam technology, small-diameter openings (2 and 4 μm) were formed in polyimide films (thickness 6.5 μm). Single cells (sheep Purkinje heart cells, Chinese hamster ovary cells) in a suspension were positioned on top of the opening and sucked towards the opening to improve adhesion of the cell to the planar substrate, hence increasing the seal resistance. Voltage/current measurements yielded a median seal resistance of 1.3MΩ with 4 μm openings (n=24) and 26.0 MΩ with 2 μm openings (n=75), respectively. With 2 μm openings, successful loose-patch recordings of TTX-sensitive inward currents and action potentials in sheep Purkinje heart cells (n=18) were made. In rare cases, gigaseals (n=4) were also measured, and a whole-cell configuration (n=1) could be established. It was concluded that the simple planar patch approach is suitable for automated loosepatch recordings from cells in suspension but will hardly be suitable for highthroughput whole-cell patch-clamping with high-resistance seals. © IFMBE 2003
abstractGer	Abstract Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate was tested. Using standard microfabrication and ion beam technology, small-diameter openings (2 and 4 μm) were formed in polyimide films (thickness 6.5 μm). Single cells (sheep Purkinje heart cells, Chinese hamster ovary cells) in a suspension were positioned on top of the opening and sucked towards the opening to improve adhesion of the cell to the planar substrate, hence increasing the seal resistance. Voltage/current measurements yielded a median seal resistance of 1.3MΩ with 4 μm openings (n=24) and 26.0 MΩ with 2 μm openings (n=75), respectively. With 2 μm openings, successful loose-patch recordings of TTX-sensitive inward currents and action potentials in sheep Purkinje heart cells (n=18) were made. In rare cases, gigaseals (n=4) were also measured, and a whole-cell configuration (n=1) could be established. It was concluded that the simple planar patch approach is suitable for automated loosepatch recordings from cells in suspension but will hardly be suitable for highthroughput whole-cell patch-clamping with high-resistance seals. © IFMBE 2003
abstract_unstemmed	Abstract Patch-clamping is a powerful method for investigating the function and regulation of ionic channels. Currently, great efforts are being made to automate this method. As a step towards this goal, the feasibility of patch-clamping primary cells with a microscopic opening in a planar substrate was tested. Using standard microfabrication and ion beam technology, small-diameter openings (2 and 4 μm) were formed in polyimide films (thickness 6.5 μm). Single cells (sheep Purkinje heart cells, Chinese hamster ovary cells) in a suspension were positioned on top of the opening and sucked towards the opening to improve adhesion of the cell to the planar substrate, hence increasing the seal resistance. Voltage/current measurements yielded a median seal resistance of 1.3MΩ with 4 μm openings (n=24) and 26.0 MΩ with 2 μm openings (n=75), respectively. With 2 μm openings, successful loose-patch recordings of TTX-sensitive inward currents and action potentials in sheep Purkinje heart cells (n=18) were made. In rare cases, gigaseals (n=4) were also measured, and a whole-cell configuration (n=1) could be established. It was concluded that the simple planar patch approach is suitable for automated loosepatch recordings from cells in suspension but will hardly be suitable for highthroughput whole-cell patch-clamping with high-resistance seals. © IFMBE 2003
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container_issue	2
title_short	Patch-clamping of primary cardiac cells with micro-openings in polyimide films
url	https://doi.org/10.1007/BF02344895
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author2	Bucher, V. Burkhardt, C. Weber, U. Nisch, W.
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up_date	2024-07-03T19:49:09.043Z
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