Ratiometry of transmembrane voltage-sensitive fluorescent dye emission in hearts
Department of Biomedical Engineering of the School of Engineering, The University of Alabama at Birmingham, Alabama 35294-0019 Transmembrane voltage-sensitive fluorescence measurements are limited by baseline drift that can obscure changes in resting membrane potential and by motion artifacts that c...
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| Veröffentlicht in: | American journal of physiology. Heart and circulatory physiology 2000-09, Vol.279 (3), p.H1421-H1433 |
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| container_title | American journal of physiology. Heart and circulatory physiology |
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| creator | Knisley, Stephen B Justice, Robert K Kong, Wei Johnson, Philip L |
| description | Department of Biomedical Engineering of the School of
Engineering, The University of Alabama at Birmingham, Alabama
35294-0019
Transmembrane voltage-sensitive
fluorescence measurements are limited by baseline drift that can
obscure changes in resting membrane potential and by motion artifacts
that can obscure repolarization. Voltage-dependent shift of emission
wavelengths may allow reduction of drift and motion artifacts by
emission ratiometry. We have tested this for action potentials and
potassium-induced changes in resting membrane potential in rabbit
hearts stained with di-4-ANEPPS [Pyridinium,
4-(2-(6-(dibutylamino)-2-naphthalenyl) ethenyl)-1-(3-sulfopropyl)-, hydroxide, inner salt] using laser excitation (488 nm) and a
two-photomultiplier tube system or spectrofluorometer (resolution of
500-1,000 Hz and |
| doi_str_mv | 10.1152/ajpheart.2000.279.3.h1421 |
| format | Article |
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Engineering, The University of Alabama at Birmingham, Alabama
35294-0019
Transmembrane voltage-sensitive
fluorescence measurements are limited by baseline drift that can
obscure changes in resting membrane potential and by motion artifacts
that can obscure repolarization. Voltage-dependent shift of emission
wavelengths may allow reduction of drift and motion artifacts by
emission ratiometry. We have tested this for action potentials and
potassium-induced changes in resting membrane potential in rabbit
hearts stained with di-4-ANEPPS [Pyridinium,
4-(2-(6-(dibutylamino)-2-naphthalenyl) ethenyl)-1-(3-sulfopropyl)-, hydroxide, inner salt] using laser excitation (488 nm) and a
two-photomultiplier tube system or spectrofluorometer (resolution of
500-1,000 Hz and <1 mm). Green and red emissions produced upright
and inverted action potentials, respectively. Ratios of green emission
to red emission followed action potential contours and exhibited larger fractional changes than either emission alone ( P < 0.001). The largest changes and signal-to-noise ratio (signal/noise)
were obtained with numerator wavelengths of 525-550 nm and
denominator wavelengths of 650-700 nm. Ratiometry lessened drift
56-66% ( P < 0.015) and indicated decreases in
resting membrane potential. Ratiometry lessened motion
artifacts and increased magnitudes of deflections representing
phase-zero depolarizations relative to total deflections by
123-188% in intact hearts ( P < 0.02). Durations
of action potentials at different pacing rates, temperatures, and
potassium concentrations were independent of whether they were measured
ratiometrically or with microelectrodes ( P 0.65). The
ratiometric calibration slope was 0.017/100 mV and decreased with time.
Thus emission ratiometry lessens the effects of motion and drift and
indicates resting membrane potential changes and repolarization.
potentiometric dye; Pyridinium,
4-(2-(6-(dibutylamino)-2-naphthalenyl) ethenyl)-1-(3-sulfopropyl)-,
hydroxide, inner salt; potassium; action potential</description><identifier>ISSN: 0363-6135</identifier><identifier>EISSN: 1522-1539</identifier><identifier>DOI: 10.1152/ajpheart.2000.279.3.h1421</identifier><identifier>PMID: 10993810</identifier><language>eng</language><publisher>United States</publisher><subject>Animals ; Body Surface Potential Mapping ; Cardiac Pacing, Artificial ; Evaluation Studies as Topic ; Fluorescent Dyes ; In Vitro Techniques ; Lasers ; Membrane Potentials - drug effects ; Membrane Potentials - physiology ; Microelectrodes ; Myocardium - metabolism ; Potassium - metabolism ; Potassium - pharmacology ; Potentiometry - methods ; Pyridinium Compounds ; Rabbits ; Spectrometry, Fluorescence - instrumentation ; Spectrometry, Fluorescence - methods ; Temperature</subject><ispartof>American journal of physiology. Heart and circulatory physiology, 2000-09, Vol.279 (3), p.H1421-H1433</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c471t-fdc37205164ece81709889add3ba302d06cbe881e72c849dd59e1cafb84f69c93</citedby><cites>FETCH-LOGICAL-c471t-fdc37205164ece81709889add3ba302d06cbe881e72c849dd59e1cafb84f69c93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,3026,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10993810$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Knisley, Stephen B</creatorcontrib><creatorcontrib>Justice, Robert K</creatorcontrib><creatorcontrib>Kong, Wei</creatorcontrib><creatorcontrib>Johnson, Philip L</creatorcontrib><title>Ratiometry of transmembrane voltage-sensitive fluorescent dye emission in hearts</title><title>American journal of physiology. Heart and circulatory physiology</title><addtitle>Am J Physiol Heart Circ Physiol</addtitle><description>Department of Biomedical Engineering of the School of
Engineering, The University of Alabama at Birmingham, Alabama
35294-0019
Transmembrane voltage-sensitive
fluorescence measurements are limited by baseline drift that can
obscure changes in resting membrane potential and by motion artifacts
that can obscure repolarization. Voltage-dependent shift of emission
wavelengths may allow reduction of drift and motion artifacts by
emission ratiometry. We have tested this for action potentials and
potassium-induced changes in resting membrane potential in rabbit
hearts stained with di-4-ANEPPS [Pyridinium,
4-(2-(6-(dibutylamino)-2-naphthalenyl) ethenyl)-1-(3-sulfopropyl)-, hydroxide, inner salt] using laser excitation (488 nm) and a
two-photomultiplier tube system or spectrofluorometer (resolution of
500-1,000 Hz and <1 mm). Green and red emissions produced upright
and inverted action potentials, respectively. Ratios of green emission
to red emission followed action potential contours and exhibited larger fractional changes than either emission alone ( P < 0.001). The largest changes and signal-to-noise ratio (signal/noise)
were obtained with numerator wavelengths of 525-550 nm and
denominator wavelengths of 650-700 nm. Ratiometry lessened drift
56-66% ( P < 0.015) and indicated decreases in
resting membrane potential. Ratiometry lessened motion
artifacts and increased magnitudes of deflections representing
phase-zero depolarizations relative to total deflections by
123-188% in intact hearts ( P < 0.02). Durations
of action potentials at different pacing rates, temperatures, and
potassium concentrations were independent of whether they were measured
ratiometrically or with microelectrodes ( P 0.65). The
ratiometric calibration slope was 0.017/100 mV and decreased with time.
Thus emission ratiometry lessens the effects of motion and drift and
indicates resting membrane potential changes and repolarization.
potentiometric dye; Pyridinium,
4-(2-(6-(dibutylamino)-2-naphthalenyl) ethenyl)-1-(3-sulfopropyl)-,
hydroxide, inner salt; potassium; action potential</description><subject>Animals</subject><subject>Body Surface Potential Mapping</subject><subject>Cardiac Pacing, Artificial</subject><subject>Evaluation Studies as Topic</subject><subject>Fluorescent Dyes</subject><subject>In Vitro Techniques</subject><subject>Lasers</subject><subject>Membrane Potentials - drug effects</subject><subject>Membrane Potentials - physiology</subject><subject>Microelectrodes</subject><subject>Myocardium - metabolism</subject><subject>Potassium - metabolism</subject><subject>Potassium - pharmacology</subject><subject>Potentiometry - methods</subject><subject>Pyridinium Compounds</subject><subject>Rabbits</subject><subject>Spectrometry, Fluorescence - instrumentation</subject><subject>Spectrometry, Fluorescence - methods</subject><subject>Temperature</subject><issn>0363-6135</issn><issn>1522-1539</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kM1O3DAYRS1EBVPaV6jSDbtk_JM_qyuEClRCokKwthz7y8QoiVPboeTt8TDTaliw-ha-5_rqIPSd4IyQgq7l09SBdCGjGOOMVjxjWUdySo7QKr7TlBSMH6MVZiVLS8KKU_TZ-6cYLqqSnaBTgjlnNcEr9PteBmMHCG5JbJsEJ0c_wNDEC8mz7YPcQOph9CaYZ0jafrYOvIIxJHqBBAbjvbFjYsbkbZL_gj61svfwdX_P0OPVz4fLm_T27vrX5cVtqvKKhLTVilUUF6TMQUFNKszrmkutWSMZphqXqoG6JlBRVedc64IDUbJt6rwtueLsDJ3veidn_8zgg4hTFPR9HG5nLypKK8xyHIN8F1TOeu-gFZMzg3SLIFhsdYp_OsVWp4g6BRM3W52R_bb_ZG4G0Afkzl8MrHeBzmy6v8aBmLolCuntZjnofVf542Piau77B3gJ_9EDUky6Za-N2Jw9</recordid><startdate>20000901</startdate><enddate>20000901</enddate><creator>Knisley, Stephen B</creator><creator>Justice, Robert K</creator><creator>Kong, Wei</creator><creator>Johnson, Philip L</creator><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>7X8</scope></search><sort><creationdate>20000901</creationdate><title>Ratiometry of transmembrane voltage-sensitive fluorescent dye emission in hearts</title><author>Knisley, Stephen B ; Justice, Robert K ; Kong, Wei ; Johnson, Philip L</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c471t-fdc37205164ece81709889add3ba302d06cbe881e72c849dd59e1cafb84f69c93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Animals</topic><topic>Body Surface Potential Mapping</topic><topic>Cardiac Pacing, Artificial</topic><topic>Evaluation Studies as Topic</topic><topic>Fluorescent Dyes</topic><topic>In Vitro Techniques</topic><topic>Lasers</topic><topic>Membrane Potentials - drug effects</topic><topic>Membrane Potentials - physiology</topic><topic>Microelectrodes</topic><topic>Myocardium - metabolism</topic><topic>Potassium - metabolism</topic><topic>Potassium - pharmacology</topic><topic>Potentiometry - methods</topic><topic>Pyridinium Compounds</topic><topic>Rabbits</topic><topic>Spectrometry, Fluorescence - instrumentation</topic><topic>Spectrometry, Fluorescence - methods</topic><topic>Temperature</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Knisley, Stephen B</creatorcontrib><creatorcontrib>Justice, Robert K</creatorcontrib><creatorcontrib>Kong, Wei</creatorcontrib><creatorcontrib>Johnson, Philip L</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>American journal of physiology. Heart and circulatory physiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Knisley, Stephen B</au><au>Justice, Robert K</au><au>Kong, Wei</au><au>Johnson, Philip L</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Ratiometry of transmembrane voltage-sensitive fluorescent dye emission in hearts</atitle><jtitle>American journal of physiology. Heart and circulatory physiology</jtitle><addtitle>Am J Physiol Heart Circ Physiol</addtitle><date>2000-09-01</date><risdate>2000</risdate><volume>279</volume><issue>3</issue><spage>H1421</spage><epage>H1433</epage><pages>H1421-H1433</pages><issn>0363-6135</issn><eissn>1522-1539</eissn><abstract>Department of Biomedical Engineering of the School of
Engineering, The University of Alabama at Birmingham, Alabama
35294-0019
Transmembrane voltage-sensitive
fluorescence measurements are limited by baseline drift that can
obscure changes in resting membrane potential and by motion artifacts
that can obscure repolarization. Voltage-dependent shift of emission
wavelengths may allow reduction of drift and motion artifacts by
emission ratiometry. We have tested this for action potentials and
potassium-induced changes in resting membrane potential in rabbit
hearts stained with di-4-ANEPPS [Pyridinium,
4-(2-(6-(dibutylamino)-2-naphthalenyl) ethenyl)-1-(3-sulfopropyl)-, hydroxide, inner salt] using laser excitation (488 nm) and a
two-photomultiplier tube system or spectrofluorometer (resolution of
500-1,000 Hz and <1 mm). Green and red emissions produced upright
and inverted action potentials, respectively. Ratios of green emission
to red emission followed action potential contours and exhibited larger fractional changes than either emission alone ( P < 0.001). The largest changes and signal-to-noise ratio (signal/noise)
were obtained with numerator wavelengths of 525-550 nm and
denominator wavelengths of 650-700 nm. Ratiometry lessened drift
56-66% ( P < 0.015) and indicated decreases in
resting membrane potential. Ratiometry lessened motion
artifacts and increased magnitudes of deflections representing
phase-zero depolarizations relative to total deflections by
123-188% in intact hearts ( P < 0.02). Durations
of action potentials at different pacing rates, temperatures, and
potassium concentrations were independent of whether they were measured
ratiometrically or with microelectrodes ( P 0.65). The
ratiometric calibration slope was 0.017/100 mV and decreased with time.
Thus emission ratiometry lessens the effects of motion and drift and
indicates resting membrane potential changes and repolarization.
potentiometric dye; Pyridinium,
4-(2-(6-(dibutylamino)-2-naphthalenyl) ethenyl)-1-(3-sulfopropyl)-,
hydroxide, inner salt; potassium; action potential</abstract><cop>United States</cop><pmid>10993810</pmid><doi>10.1152/ajpheart.2000.279.3.h1421</doi></addata></record> |
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| ispartof | American journal of physiology. Heart and circulatory physiology, 2000-09, Vol.279 (3), p.H1421-H1433 |
| issn | 0363-6135 1522-1539 |
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| recordid | cdi_pubmed_primary_10993810 |
| source | MEDLINE; American Physiological Society; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals |
| subjects | Animals Body Surface Potential Mapping Cardiac Pacing, Artificial Evaluation Studies as Topic Fluorescent Dyes In Vitro Techniques Lasers Membrane Potentials - drug effects Membrane Potentials - physiology Microelectrodes Myocardium - metabolism Potassium - metabolism Potassium - pharmacology Potentiometry - methods Pyridinium Compounds Rabbits Spectrometry, Fluorescence - instrumentation Spectrometry, Fluorescence - methods Temperature |
| title | Ratiometry of transmembrane voltage-sensitive fluorescent dye emission in hearts |
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