| A novel approach to dual excitation ratiometric optical mapping of cardiac action potentials with di-4-ANEPPS using pulsed LED excitation. | |
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MedLine Citation:
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PMID: 21536528 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We developed a new method for ratiometric optical mapping of transmembrane potential (V(m)) in cardiac preparations stained with di-4-ANEPPS. V(m)-dependent shifts of excitation and emission spectra establish two excitation bands (<481 and >481 nm) that produce fluorescence changes of opposite polarity within a single emission band (575-620 nm). The ratio of these positive and negative fluorescence signals (excitation ratiometry) increases V(m) sensitivity and removes artifacts common to both signals. We pulsed blue (450 ± 10 nm) and cyan (505 ± 15 nm) light emitting diodes (LEDs) at 375 Hz in alternating phase synchronized to a camera (750 frames-per-second). Fluorescence was bandpass filtered (585 ± 20 nm). This produced signals with upright (blue) and inverted (cyan) action potentials (APs) interleaved in sequential frames. In four whole swine hearts with motion chemically arrested, fractional fluorescence for blue, cyan, and ratio signals was 1.2 ± 0.3%, 1.2 ± 0.3%, and 2.4 ± 0.6%, respectively. Signal-to-noise ratios were 4.3 ± 1.4, 4.0 ± 1.2, and 5.8 ± 1.9, respectively. After washing out the electromechanical uncoupling agent, we characterized motion artifact by cross-correlating blue, cyan, and ratio signals with a signal with normal AP morphology. Ratiometry improved cross-correlation coefficients from 0.50 ± 0.48 to 0.81 ± 0.25, but did not cancel all motion artifacts. These findings demonstrate the feasibility of pulsed LED excitation ratiometry in myocardium. |
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Authors:
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Andrew D Bachtel; Richard A Gray; Jayna M Stohlman; Elliot B Bourgeois; Andrew E Pollard; Jack M Rogers |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S. Date: 2011-05-02 |
Journal Detail:
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Title: IEEE transactions on bio-medical engineering Volume: 58 ISSN: 1558-2531 ISO Abbreviation: IEEE Trans Biomed Eng Publication Date: 2011 Jul |
Date Detail:
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Created Date: 2011-06-22 Completed Date: 2011-10-20 Revised Date: 2012-03-13 |
Medline Journal Info:
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Nlm Unique ID: 0012737 Medline TA: IEEE Trans Biomed Eng Country: United States |
Other Details:
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Languages: eng Pagination: 2120-6 Citation Subset: IM |
Copyright Information:
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© 2011 IEEE |
Affiliation:
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Covidien, Boulder, CO 80301, USA. andrew.bachtel@covidien.com |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Action Potentials
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physiology* Animals Artifacts Female Heart / physiology* Male Pyridinium Compounds / chemistry* Signal Processing, Computer-Assisted Subtraction Technique Swine Voltage-Sensitive Dye Imaging / methods* |
| Grant Support | |
ID/Acronym/Agency:
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HL64184/HL/NHLBI NIH HHS; R01 HL092049-04/HL/NHLBI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Pyridinium Compounds; 90134-00-2/1-(3-sulfonatopropyl)-4-(beta)(2-(di-n-butylamino)-6-naphthylvinyl)pyridinium betaine |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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