Name: multiparametric optical mapping of the langendorff-perfused rabbit heart
Uploaded: 2011-09-13T12:00:00
Duration: 9 min 24 s
Description: Watch this Scientific Journal Video about Multiparametric Optical Mapping of the Langendorff-perfused Rabbit Heart at JoVE.com

एनआईएच HL085369 R01 अनुदान, HL067322, HL082729, EB008999

<ol>
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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Panoramic+imaging+reveals+basic+mechanisms+of+induction+and+termination+of+ventricular+tachycardia+in+rabbit+heart+with+chronic+infarction:+implications+for+low-voltage+cardioversion.">Panoramic imaging reveals basic mechanisms of induction and termination of ventricular tachycardia in rabbit heart with chronic infarction: implications for low-voltage cardioversion.</a> Heart Rhythm. 6, 87-97 (2009).</li><li>Efimov, I. R., Rendt, J. M., Salama, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+maps+of+intracellular+[Ca2+]i+transients+and+action-potentials+from+the+surface+of+perfused+guinea-pig+hearts+(abstract).">Optical maps of intracellular [Ca2+]i transients and action-potentials from the surface of perfused guinea-pig hearts (abstract).</a> Circulation. 90, 632-632 (1994).</li><li>Choi, B. R., Salama, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+maps+of+optical+action+potentials+and+calcium+transients+in+guinea-pig+hearts:+mechanisms+underlying+concordant+alternans.">Simultaneous maps of optical action potentials and calcium transients in guinea-pig hearts: mechanisms underlying concordant alternans.</a> J Physiol. 529, 171-188 (2000).</li><li>Fast, V. G., Ideker, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+optical+mapping+of+transmembrane+potential+and+intracellular+calcium+in+myocyte+cultures.">Simultaneous optical mapping of transmembrane potential and intracellular calcium in myocyte cultures.</a> J Cardiovasc Electrophysiol. 11, 547-556 (2000).</li><li>Laurita, K. R., Singal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+action+potentials+and+calcium+transients+simultaneously+from+the+intact+heart.">Mapping action potentials and calcium transients simultaneously from the intact heart.</a> Am J Physiol Heart Circ Physiol. 280, 2053-2060 (2001).</li><li>Choi, B. R., Burton, F., Salama, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytosolic+Ca2++triggers+early+afterdepolarizations+and+Torsade+de+Pointes+in+rabbit+hearts+with+type+2+long+QT+syndrome.">Cytosolic Ca2+ triggers early afterdepolarizations and Torsade de Pointes in rabbit hearts with type 2 long QT syndrome.</a> J Physiol. 543, 615-631 (2002).</li><li>Hwang, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+calcium+and+vulnerability+to+fibrillation+and+defibrillation+in+Langendorff-perfused+rabbit+ventricles.">Intracellular calcium and vulnerability to fibrillation and defibrillation in Langendorff-perfused rabbit ventricles.</a> Circulation. 114, 2595-2603 (2006).</li><li>Lou, Q., Efimov, I. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhanced+susceptibility+to+alternans+in+a+rabbit+model+of+chronic+myocardial+infarction.">Enhanced susceptibility to alternans in a rabbit model of chronic myocardial infarction.</a> Conf Proc IEEE Eng Med Biol Soc. , 4527-4530 (2009).</li><li>Kolega, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phototoxicity+and+photoinactivation+of+blebbistatin+in+UV+and+visible+light.">Phototoxicity and photoinactivation of blebbistatin in UV and visible light.</a> Biochem Biophys Res Commun. 320, 1020-1025 (2004).</li><li>Bray, M. A., Wikswo, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Considerations+in+phase+plane+analysis+for+nonstationary+reentrant+cardiac+behavior.">Considerations in phase plane analysis for nonstationary reentrant cardiac behavior.</a> Phys Rev E Stat Nonlin Soft Matter Phys. 65, 051902-05 (2002).</li><li>Fast, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recording+action+potentials+using+voltage-sensitive+dyes.">Recording action potentials using voltage-sensitive dyes.</a> Practical methods in cardiovascular research. , 233-255 (2005).</li><li>Knisley, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transmembrane+voltage+changes+during+unipolar+stimulation+of+rabbit+ventricle.">Transmembrane voltage changes during unipolar stimulation of rabbit ventricle.</a> Circ Res. 77, 1229-1239 (1995).</li><li>Bishop, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+photon+scattering+in+optical+signal+distortion+during+arrhythmia+and+defibrillation.">The role of photon scattering in optical signal distortion during arrhythmia and defibrillation.</a> Biophys J. 93, 3714-3726 (2007).</li><li>Efimov, I. R., Fedorov, V. V., Joung, B., Lin, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+cardiac+pacemaker+circuits:+methodological+puzzles+of+the+sinoatrial+node+optical+mapping.">Mapping cardiac pacemaker circuits: methodological puzzles of the sinoatrial node optical mapping.</a> Circ Res. 106, 255-271 (2010).</li><li>Fedorov, V. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+blebbistatin+as+an+excitation-contraction+uncoupler+for+electrophysiologic+study+of+rat+and+rabbit+hearts.">Application of blebbistatin as an excitation-contraction uncoupler for electrophysiologic study of rat and rabbit hearts.</a> Heart Rhythm. 4, 619-626 (2007).</li></ol>

ब्याज की कोई संघर्ष की घोषणा की.

हमारे अनुभव के आधार पर, एक सफल खरगोश Langendorff perfused दिल प्रयोग के लिए कुंजी समाधान अच्छी तरह से तैयार &#39;Tyrodes, दिल की जल्दी फसल अच्छी तरह से बनाए रखा छिड़काव दबाव, और छिड़काव प्रणाली में oxygenated समाधान के उचित पीएच शामिल हैं. आदेश में उच्चतम संभव संकेत से शोर अनुपात के साथ संकेत रिकॉर्ड करने के लिए, हम प्रकाश स्रोत, ऑप्टिकल फिल्टर, प्रकाशिकी ध्यान केंद्रित, photodetectors, 19 आदि सहित कारकों पर विचार करने की जरूरत है. इन पहलुओं का विवरण कहीं 19 से चर्चा कर रहे हैं . युवा खरगोश (उम्र: 4-5 महीने, वजन: 7-9 पाउंड) epicardial वसा, जो ऑप्टिकल संकेतों के शोर अनुपात करने के लिए संकेत कम हो जाती है से बचने के लिए इस्तेमाल किया जा सकता है. प्रत्येक पिक्सेल द्वारा दर्ज संकेत के ऊतक के एक मात्रा से उत्सर्जित प्रकाश की एक भारित एकीकरण है. इस ऊतक की मात्रा के गहराई इस्तेमाल डाई की उत्तेजना और उत्सर्जन तरंगदैर्य पर निर्भर करता है. Di चार - ANEPPS के लिए, एक उदाहरण के रूप में अनुमानित प्रवेश गहराई खरगोश 20 दिल में 300μm है. इस प्रकार, ऑप्टिकल संकेत की व्याख्या सावधानी के साथ किया जाना चाहिए जब बिजली समारोह के स्थानीय विविधता sinoatrial नोड, अलिंदनिलय संबंधी नोड में मौजूद हैं, और वेंट्रिकुलर अतालता 1,21,22 के दौरान . इलेक्ट्रोड रिकॉर्डिंग के साथ तुलना में ऑप्टिकल मानचित्रण तकनीक की एक सीमा है कि ऑप्टिकल संभावित कार्रवाई के repolarization चरण अक्सर हृदय संकुचन के कारण गति artifact के द्वारा distored है. यांत्रिक बाधा artifact के कम इस्तेमाल किया जा सकता है, लेकिन यह खत्म नहीं कर सकते हैं पूरी तरह. इसकी तुलना में, औषधीय उत्तेजना - संकुचन uncouplers गति artifact के हटाने में प्रभावी रहे हैं. हालांकि, इन uncouplers (जैसे Monoxime 2,3 Butanedione -) महत्वपूर्ण electrophysiological साइड इफेक्ट हो सकता है. Blebbistatin सामान्य 23 दिल में हृदय इलेक्ट्रोफिजियोलॉजी पर कोई प्रतिकूल दुष्प्रभाव प्रदर्शन किया गया था, और इस प्रकार ऑप्टिकल मानचित्रण के लिए एक आशाजनक uncoupler है. यह ध्यान दिया जाना चाहिए कि संकुचन के समाप्त करने के लिए कारण edema के त्वरण भी इलेक्ट्रोफिजियोलॉजी को प्रभावित कर सकता है.

<table border="1"><tbody><tr><td> अभिकर्मक </td><td> कंपनी </td><td> सूचीपत्र संख्या </td></tr><tr><td> NaCl </td><td> फिशर साइंटिफिक, फेयर लॉन, न्यू जर्सी </td><td> S271 - 1 </td></tr><tr><td> 2 CaCl (2H 2 हे) </td><td> फिशर साइंटिफिक, फेयर लॉन, न्यू जर्सी </td><td> C79-500 </td></tr><tr><td> KCl </td><td> फिशर साइंटिफिक, फेयर लॉन, न्यू जर्सी </td><td> S217 - 500 </td></tr><tr><td> 2 MgCl (6 2 हे) </td><td> फिशर साइंटिफिक, फेयर लॉन, न्यू जर्सी </td><td> M33-500 </td></tr><tr><td> नाह पीओ 4 (2 एच ओ) 2 </td><td> फिशर साइंटिफिक, फेयर लॉन, न्यू जर्सी </td><td> S369 - 500 </td></tr><tr><td> 3 NaHCO </td><td> फिशर साइंटिफिक, फेयर लॉन, न्यू जर्सी </td><td> S233 - 3 </td></tr><tr><td> डी ग्लुकोज </td><td> फिशर साइंटिफिक, फेयर लॉन, न्यू जर्सी </td><td> D16-1 </td></tr><tr><td> Blebbistatin </td><td> बायोसाइंस Tocris, Ellisville, MO </td><td> 1760 </td></tr><tr><td> डि चार - ANEPPS </td><td> Invitrogen, Carlsbad, CA </td><td> D1199 </td></tr><tr><td> RH237 </td><td> Invitrogen, Carlsbad, CA </td><td> S1109 </td></tr><tr><td> Rhod - 02:00 </td><td> Invitrogen, Carlsbad, CA </td><td> R1244 </td></tr><tr><td> Pluronic F127 </td><td> Invitrogen, Carlsbad, CA </td><td> P3000MP </td></tr><tr><td> डाइमिथाइल sulphoxide (DMSO) </td><td> सिग्मा, सेंट लुइस, MO </td><td> D2650 </td></tr></tbody></table>

multiparametric optical mapping of the langendorff-perfused rabbit heart

Optical imaging and fluorescent probes have significantly advanced research methodology in the field of cardiac electrophysiology in ways that could not have been accomplished by other approaches1. With the use of the calcium- and voltage-sensitive dyes, optical mapping allows measurement of transmembrane action potentials and calcium transients with high spatial resolution without the physical contact with the tissue. This makes measurements of the cardiac electrical activity possible under many conditions where the use of electrodes is inconvenient or impossible1. For example, optical recordings provide accurate morphological changes of membrane potential during and immediately after stimulation and defibrillation, while conventional electrode techniques suffer from stimulus-induced artifacts during and after stimuli due to electrode polarization1. 
The Langendorff-perfused rabbit heart is one of the most studied models of human heart physiology and pathophysiology. Many types of arrhythmias observed clinically could be recapitulated in the rabbit heart model. It was shown that wave patterns in the rabbit heart during ventricular arrhythmias, determined by effective size of the heart and the wavelength of reentry, are very similar to that in the human heart2. It was also shown that critical aspects of excitation-contraction (EC) coupling in rabbit myocardium, such as the relative contribution of sarcoplasmic reticulum (SR), is very similar to human EC coupling3. Here we present the basic procedures of optical mapping experiments in Langendorff-perfused rabbit hearts, including the Langendorff perfusion system setup, the optical mapping systems setup, the isolation and cannulation of the heart, perfusion and dye-staining of the heart, excitation-contraction uncoupling, and collection of optical signals. These methods could be also applied to the heart from species other than rabbit with adjustments to flow rates, optics, solutions, etc.
Two optical mapping systems are described. The panoramic mapping system is used to map the entire epicardium of the rabbit heart4-7. This system provides a global view of the evolution of reentrant circuits during arrhythmogenesis and defibrillation, and has been used to study the mechanisms of arrhythmias and antiarrhythmia therapy8,9. The dual mapping system is used to map the action potential (AP) and calcium transient (CaT) simultaneously from the same field of view10-13. This approach has enhanced our understanding of the important role of calcium in the electrical alternans and the induction of arrhythmia14-16.

Watch this Scientific Journal Video about Multiparametric Optical Mapping of the Langendorff-perfused Rabbit Heart at JoVE.com

Multiparametric Optical Mapping of the Langendorff-perfused Rabbit Heart

यह आलेख Langendorff - perfused खरगोश नयनाभिराम इमेजिंग प्रणाली का उपयोग कर दिल में ऑप्टिकल मानचित्रण प्रयोगों का आयोजन करने के लिए बुनियादी प्रक्रियाओं, और दोहरी वोल्टेज (और कैल्शियम) इमेजिंग साधन का वर्णन करता है.

Multiparametric खरगोश Langendorff perfused हार्ट के ऑप्टिकल मानचित्रण

Optical imaging and fluorescent probes have significantly advanced research methodology in the field of cardiac electrophysiology in ways that could not ...

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