Name: local field fluorescence microscopy: imaging cellular signals in intact hearts
Uploaded: 2017-03-08T13:00:00
Description: Watch this Scientific Journal Video about Local Field Fluorescence Microscopy: Imaging Cellular Signals in Intact Hearts at JoVE.com

Мы благодарим доктора Алисии Mattiazzi для критического обсуждения представленной работы. Эта работа была поддержана грантом от NIH (R01 HL-084487) для ALE.

Этот протокол и все обработки мышей был одобрен Institutional уходу и использованию животных комитета UC Мерсед (№ 2008-201) путем. Были проведены эксперименты с попугаи в 1999 году в соответствии с общей политикой для использования животных, установленной научной комиссией венесуэльского институ...

<ol>
	<li>Fabiato, A., Fabiato, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contractions+induced+by+a+calcium-triggered+release+of+calcium+from+the+sarcoplasmic+reticulum+of+single+skinned+cardiac+cells.">Contractions induced by a calcium-triggered release of calcium from the sarcoplasmic reticulum of single skinned cardiac cells.</a> J Physiol. 249 (3), 469-495 (1975).</li><li>Mitra, R., Morad, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+uniform+enzymatic+method+for+dissociation+of+myocytes+from+hearts+and+stomachs+of+vertebrates.">A uniform enzymatic method for dissociation of myocytes from hearts and stomachs of vertebrates.</a> Am J Physiol. 249 (5), 1056-1060 (1985).</li><li>Escobar, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+changes+of+intracellular+Ca2++transients+in+beating+rat+hearts.">Developmental changes of intracellular Ca2+ transients in beating rat hearts.</a> Am J Physiol Heart Circ Physiol. 286 (3), H971-H978 (2004).</li><li>Ramos-Franco, J., Aguilar-Sanchez, Y., Escobar, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intact+Heart+Loose+Patch+Photolysis+Reveals+Ionic+Current+Kinetics+During+Ventricular+Action+Potentials.">Intact Heart Loose Patch Photolysis Reveals Ionic Current Kinetics During Ventricular Action Potentials.</a> Circ Res. 118 (2), 203-215 (2016).</li><li>Mitra, R., Morad, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+uniform+enzymatic+method+for+dissociation+of+myocytes+from+hearts+and+stomachs+of+vertebrates.">A uniform enzymatic method for dissociation of myocytes from hearts and stomachs of vertebrates.</a> Am J Physiol. 249, H1056-H1060 (1985).</li><li>Fischmeister, R., DeFelice, L. J., Ayer, R. K., Levi, R., DeHaan, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Channel+currents+during+spontaneous+action+potentials+in+embryonic+chick+heart+cells.+The+action+potential+patch+clamp.">Channel currents during spontaneous action potentials in embryonic chick heart cells. The action potential patch clamp.</a> Biophys J. 46 (2), 267-271 (1984).</li><li>Banyasz, T., Horvath, B., Jian, Z., Izu, L. T., Chen-Izu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Profile+of+L-type+Ca(2+)+current+and+Na(+)/Ca(2+)+exchange+current+during+cardiac+action+potential+in+ventricular+myocytes.">Profile of L-type Ca(2+) current and Na(+)/Ca(2+) exchange current during cardiac action potential in ventricular myocytes.</a> Heart Rhythm. 9 (1), 134-142 (2012).</li><li>Apkon, M., Nerbonne, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+two+distinct+depolarization-activated+K++currents+in+isolated+adult+rat+ventricular+myocytes.">Characterization of two distinct depolarization-activated K+ currents in isolated adult rat ventricular myocytes.</a> J Gen Physiol. 97 (5), 973-1011 (1991).</li><li>Valverde, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transient+Ca2++depletion+of+the+sarcoplasmic+reticulum+at+the+onset+of+reperfusion.">Transient Ca2+ depletion of the sarcoplasmic reticulum at the onset of reperfusion.</a> Cardiovasc Res. 85 (4), 671-680 (2010).</li><li>Valverde, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phospholamban+phosphorylation+sites+enhance+the+recovery+of+intracellular+Ca2++after+perfusion+arrest+in+isolated,+perfused+mouse+heart.">Phospholamban phosphorylation sites enhance the recovery of intracellular Ca2+ after perfusion arrest in isolated, perfused mouse heart.</a> Cardiovasc Res. 70 (2), 335-345 (2006).</li><li>Escobar, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+inositol+1,4,5-trisphosphate+in+the+regulation+of+ventricular+Ca(2+)+signaling+in+intact+mouse+heart.">Role of inositol 1,4,5-trisphosphate in the regulation of ventricular Ca(2+) signaling in intact mouse heart.</a> J Mol Cell Cardiol. 53 (6), 768-779 (2012).</li><li>Mej&#237;a-Alvarez, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulsed+local-field+fluorescence+microscopy:+a+new+approach+for+measuring+cellular+signals+in+the+beating+heart.">Pulsed local-field fluorescence microscopy: a new approach for measuring cellular signals in the beating heart.</a> Pfl&#252;gers Arch. 445 (6), 747-758 (2003).</li><li>Ferreiro, M., Petrosky, A. D., Escobar, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracellular+Ca2++release+underlies+the+development+of+phase+2+in+mouse+ventricular+action+potentials.">Intracellular Ca2+ release underlies the development of phase 2 in mouse ventricular action potentials.</a> Am J Physiol Heart Circ Physiol. 302 (5), H1160-H1172 (2012).</li><li>Kornyeyev, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calsequestrin+2+deletion+shortens+the+refractoriness+of+Ca2++release+and+reduces+rate-dependent+Ca2+-alternans+in+intact+mouse+hearts.">Calsequestrin 2 deletion shortens the refractoriness of Ca2+ release and reduces rate-dependent Ca2+-alternans in intact mouse hearts.</a> J Mol Cell Cardiol. 52 (1), 21-31 (2012).</li><li>Mattiazzi, A., Argenziano, M., Aguilar-Sanchez, Y., Mazzocchi, G., Escobar, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ca2++Sparks+and+Ca2++waves+are+the+subcellular+events+underlying+Ca2++overload+during+ischemia+and+reperfusion+in+perfused+intact+hearts.">Ca2+ Sparks and Ca2+ waves are the subcellular events underlying Ca2+ overload during ischemia and reperfusion in perfused intact hearts.</a> J Mol Cell Cardiol. 79, 69-78 (2015).</li><li>Kornyeyev, D., Reyes, M., Escobar, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Luminal+Ca(2+)+content+regulates+intracellular+Ca(2+)+release+in+subepicardial+myocytes+of+intact+beating+mouse+hearts:+effect+of+exogenous+buffers.">Luminal Ca(2+) content regulates intracellular Ca(2+) release in subepicardial myocytes of intact beating mouse hearts: effect of exogenous buffers.</a> Am J Physiol Heart Circ Physiol. 298 (6), H2138-H2153 (2010).</li><li>Wang, Z. G., Fermini, B., Nattel, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repolarization+differences+between+guinea+pig+atrial+endocardium+and+epicardium:+evidence+for+a+role+of+Ito.">Repolarization differences between guinea pig atrial endocardium and epicardium: evidence for a role of Ito.</a> Am J Physiol. 260, H1501-H1506 (1991).</li><li>Liu, D. W., Gintant, G. A., Antzelevitch, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ionic+bases+for+electrophysiological+distinctions+among+epicardial,+midmyocardial,+and+endocardial+myocytes+from+the+free+wall+of+the+canine+left+ventricle.">Ionic bases for electrophysiological distinctions among epicardial, midmyocardial, and endocardial myocytes from the free wall of the canine left ventricle.</a> Circ Res. 72 (3), 671-687 (1993).</li><li>Litovsky, S. H., Antzelevitch, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transient+outward+current+prominent+in+canine+ventricular+epicardium+but+not+endocardium.">Transient outward current prominent in canine ventricular epicardium but not endocardium.</a> Circ Res. 62 (1), 116-126 (1988).</li><li>Lukas, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiology+of+Myocardial+Cells+in+the+Epicardial,+Midmyocardial,+and+Endocardial+Layers+of+the+Ventricle.">Electrophysiology of Myocardial Cells in the Epicardial, Midmyocardial, and Endocardial Layers of the Ventricle.</a> J Cardiovasc Pharmacol Ther. 2 (1), 61-72 (1997).</li><li>Antzelevitch, C., Fish, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+heterogeneity+within+the+ventricular+wall.">Electrical heterogeneity within the ventricular wall.</a> Basic Res Cardiol. 96 (6), 517-527 (2001).</li><li>Gomez, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+electrical+heterogeneity+by+compensated+hypertrophy+in+rat+left+ventricle.">Modulation of electrical heterogeneity by compensated hypertrophy in rat left ventricle.</a> Am J Physiol. 272 (3 Pt 2), H1078-H1086 (1997).</li><li>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=Innovation+in+optical+imaging:+looking+inside+the+heart.">Innovation in optical imaging: looking inside the heart.</a> Heart Rhythm. 4 (7), 925-926 (2007).</li><li>Boukens, B. J., 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=A+century+of+optocardiography.">A century of optocardiography.</a> IEEE Rev Biomed Eng. 7, 115-125 (2014).</li><li>Zhang, H., Iijima, K., Huang, J., Walcott, G. P., Rogers, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+mapping+of+membrane+potential+and+epicardial+deformation+in+beating+hearts.">Optical mapping of membrane potential and epicardial deformation in beating hearts.</a> Biophysics J. 111 (2), 438-451 (2016).</li></ol>

Эта статья сосредоточена в описании локальных методов поля флуоресценции оценить функцию кардиомиоцитов бывших естественных условиях. Изучение этих клеток в сочетании среде не только более физиологические, но также очень подходит для оценки патологии органа уровня. Клеточные с...

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Sodium chloride</td>
			<td>Sigma</td>
			<td>7647-14-5</td>
			<td></td>
		</tr>
		<tr>
			<td>D-(+)-glucose</td>
			<td>Sigma</td>
			<td>50-99-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium chloride</td>
			<td>Sigma</td>
			<td>7447-40-7</td>
			<td></td>
		</tr>
		<tr>
			<td>HEPES</td>
			<td>Sigma</td>
			<td>7365-45-9</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium phosphate</td>
			<td>Sigma</td>
			<td>10049-21-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium chloride solution</td>
			<td>Sigma</td>
			<td>10043-52-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium chloride solution</td>
			<td>Sigma</td>
			<td>7786-30-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium hyrdoxide</td>
			<td>Sigma</td>
			<td>S-8045</td>
			<td></td>
		</tr>
		<tr>
			<td>0.2um nylon membrane filter</td>
			<td>Whatman</td>
			<td>7402-004</td>
			<td></td>
		</tr>
		<tr>
			<td>Manifold MPP</td>
			<td>Warner</td>
			<td>64-0216&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>21G1.5 Precision glide needle</td>
			<td>B-D</td>
			<td>305167</td>
			<td></td>
		</tr>
		<tr>
			<td>Black Braided silk string (non-absorbable surgical suture)</td>
			<td>AllMech Tech</td>
			<td>LOOK-SP105</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin sodium injection 1000USP/mL</td>
			<td>AllMech Tech</td>
			<td>NDC63323-540-11</td>
			<td></td>
		</tr>
		<tr>
			<td>DMSO D8779</td>
			<td>Sigma</td>
			<td>67-68-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Blebbistatin</td>
			<td>Sigma</td>
			<td>856925-71-8</td>
			<td></td>
		</tr>
		<tr>
			<td>Pluronic F-127 20% solution</td>
			<td>Biotium</td>
			<td>59004</td>
			<td></td>
		</tr>
		<tr>
			<td>Materflex C/L peristaltic pump</td>
			<td>Cole-Parmer</td>
			<td>77122-26</td>
			<td></td>
		</tr>
		<tr>
			<td>Isostim stimulator</td>
			<td>World Precision Instruments</td>
			<td>A320RC</td>
			<td></td>
		</tr>
		<tr>
			<td>Waveform generator</td>
			<td>Teledyne Lecroy</td>
			<td>WaveStation 2012</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasonic cleaner FS20</td>
			<td>Fisher Scientific</td>
			<td>1533530</td>
			<td></td>
		</tr>
		<tr>
			<td>Tygon tubing ID:1/32&#34; OD:3/32&#34; Wall 1/32&#34;</td>
			<td>Component Supply</td>
			<td>TET-031A</td>
			<td></td>
		</tr>
		<tr>
			<td>Tygon tubing ID:3/32&#34; OD:5/32&#34; Wall 1/32&#34;</td>
			<td>Component Supply</td>
			<td>TET-094A</td>
			<td></td>
		</tr>
		<tr>
			<td>Adapter luer lock to 3-way valve</td>
			<td>Cole-Parmer</td>
			<td>EW-31200-80</td>
			<td></td>
		</tr>
		<tr>
			<td>Tee adapters and plastic fittings</td>
			<td>Cole-Parmer</td>
			<td>6365-90</td>
			<td></td>
		</tr>
		<tr>
			<td>Plastic clamp</td>
			<td>WaterZoo</td>
			<td>2465</td>
			<td></td>
		</tr>
		<tr>
			<td>Peltier</td>
			<td>TE Technology</td>
			<td>TE-127-2.0-2.5</td>
			<td></td>
		</tr>
		<tr>
			<td>Rhod-2AM</td>
			<td>ThermoFisher Scientific</td>
			<td>R1245MP</td>
			<td></td>
		</tr>
		<tr>
			<td>Di-8-ANEPPS</td>
			<td>ThermoFisher Scientific</td>
			<td>D3167</td>
			<td></td>
		</tr>
		<tr>
			<td>Mag-Fluo-4AM</td>
			<td>ThermoFisher Scientific</td>
			<td>M14206</td>
			<td></td>
		</tr>
		<tr>
			<td>Acupunture needles</td>
			<td>LHASA</td>
			<td>TC1.20x13</td>
			<td></td>
		</tr>
		<tr>
			<td>60mL BD syringe with luer-lok</td>
			<td>Fisher Scientific</td>
			<td>14-820-11</td>
			<td></td>
		</tr>
		<tr>
			<td>LabView</td>
			<td>National Instruments</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Speed vacuum Eppendorf Vagufuge</td>
			<td>Fisher Scientific</td>
			<td>07-748-13</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital signal processing circuit DSP TMS 320</td>
			<td>Texas Instrument</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Longpass Dichroic mirror 567nm</td>
			<td>ThorLabs</td>
			<td>DMLP567L</td>
			<td></td>
		</tr>
		<tr>
			<td>Objective 10X NA 0.25 DIN AchromaticFinite Intl Standard Objective</td>
			<td>Edmund Optics</td>
			<td>Stock# 33-437</td>
			<td></td>
		</tr>
		<tr>
			<td>Objective 20X NA 0.40 DIN Achromatic Finite Intl Standard Objective</td>
			<td>Edmund Optics</td>
			<td>Stock# 33-438</td>
			<td></td>
		</tr>
		<tr>
			<td>Longpass colored glass filter 590 nm&#160;</td>
			<td>ThorLabs</td>
			<td>FGL590</td>
			<td></td>
		</tr>
		<tr>
			<td>Green Nd-YAG laser 532nm, 500mW</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micromanipulator for laser</td>
			<td>Siskiyou</td>
			<td>MX130R</td>
			<td></td>
		</tr>
		<tr>
			<td>Multimode fiber optic 200&#181;m NA 0.39</td>
			<td>ThorLabs</td>
			<td>FT200UMT</td>
			<td></td>
		</tr>
		<tr>
			<td>LEDs blue</td>
			<td>Lumileds</td>
			<td colspan="2">L135-B475003500000</td>
		</tr>
		<tr>
			<td>LEDs green</td>
			<td>Lumileds</td>
			<td colspan="2">L135-G525003500000</td>
		</tr>
		<tr>
			<td>Cube beam splitter, non-polarizing</td>
			<td>ThorLabs</td>
			<td>BS007</td>
			<td></td>
		</tr>
		<tr>
			<td>36&#34; Length, Dovetail Optical Rail</td>
			<td>Edmund Optics</td>
			<td>54-402</td>
			<td></td>
		</tr>
		<tr>
			<td>2.5&#34; Width, Dovetail Carrier</td>
			<td>Edmund Optics</td>
			<td>54-404</td>
			<td></td>
		</tr>
		<tr>
			<td>0.75&#34; Travel, micrometer stage</td>
			<td>Edmund Optics</td>
			<td>37-983</td>
			<td></td>
		</tr>
		<tr>
			<td>Dovetail optical rail 3&#34;</td>
			<td>ThorLabs</td>
			<td>RLA075/M</td>
			<td></td>
		</tr>
		<tr>
			<td>Dovetail rail carrier 1&#34;</td>
			<td>ThorLabs</td>
			<td>RC1</td>
			<td></td>
		</tr>
		<tr>
			<td>Avalanche photodiode Helix 902</td>
			<td>Digi-Key</td>
			<td>HELIX-902-200</td>
			<td></td>
		</tr>
		<tr>
			<td>Objective holder XY Translator&#160;</td>
			<td>ThorLabs</td>
			<td>ST1XY-S</td>
			<td></td>
		</tr>
		<tr>
			<td>Aluminum breadboard 6&#34;x6&#34;</td>
			<td>ThorLabs</td>
			<td>MB6</td>
			<td></td>
		</tr>
		<tr>
			<td>Nexus otpical table 4&#39;x6&#39;</td>
			<td>ThorLabs</td>
			<td>T46HK</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel cap screws</td>
			<td>ThorLabs</td>
			<td>HW-KIT5/M</td>
			<td></td>
		</tr>
		<tr>
			<td>Acrylic&#160; sheet</td>
			<td>Home Depot/Lowes</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sylgard Silicone elastomer kit</td>
			<td>Dow Corning</td>
			<td>Sylgard 184</td>
			<td></td>
		</tr>
		<tr>
			<td>Epoxy gel&#160;</td>
			<td>Walmart/Home Depot</td>
			<td>2-part 5 min clr 1oz</td>
			<td></td>
		</tr>
		<tr>
			<td>21G1.5 Precision glide needle</td>
			<td>B-D</td>
			<td>305167</td>
			<td></td>
		</tr>
		<tr>
			<td>23G Precision glide needle</td>
			<td>B-D</td>
			<td>305145</td>
			<td></td>
		</tr>
		<tr>
			<td>Mounting base, 25mmx75mmx10mm</td>
			<td>ThorLabs</td>
			<td>BA1/M</td>
			<td></td>
		</tr>
		<tr>
			<td>Wire shelve posts 36&#34;</td>
			<td>Alera</td>
			<td>AALESW59PO36SR</td>
			<td></td>
		</tr>
		<tr>
			<td>Wire shelves</td>
			<td>Alera</td>
			<td>ALESW582424SR</td>
			<td></td>
		</tr>
		<tr>
			<td>Post and angle clamp</td>
			<td>ThorLabs</td>
			<td>SWC/M-P5</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass syringe for dye chamber</td>
			<td>Wheaton</td>
			<td>W851020</td>
			<td></td>
		</tr>
		<tr>
			<td>Rubber stopper</td>
			<td>Home Science Tools</td>
			<td>CE-STOP01C</td>
			<td></td>
		</tr>
	</tbody>
</table>

local field fluorescence microscopy: imaging cellular signals in intact hearts

В сердце, исследования молекулярной сигнализации, как правило, выполняются в изолированных миоцитов. Тем не менее, многие патологические состояния, такие как ишемия и аритмий могут быть полностью поняты только на уровне целого органа. Здесь мы представляем метод спектроскопический локального поля флуоресцентной микроскопии (LFFM), что позволяет измерять клеточных сигналов в интактном сердце. Методика основана на комбинации Лангендорфа перфузию сердца и оптических волокон для записи флуоресцентные сигналы. LFFM имеет различные приложения в области сердечно-сосудистой физиологии для изучения сердца при нормальных и патологических состояниях. Несколько переменных сердца можно контролировать с помощью различных флуоресцентных индикаторов. К ним относятся цитозольного [Ca 2+], внутрисуставной саркоплазматического ретикулума [Ca 2+] и мембранных потенциалов. Экзогенные флуоресцентные зонды возбуждаются и излучаемый флуоресценции обнаружена с тремя различными расположений техники LFFM эпифлуоресцентнойы представлены в этой статье. Центральные различия между этими методами являются тип источника света, используемого для возбуждения и на пути света возбуждения модулируется. Импульсный LFFM (PLFFM) использует лазерные световые импульсы во время непрерывной волны LFFM (CLFFM) использует непрерывный лазерный луч для возбуждения. Наконец, светоизлучающие диоды (СИД) были использованы в качестве третьего источника света. Это некогерентного расположение называется импульсно светодиодной флуоресцентной микроскопии (PLEDFM).

AP и Са 2+ в эндокарда и эпикарда Для сравнения сигналов через стенку желудочка, один волоконно-оптический позиционируется в эндокарда, а другой в эпикарда. Сравнивая морфологию AP, записанной от эндокарда...

Watch this Scientific Journal Video about Local Field Fluorescence Microscopy: Imaging Cellular Signals in Intact Hearts at JoVE.com

Local Field Fluorescence Microscopy: Imaging Cellular Signals in Intact Hearts

В самом центре, молекулярные события координировать электрическую и сократительную функцию органа. Набор локальных методов микроскопии поля флуоресценции, представленные здесь позволяет записывать клеточных переменных в интактных сердцах. Выявление механизмов, определяющих функцию сердца имеет решающее значение для понимания того, как сердце работает в патологических ситуациях.

Локальное поле флуоресцентной микроскопией: Визуализация клеточных сигналов в интактных сердцах

In the heart, molecular signaling studies are usually performed in isolated myocytes. However, many pathological situations such as ischemia and ...

Research

JoVE Journal

Biology

Long-term Imaging Mammalian Cells using Wide-Field Microscopy

Долгосрочные изображений клетках млекопитающих использованием широкого поля микроскопии

Imaging Exocytosis in Retinal Bipolar Cells with TIRF Microscopy

Изображений экзоцитоза в клетки сетчатки Биполярный с Микроскопия TIRF

Total internal reflectance fluorescence (TIRF) microscopy is a technique that allows the study of events happening at the cell membrane, by selective ...

Time-lapse Imaging of Mitosis After siRNA Transfection

Покадровый Визуализация Митоз После трансфекции миРНК

Changes in cellular organization and chromosome dynamics that occur during mitosis are tightly coordinated to ensure accurate inheritance of genomic and ...

In vivo Imaging of Intact Drosophila Larvae at Sub-cellular Resolution

In vivo Imaging of Intact Drosophila Larvae at Sub-cellular Resolution

В естественных изображений интактного личинки дрозофилы на субклеточном Резолюцию

Recent improvements in optical imaging, genetically encoded fluorophores and genetic tools allowing efficient establishment of desired transgenic animal ...

Fluorescence-based Measurement of Store-operated Calcium Entry in Live Cells: from Cultured Cancer Cell to Skeletal Muscle Fiber

Флуоресценции на основе измерения магазин управлением поступления кальция в живых клетках: от искусственного раковых клеток в скелетных мышечных волокон

Store operated Ca2+ entry (SOCE), earlier termed capacitative Ca2+ entry, is a tightly regulated mechanism for influx of extracellular Ca2+ into cells to ...

Rapid Analysis and Exploration of Fluorescence Microscopy Images

Экспресс-анализ и разведка флуоресцентной микроскопии Images

Despite rapid advances in high-throughput microscopy, quantitative image-based assays still pose significant challenges. While a variety of specialized ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

В Vivo 4-мерном Tracking гемопоэтических стволовых и клеток-предшественников в взрослой мыши свода черепа костного мозга

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

Тандем Замораживание высокого давления и резких похолоданий Замена тканей растений для просвечивающей электронной микроскопии

Since the 1940s transmission electron microscopy (TEM) has been providing biologists with ultra-high resolution images of biological materials. Yet, ...

Imaging Local Ca2+ Signals in Cultured Mammalian Cells

Imaging Local Ca2+ Signals in Cultured Mammalian Cells

Изображений Локальный ЦС<sup&gt; 2+</sup&gt; Сигналы в культуре клетках млекопитающих

Cytosolic Ca2+ ions regulate numerous aspects of cellular activity in almost all cell types, controlling processes as wide-ranging as gene transcription, ...

Cellular Redox Profiling Using High-content Microscopy

Сотовая редокс-профилирование с использованием высококонцентрированной микроскопии

Reactive oxygen species (ROS) regulate essential cellular processes including gene expression, migration, differentiation and proliferation. However, ...