Name: time-resolved in vivo measurement of neuropeptide dynamics by capacitive immunoprobe in porcine heart
Uploaded: 2022-05-19T15:00:00
Description: Watch this Scientific Journal Video about Time-Resolved In Vivo Measurement of Neuropeptide Dynamics by Capacitive Immunoprobe in Porcine Heart at JoVE.com

我们感谢 Olu Ajijola 博士（加州大学洛杉矶分校心律失常中心）对 体内 实验的专家支持。这项工作得到了NIH U01 EB025138（JLA，CS）的支持。

所有动物实验均获得加州大学洛杉矶分校动物研究委员会的批准，并按照美国国立卫生研究院实验动物护理和使用指南（第8版，2011年）制定的指南进行。约75公斤的成年雄性约克郡猪用于 体内 研究10。
1. 电容式免疫探针制造和功能化
<ol><li>使用手术刀从一端切割25厘米长的全氟烷氧（PFA）涂层铂丝（见 材料表），并从一端...

<ol>
	<li>Chow, S. 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+Biomarkers+for+the+prevention,+assessment,+and+management+of+heart+failure:+A+scientific+statement+from+the+American+Heart+Association.">Role of Biomarkers for the prevention, assessment, and management of heart failure: A scientific statement from the American Heart Association.</a> Circulation. 135 (22), 1054-1091 (2017).</li><li>Goldstein, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adrenal+responses+to+stress.">Adrenal responses to stress.</a> Cellular and Molecular Neurobiology. 30 (8), 1433-1440 (2010).</li><li>Ullman, B., Hulting, J., Lundberg, 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=Prognostic+value+of+plasma+neuropeptide-Y+in+coronary+care+unit+patients+with+and+without+acute+myocardial+infarction.">Prognostic value of plasma neuropeptide-Y in coronary care unit patients with and without acute myocardial infarction.</a> European Heart Journal. 15 (4), 454-461 (1994).</li><li>Farrell, D. M., 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=Angiotensin+II+modulates+catecholamine+release+into+interstitial+fluid+of+canine+myocardium+in+vivo.">Angiotensin II modulates catecholamine release into interstitial fluid of canine myocardium in vivo.</a> American Journal of Physiology-Heart and Circulatory Physiology. 281 (2), 813-822 (2001).</li><li>Ardell, J. L., Foreman, R. D., Armour, J. A., Shivkumar, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+sympathectomy+and+spinal+cord+stimulation+attenuate+reflex-mediated+norepinephrine+release+during+ischemia+preventing+ventricular+fibrillation.">Cardiac sympathectomy and spinal cord stimulation attenuate reflex-mediated norepinephrine release during ischemia preventing ventricular fibrillation.</a> JCI Insight. 4 (23), 131648 (2019).</li><li>Franco-Cereceda, A., Lundberg, J. M., Dahlof, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuropeptide+Y+and+sympathetic+control+of+heart+contractility+and+coronary+vascular+tone.">Neuropeptide Y and sympathetic control of heart contractility and coronary vascular tone.</a> Acta Physiologica Scandinavica. 124 (3), 361-369 (1985).</li><li>Habecker, B. 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=Molecular+and+cellular+neurocardiology:+development,+and+cellular+and+molecular+adaptations+to+heart+disease.">Molecular and cellular neurocardiology: development, and cellular and molecular adaptations to heart disease.</a> The Journal of Physiology. 594 (14), 3853-3875 (2016).</li><li>Hoang, J. D., Salavatian, S., Yamaguchi, N., Swid, M. A., Vaseghi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+sympathetic+activation+circumvents+high-dose+beta+blocker+therapy+in+part+through+release+of+neuropeptide+Y.">Cardiac sympathetic activation circumvents high-dose beta blocker therapy in part through release of neuropeptide Y.</a> JCI Insight. 5 (11), 135519 (2020).</li><li>Rigel, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+neuropeptides+on+heart+rate+in+dogs:+comparison+of+VIP,+PHI,+NPY,+CGRP,+and+NT.">Effects of neuropeptides on heart rate in dogs: comparison of VIP, PHI, NPY, CGRP, and NT.</a> American Journal of Physiology. 255, 311-317 (1988).</li><li>Kluge, N., 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=Rapid+measurement+of+cardiac+neuropeptide+dynamics+by+capacitive+immunoprobe+in+the+porcine+heart.">Rapid measurement of cardiac neuropeptide dynamics by capacitive immunoprobe in the porcine heart.</a> American Journal of Physiology-Heart and Circulatory Physiology. 320 (1), 66-76 (2021).</li><li>Berggren, C., Bjarnason, B., Johansson, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capacitive+biosensors.">Capacitive biosensors.</a> Electroanalysis. 13 (3), 173-180 (2001).</li><li>Prodromidis, M. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impedimetric+immunosensors-A+review.">Impedimetric immunosensors-A review.</a> Electrochimica Acta. 55 (14), 4227-4233 (2010).</li><li>Lee, H., Dellatore, S. M., Miller, W. M., Messersmith, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mussel-inspired+surface+chemistry+for+multifunctional+coatings.">Mussel-inspired surface chemistry for multifunctional coatings.</a> Science. 318 (5849), 426-430 (2007).</li><li>Leszczyszyn, D. J., 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=Secretion+of+catecholamines+from+individual+adrenal+medullary+chromaffin+cells.">Secretion of catecholamines from individual adrenal medullary chromaffin cells.</a> Journal of Neurochemistry. 56 (6), 1855-1863 (1991).</li><li>Pihel, K., Schroeder, T. J., Wightman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+and+selective+cyclic+voltammetric+measurements+of+epinephrine+and+norepinephrine+as+a+method+to+measure+secretion+from+single+bovine+adrenal+medullary+cells.">Rapid and selective cyclic voltammetric measurements of epinephrine and norepinephrine as a method to measure secretion from single bovine adrenal medullary cells.</a> Analytical Chemistry. 66 (24), 4532-4537 (1994).</li><li>Jaffe, E. H., Marty, A., Schulte, A., Chow, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extrasynaptic+vesicular+transmitter+release+from+the+somata+of+substantia+nigra+neurons+in+rat+midbrain+slices.">Extrasynaptic vesicular transmitter release from the somata of substantia nigra neurons in rat midbrain slices.</a> The Journal of Neuroscience. 18 (10), 3548-3553 (1998).</li><li>Walsh, P. L., Petrovic, J., Wightman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinguishing+splanchnic+nerve+and+chromaffin+cell+stimulation+in+mouse+adrenal+slices+with+fast-scan+cyclic+voltammetry.">Distinguishing splanchnic nerve and chromaffin cell stimulation in mouse adrenal slices with fast-scan cyclic voltammetry.</a> American Journal of Physiology-Cell Physiology. 300 (1), 49-57 (2011).</li><li>Wolfe, J. T., Wang, H., Perez-Reyes, E., Barrett, P. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stimulation+of+recombinant+Ca(v)3.2,+T-type,+Ca(2+)+channel+currents+by+CaMKIIgamma(C).">Stimulation of recombinant Ca(v)3.2, T-type, Ca(2+) channel currents by CaMKIIgamma(C).</a> The Journal of Physiology. 538, 343-355 (2002).</li><li>Chan, S. 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=Fast+in+vivo+detection+of+myocardial+norepinephrine+levels+in+the+beating+porcine+heart.">Fast in vivo detection of myocardial norepinephrine levels in the beating porcine heart.</a> American Journal of Physiology-Heart and Circulatory Physiology. 318 (5), 1091-1099 (2020).</li><li>Chow, R. H., von R&#252;den, L., Sakmann, B., Neher, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Single-Channel Recording, Second Edition. , 245-275 (1995).</li><li>Ren, Y., 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=Facile,+high+efficiency+immobilization+of+lipase+enzyme+on+magnetic+iron+oxide+nanoparticles+via+a+biomimetic+coating.">Facile, high efficiency immobilization of lipase enzyme on magnetic iron oxide nanoparticles via a biomimetic coating.</a> BMC Biotechnology. 11, 63 (2011).</li></ol>

本方案描述了能够在 体外 和体内环境中检测和测量感兴趣的生物标志物的电容式免疫探针（CI探针）的制造和 测试 。通过将生物标志物捕获在电极尖端来实现检测。捕获事件改变了铂丝电容式免疫探针与周围导电流体环境之间的电容结，测量为响应探头中电位移位的电荷电流变化。还提出了一种独特的电采集方案，该方案允许通过静电排斥探针尖端上捕获的生物标志物，在检测周?...

用于检测和定量生物标志物的几种化学方法通常用于蛋白质化学和临床诊断，特别是在癌症诊断和心血管疾病进展评估中。目前，诸如高效液相色谱（HPLC）、酶联免疫吸附测定（ELISA）和质谱等方法依赖于通过本体流体抽取从血管区1，2，3 或通过微透析从血管区室收集样品。微透析采用已知长度的半透膜管，放置在感兴趣的区域。收集液通过管注水4 分钟以收集样品进行分析5，从而限制了时间分辨率。以这种方式，收集的样品仅提供局部微环境随时间变化的平均值，并且受到灌注速率和收集足够样品体积的限制。此外，这些方法需要汇集实验数据和信号平均;因此，他们可能无法解释受试者之间的可变性。重要的是，样本收集和随后的离线分析之间的时间排除了即时临床干预和治疗。
在本方案中，概述了使用电容式免疫探针生物传感器（CI探针）对特定生物活性肽进行时间分辨电检测。神经肽Y（NPY）从神经节后交感神经元释放，支配脉管系统，心内膜，心肌细胞和心内神经节，是心血管系统中主要的神经调节肽递质6，7，8，9。本文提出的方法旨在测量NPY，并在猪心模型中证明了实验可行性。然而，这种方法适用于任何生物活性肽，其选择性抗体是可用的10。该方法依赖于铂丝探头与功能化尖端11，12处的导电流体之间的电容结。在该应用中，相互作用通过针对靶神经肽（NPY）的抗体介导，该抗体与电极尖端结合，与导电流体环境连接。这种功能化是通过反应性聚多巴胺电沉积到铂丝探针10，13的尖端来实现的。
当抗体功能化探针放置在体内感兴趣的区域 时，诱发的内源性NPY释放导致与探针尖端上的捕获抗体结合，并且电极表面的导电液被NPY蛋白置换。电气环境中的局部变化导致具有高迁移率，高介电流体的位移，具有不动的带静电的分子。这改变了电极 - 流体界面，从而改变了其电容，其被测量为响应阶跃函数命令电位的电荷电流的变化。在每个单独的测量周期之后立即采用负的“复位”电位，通过静电相互作用排斥抗体中结合的NPY，从而清除抗体结合位点以进行后续的测量10。这有效地允许以时间分辨的方式测量NPY。独特的CI技术克服了上述基于微透析的免疫化学方法的局限性，从单个实验中测量动态生物标志物水平，而无需在多个实验上进行数据池或信号平均9，提供近乎实时的数据。此外，将这种方法适应任何感兴趣的生物标志物的能力，在时间分辨和局部尺度上存在适当的抗体，为评估疾病进展和指导治疗干预提供了免疫化学测量的重大技术进步。
用于数据采集和分析的软件是用IGOR Pro（一个完全交互式的软件环境）定制的。模数转换器（A/D）系统在计算机控制下发出命令电压，并从定制放大器获取数据。该放大器具有某些独特的功能。这些器件包括一个反馈电阻（可切换），用于四个采集通道中的每一个，允许选择1 MOhm或10 MOhm反馈电压钳位电路来集成电极的可变性。还构建了一个具有单个磁头和所有四个采集通道的相互接地/参考电路的载物台单元，以将设备放置在单个物理模块中靠近胸部的位置。使用1 MOhm反馈电阻设置来收集所有报告的数据。
滤波器和增益设置由放大器电报，并记录在数据文件中。数据通过10 kHz数字化的2极点模拟贝塞尔滤波器以1 kHz进行滤波。探头和周围导电溶液之间的电位差在探头尖端形成亥姆霍兹电容层。配体与探针尖端的抗体结合导致局部电荷改变，从而导致亥姆霍兹电容的变化。电路容性分量的这种变化导致将探头带到阶跃函数电压协议中的电位所需的注入电荷大小发生变化。因此，特定配体与功能化探头的结合导致电极电容测量的变化，作为峰值电容电流的变化。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>AgCl disc electrode</td>
			<td>Warner Instruments (Holliston, MA)</td>
			<td>64-1307</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-NPY monoclonal antibody</td>
			<td>Abcam, (Cambridge, MA)</td>
			<td>ab112473</td>
			<td></td>
		</tr>
		<tr>
			<td>Custom multichannel amplifier/ 1 M&#937; feedback resistor multichannel headstage</td>
			<td>NPI Electronic, (Tamm, Germany)</td>
			<td>NA</td>
			<td>Based on NPI VA-10M multichannel amplifier</td>
		</tr>
		<tr>
			<td>Dopamine HCl</td>
			<td>Sigma Aldrich (St. Louis, MO)</td>
			<td>H8502-10G</td>
			<td></td>
		</tr>
		<tr>
			<td>Gold-plated male connector pin</td>
			<td>AMP-TE Connectivity (Amplimite)</td>
			<td>6-66506-1</td>
			<td></td>
		</tr>
		<tr>
			<td>HEKA LIH 8+8 analog-to-digital/digital-to-analog device</td>
			<td>HEKA Elektronik, (Holliston, MA)</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Igor Pro data acquisition software, v. 7.08</td>
			<td>WaveMetrics, (Lake Oswego, OR)</td>
			<td></td>
			<td>Software driving command potential and data acquisition was custom written</td>
		</tr>
		<tr>
			<td>Masterflex L/S Standard Digital peristaltic pump</td>
			<td>Cole Palmer, (Vernon Hills, IL)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PFA-coated platinum wire</td>
			<td>A-M Systems, (Sequim, WA)</td>
			<td>773000</td>
			<td>0.005&#8221; bare diameter, 0.008&#8221; coated diameter</td>
		</tr>
		<tr>
			<td>Silicone elastomer</td>
			<td>World Precision Instruments (Sarasota, FL)</td>
			<td>SYLG184</td>
			<td></td>
		</tr>
		<tr>
			<td>Synthetic porcine NPY peptide</td>
			<td>Bachem (Torrance, CA)</td>
			<td>4011654</td>
			<td></td>
		</tr>
		<tr>
			<td>Synthetic porcine NPY peptide</td>
			<td>Bachem (Torrance, CA)</td>
			<td>4011654</td>
			<td></td>
		</tr>
	</tbody>
</table>

time-resolved in vivo measurement of neuropeptide dynamics by capacitive immunoprobe in porcine heart

在 体内 测量与疾病进展评估相关的生物标志物的能力是科学界和医学界非常感兴趣的。从当前测量某些生物标志物的方法获得的结果的分辨率可能需要几天或几周才能获得，因为它们在空间和时间上的分辨率都受到限制（例如，通过酶联免疫吸附测定[ELISA]，高效液相色谱[HPLC]或质谱分析的间质液的流体室微透析）;因此，他们对及时诊断和治疗的指导被打乱了。在本研究中，报道了通过使用电容式免疫探针生物传感器（CI探针） 在体内 检测和测量肽递质的独特技术。描述了这些探针的制造方案和 体外 表征。提供交感神经刺激诱导的神经肽Y（NPY） 在体内 释放的测量。NPY释放与去甲肾上腺素的交感神经释放相关，以供参考。数据证明了一种 在体内快速和局部测量神经肽的方法。未来的应用包括术中实时评估疾病进展和基于微创导管的这些探针部署。

电极制造和表征 制备了柔性电容式免疫探针（CI探针），并描绘了具有代表性的图像 如图1A所示。电极电位由计算机控制的电压钳位电路设置（图1B），并将电极浸入PBS中制成的聚多巴胺溶液中。将聚多巴胺电沉积到导电电极尖端13 上进行功能化。命令电位驱动探头电压，并测量注入的箝位电流。在单周期聚多巴胺沉积...

Watch this Scientific Journal Video about Time-Resolved In Vivo Measurement of Neuropeptide Dynamics by Capacitive Immunoprobe in Porcine Heart at JoVE.com

Time-Resolved In Vivo Measurement of Neuropeptide Dynamics by Capacitive Immunoprobe in Porcine Heart

已建立的免疫化学方法 在体内 测量肽递质，依靠微透析或体液抽取来获得样品进行离线分析。但是，这些都受到时空限制。本方案描述了克服现有技术局限性的电容式免疫探针生物传感器的制造和应用。

通过电容式免疫探 针在猪 心中对神经肽动力学进行时间分辨体内测量

The ability to measure biomarkers in vivo relevant to the assessment of disease progression is of great interest to the scientific and medical ...

这是首次开发出用于体内肽递质水平的时间分辨测量的方法。我们可以通过近乎实时的分析来测量单个受试者和离散位置的肽递质水平。首先，切割25厘米长的全氟烷氧基涂层铂丝。然后，使用手术刀，从一端剥离约五毫米的全氟烷氧基涂层，小心不要切入铂丝。现在，将剥离的铂线末端插入镀金的一毫米公连接器引脚中。然后，使用针鼻钳，将连接器针齿压接在铂线剥离端周围，并将铂线焊接到镀金连接器引脚上。接下来，将50毫克盐酸多巴胺加入50毫升10毫摩尔PBS中，pH 6，搅拌混合，制备多巴胺溶液。完全溶解多巴胺后，将铂丝的尖端放入含有新鲜制作的多巴胺补充PBS的容器中。将氯化银盘电极连接到头台的接地通道。然后，将氯化银圆盘放入含有多巴胺补充PBS和铂丝的容器中。在继续之前，将导线分流器连接到头级的参考通道。打开交互式数据采集软件，通过将起始电位设置为负0.6伏，结束电位设置为0.65伏，扫描速率设置为每秒0.04伏，沉积持续时间设置为420秒，设置锯齿电沉积命令电位协议。完成聚多巴胺沉积后，从容器中取出氯化银研磨颗粒和铂丝的尖端。将电线尖端放入含有pH值为7.4的PBS的微管中两到五分钟，并确保电线尖端不会接触微管的侧面或底部。然后，通过将目标抗体与pH 7.4的PBS以1：20的比例在适当尺寸的容器中组合来制备抗体溶液。接下来，将沉积在铂电极的聚多巴胺尖端在室温下在抗体溶液中两小时，确保铂丝尖端悬浮在溶液中并且不停留在微管的内表面上。将电容式免疫探针的功能尖端放入流动室中，注意不要以任何方式干扰电极尖端，因为这样做可能会损坏探头的感觉尖端。在第一次实验测试之前，执行TBS标准运行以调节电容式免疫探针，对于电压方案，将正阶跃电位设置为110毫伏，将负步进电位设置为负5毫伏，步长持续时间为20毫秒，采集持续时间设置为600秒。接下来，使用相同的TBS创建感兴趣的肽溶液以维持超级富酸盐的组成。设置一个歧管系统，其中超级富集液可以在TBS和肽补充剂TBS之间切换。使用TBS标准参数，通过将采集持续时间设置为360秒来设置肽传感数据采集协议。在聚多巴胺沉积之前，将铂丝电极的暴露尖端穿过22号皮下注射针，在针尖外留下约两毫米。使用镊子轻轻弯曲铂丝电极的尖端，形成悬挂在皮下注射针末端的倒钩。轻轻地将针头从带刺的尖端中取出，留下足够的铁丝以放置在容器中，而不会使针头接触液体，并如前所述继续进行多巴胺沉积。现在，用PBS冲洗带刺的探针尖端并将其放入抗体溶液中。然后，轻轻地从PBS上取下功能化的尖端。将皮下注射针移至倒钩电容式免疫探针，轻轻地将其植入感兴趣的区域，然后将金连接器针插入头台。植入后，取出皮下注射针头，将电极留在原位。命令电位中的正阶跃测量钳位探头电压所需的注入电流，并允许测量容性电流。负指令电位步骤通过静电排斥从抗体中清除结合的肽。阶跃函数命令波形描述了肽的时间分辨迭代检测和定量。底部迹线表示TBS中探头超融合下的命令电位和产生的电流。平衡的电容式免疫探针在六分钟内显示出较小的初始衰变和稳定的基线。神经肽Y流入流动室使电容电流超过基线。用神经肽Y抗体功能化探针测量的电容性免疫探针电流显示对低吡科莫神经肽Y敏感的浓度依赖性信号对于校准，测量所有测试浓度的标准曲线。当与负对照电容电流共绘制时，双侧星状神经节的刺激显示神经肽Y升高。在星状刺激下测量的去甲肾上腺素释放显示与神经肽Y同步释放，与诱发的交感神经反应一致。这些技术的发展可以提供心脏活动关键调节剂的术中读数，从而提供潜在的快速治疗或介入指导。这里介绍的技术是专门针对心血管部署的，但是，该技术本身适用于其他生理环境，例如尿液，骨骼肌，肾脏，脂肪组织等。

Research

JoVE Journal

Bioengineering

Time-Resolved In Vivo Measurement of Neuropeptide Dynamics by Capacitive Immunoprobe in Porcine Heart

Multiparametric Optical Mapping of the Langendorff-perfused Rabbit Heart

多参数的Langendorff灌注兔心脏的光学测绘

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

科研

生物工程

Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition

负染色的EM可视化蛋白质和大分子复合物：从电网的制备图像采集

Single particle electron microscopy (EM), of both negative stained or frozen hydrated biological samples, has become a versatile tool in structural ...

Optical Mapping of Action Potentials and Calcium Transients in the Mouse Heart

在小鼠心脏的动作电位和钙瞬变的光学测绘

The mouse heart is a popular model for cardiovascular studies due to the existence of low cost technology for genetic engineering in this species. ...

Procedure for Decellularization of Porcine Heart by Retrograde Coronary Perfusion

脱细胞猪心逆行灌注的程序

Perfusion-based  whole organ decellularization has recently gained interest in the field of  tissue engineering as a means to create site-specific ...

Millifluidics for Chemical Synthesis and Time-resolved Mechanistic Studies

Millifluidics的化学合成和时间分辨机理研究

Procedures utilizing millifluidic devices for chemical synthesis and time-resolved mechanistic studies are described by taking three examples. In the ...

Quantification of Global Diastolic Function by Kinematic Modeling-based Analysis of Transmitral Flow via the Parametrized Diastolic Filling Formalism

全球舒张功能通过静脉流的通过参数化舒张形式主义运动学建模为基础的定量分析

Quantitative cardiac function assessment remains a challenge for physiologists and clinicians.&#160;Although historically invasive methods have comprised ...

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

在组织工程由内在血管化<em&gt;在体内</em&gt;组织工程商会

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and ...

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

一个<em&gt;体外</em鼠类的椎间盘&gt;器官培养模型

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and ...

Light-sheet Fluorescence Microscopy to Capture 4-Dimensional Images of the Effects of Modulating Shear Stress on the Developing Zebrafish Heart

The hemodynamic forces experienced by the heart influence cardiac development, especially trabeculation, which forms a network of branching outgrowths ...

Rigid Embedding of Fixed and Stained, Whole, Millimeter-Scale Specimens for Section-free 3D Histology by Micro-Computed Tomography

用微计算机断层扫描法对无节段3D 组织学进行固定染色、全尺寸、毫米度试样的刚性嵌入

For over a hundred years, the histological study of tissues has been the gold standard for medical diagnosis because histology allows all cell types in ...

通过电容式免疫探 针在猪 心中对神经肽动力学进行时间分辨体内测量

通过电容式免疫探针在猪心中对神经肽动力学进行时间分辨体内测量