Name: the ex vivo isolated skeletal microvessel preparation for investigation of vascular reactivity
Uploaded: 2012-04-28T14:00:00
Description: Watch this Scientific Journal Video about The ex vivo Isolated Skeletal Microvessel Preparation for Investigation of Vascular Reactivity at JoVE.com

这项工作是由美国心脏协会（EIA 0740129N）和NIH的T32 HL90610支持。

1。实验前<ol><li>实验一天前，站适当尺寸的玻璃毛细管拉成微电极（无论是水平或垂直的车夫，都可以使用）。尖端直径可以根据被隔离的船只随时调整，虽然我们通常使用直径范围50-150微米之间。这些微血管站了以下丁烷火焰加热到适当的配置，然后弯曲。微量提示身体打破近似直径（用细镊子）微血管中的问题，可以适当的构象，如针对特定应用所需的研磨或抛光。读者对这些程序的广泛?...

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T., Frazer, D., Castranova, V., Nurkiewicz, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoparticle+inhalation+impairs+endothelium-dependent+vasodilation+in+subepicardial+arterioles.">Nanoparticle inhalation impairs endothelium-dependent vasodilation in subepicardial arterioles.</a> J. Toxicol. Environ. Health A. 72, 1576-1584 (2009).</li><li>Jernigan, N. L., LaMarca, B., Speed, J., Galmiche, L., Granger, J. P., Drummond, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dietary+salt+enhances+benzamil-sensitive+component+of+myogenic+constriction+in+mesenteric+arteries.">Dietary salt enhances benzamil-sensitive component of myogenic constriction in mesenteric arteries.</a> Am. J. Physiol. Heart Circ. Physiol. 294, H409-H420 (2008).</li><li>Stapleton, P. A., Goodwill, A. G., James, M. E., Frisbee, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+mechanisms+of+endothelium-dependent+dilation+in+skeletal+muscle+arterioles+with+genetic+hypercholesterolemia.">Altered mechanisms of endothelium-dependent dilation in skeletal muscle arterioles with genetic hypercholesterolemia.</a> Am. J. Physiol. Regul. Integr. Comp. Physiol. 293, R1110-R1119 (2007).</li><li>Goodwill, A. G., Stapleton, P. A., James, M. E., d'Audiffret, A. C., Frisbee, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+arachidonic+acid-induced+thromboxane+generation+impairs+skeletal+muscle+arteriolar+dilation+with+genetic+dyslipidemia.">Increased arachidonic acid-induced thromboxane generation impairs skeletal muscle arteriolar dilation with genetic dyslipidemia.</a> Microcirculation. 15, 621-631 (2008).</li><li>Baumbach, G. L., Hadju, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanics+and+composition+of+cerebral+arterioles+in+renal+and+spontaneously+hypertensive+rats.">Mechanics and composition of cerebral arterioles in renal and spontaneously hypertensive rats.</a> Hypertension. 21, 816-826 (1993).</li><li>Uchida, E., Bohr, D. F., Hoobler, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+studying+isolated+resistance+vessel+from+rabbit+mesentery+and+brain+and+their+responses+to+drugs.">A method for studying isolated resistance vessel from rabbit mesentery and brain and their responses to drugs.</a> Circ. Res. 4, 525-536 (1967).</li><li>Davis, M. J., Kuo, L., Chilian, W. M., Muller, J. M. I. s. o. l. a. t. e. d., Barker, J. H., Anderson, G. L., Menger, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+23.">Chapter 23.</a> Isolated, perfused microvessels. In: Clinically Applied Microcirculation Research. 32, 435-456 (1995).</li><li>Lombard, J. H., Liu, Y., Fredricks, K. T., Bizub, D. M., Roman, R. J., Rusch, N. 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+and+mechanical+responses+of+rat+middle+cerebral+arterieal+to+reduced+PO2+and+prostacyclin.">Electrical and mechanical responses of rat middle cerebral arterieal to reduced PO2 and prostacyclin.</a> Am. J. Physiol. 276, H509-H516 (1994).</li></ol>

提出的协议描述的骨骼肌微血管隔离，清除和双插管，虽然这一般技术可以很容易地适用于大多数组织。对于当前的手稿，“动脉”一词已被用于由作者来形容一个阻力血管直径在70-120微米休息积极的基调下，这也是一个主要因素调节器官灌注阻力或之间不等组织。 一些修改，这个系统可以安装多个应用程序的具体调查，并随时可以提供更深入的实验（例如，17膜电?...

<table border="1"><tbody><tr><td> 试剂和设备 </td><td> 公司 </td><td> 评论/目录＃ </td></tr><tr><td>船厅</td><td>习俗</td><td>戴夫Eick（都城） </td></tr><tr><td>加热循环水浴</td><td> PolyScience和哈克</td><td>哈克直流10 </td></tr><tr><td>滴管</td><td>冯检基Haer公司</td><td>毛细管2.0 mm外径×1.0 mm内径（27-33-1） </td></tr><tr><td>血压计</td><td>世界精密仪器</td><td></td></tr><tr><td>水夹套水库的</td><td>习俗</td><td></td></tr><tr><td>外部光源</td><td>世界精密仪器</td><td> NovaFlex经股</td></tr><tr><td>吸取普勒</td><td>微观仪器</td><td> PMP102 Micropipet普勒</td></tr><tr><td>全部完井MENT的手术工具</td><td>罚款科学工具</td><td>杜蒙</td></tr><tr><td>超精细镊子</td><td>罚款科学工具</td><td> INOX＃5 </td></tr><tr><td>丝绸缝合线</td><td> ethilon </td><td> ＃10-0或9-0 </td></tr><tr><td>立体显微镜</td><td>奥林巴斯</td><td>奥林巴斯SZ-11 </td></tr><tr><td>模拟视频卡钳</td><td> boeckeler </td><td>通过控制器（VIA-100） </td></tr><tr><td>高分辨率模拟摄像机</td><td>松下</td><td> GP - MF 602 </td></tr><tr><td>氧气罐</td><td>区域性</td><td>余额21％的氮和5％CO 2的平衡氮</td></tr><tr><td>管道</td><td>聚乙烯</td><td></td></tr><tr><td>排水泵</td><td>科尔帕默仪器有限公司</td><td></td></tr><tr><td>修饰大鼠的PSS </td><td>看到配方如下</td><td></TD&gt; </tr><tr><td>凡Breemen松弛的PSS </td><td>看到配方如下</td><td></td></tr></tbody></table> 表1。隔离的微血管站提出的数字设置的主要组成部分的清单。 <table border="1"><tbody><tr><td> 修改大鼠的PSS食谱 </td><td> 为了使两公升的PSS </td><td> 20X盐库存（2L） </td><td> 20X缓冲库存（2L） </td></tr><tr><td>氯化钠</td><td></td><td> 278.0Ğ </td><td></td></tr><tr><td>氯化钾</td><td></td><td> 14.0Ğ </td><td></td></tr><tr><td>硫酸镁4-7H 2 O的 </td><td></td><td> 11.5Ğ </td><td></td></tr><tr><td>氯化钙2-H 2 O </td><td></td><td> 9.4Ğ </td><td></td></tr><tr> &lt;TD&gt;碳酸氢钠3 </td><td></td><td></td><td> 80.8Ğ </td></tr><tr><td> EDTA的</td><td></td><td></td><td> 0.4Ğ </td></tr><tr><td>的NaH 2 PO 4计 </td><td> 0.28Ğ </td><td></td><td></td></tr><tr><td>葡萄糖</td><td> 1.98Ğ </td><td></td><td></td></tr><tr><td> 20倍的盐库存</td><td> 100毫升</td><td></td><td></td></tr><tr><td> 20X缓冲库存</td><td> 100毫升</td><td></td><td></td></tr><tr><td>蒸馏水</td><td> 1800毫升</td><td></td><td></td></tr></tbody></table> 表2。配方为标准生理盐溶液中用于孤立微血管协议“（PSS）。 食谱上的评论：2公升盐库存和缓冲库存的大号2。这些C冷藏不使用时，但动摇他们之前往往准备的PSS。额外的成分，添加在最终的PSS的准备时间。 <table border="1"><tbody><tr><td> 凡Breemen松弛的PSS </td><td> 为了使2升的PSS </td><td> 20X盐库存（1L） </td><td> 20X缓冲库存（1L） </td></tr><tr><td>氯化钠</td><td></td><td> 107.4Ğ </td><td></td></tr><tr><td>氯化钾</td><td></td><td> 7.0Ğ </td><td></td></tr><tr><td>硫酸镁4-7H 2 O的 </td><td></td><td> 5.76Ğ </td><td></td></tr><tr><td>氯化镁2-6H 2 </td><td></td><td> 81.32Ğ </td><td></td></tr><tr><td> 碳酸氢钠 </td><td></td><td></td><td> 40.4Ğ </td></tr><tr><td>美东时间一</td><td></td><td></td><td> 0.2Ğ </td></tr><tr><td> EGTA </td><td></td><td></td><td> 15.22 </td></tr><tr><td>的NaH 2 PO 4计 </td><td> 0.28Ğ </td><td></td><td></td></tr><tr><td>葡萄糖</td><td> 1.98Ğ </td><td></td><td></td></tr><tr><td> 20倍的盐库存</td><td> 100毫升</td><td></td><td></td></tr><tr><td> 20X缓冲库存</td><td> 100毫升</td><td></td><td></td></tr><tr><td>蒸馏水</td><td> 1800毫升</td><td></td><td></td></tr></tbody></table> 表3。配方范Breemen松弛生理盐溶液（PSS），在孤立的微血管协议的零积极音的条件下使用。 食谱上的评论：1 L一盐库存ND 1升的缓冲库存。不使用时，可以冷藏，但动摇他们之前往往准备的PSS。额外的成分，添加在最后松弛的PSS的准备时间。

the ex vivo isolated skeletal microvessel preparation for investigation of vascular reactivity

隔离的微血管准备是体外制备，允许考试因素的控制血管直径的不同贡献，因此，灌注阻力1-5。这是一个经典的实验准备，在很大程度上，最初是由内田等 15，几十年以前。这最初的描述提供了进行了广泛的修改和提高，主要是在实验室的布赖恩博士杜陵在弗吉尼亚6-8大学，技术的基础上，我们提出了电流的方法，在下面的网页。这种准备，具体是指在大鼠作为首选微血管纤细的动脉，但基本的准备，可以很容易地应用到几乎任何其他组织或器官9-13跨物种隔离的船只。机械（即二维）在孤立的微血管的变化，可以很容易地评估在浩如烟海的生理（例如，缺氧，血管内的压力，或剪切）或药理的挑战，并能提供洞察到机械元素组成一个完整的综合措施，虽然体外 ，组织响应。这种方法的意义是，它允许简便操作的影响微血管直径的综合调控，同时也让许多从其他来源，包括血管内压力（生肌），自主支配，血流动力学的贡献控制（例如，剪切应力），内皮依赖或独立的刺激，激素，实质影响，提供了部分清单。在适当的实验条件和适当的目标，这可以作为一个优势在体内或原位组织/器官的筹备工作，不轻易让浅显的控制更广泛的系统性变量。 马约旦基本上是该制剂的限制其优势的结果。根据定义，这些船只的行为被许多最重要的贡献者调节血管阻力已被删除，包括神经，体液免疫，代谢，因此等的条件下，研究，研究者应注意避免过度所收集的数据利用这个准备的解释和推断。该制剂方面关心的其他重要领域，它可以很容易损坏，如内皮衬里或血管平滑肌细胞成分，如变量的错误源可以被引入。强烈建议，个别研究者利用，以确保该制剂的质量，无论是在实验开始，并定期在整个协议的过程中适当的测量。

Watch this Scientific Journal Video about The ex vivo Isolated Skeletal Microvessel Preparation for Investigation of Vascular Reactivity at JoVE.com

The ex vivo Isolated Skeletal Microvessel Preparation for Investigation of Vascular Reactivity

一个<em&gt;体外</em&gt;编制说明两个血管反应性血管活性物质的刺激和通过被动墙力学的基本结构性能评估的审讯股薄肌的最大阻力小动脉的隔离。

“体外骨骼肌微血管制备离体血管反应性的调查

The isolated  microvessel preparation is an ex vivo preparation that allows for examination of the different contributions of  factors that control vessel ...

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The ex vivo Isolated Skeletal Microvessel Preparation for Investigation of Vascular Reactivity

In vivo Micro-circulation Measurement in Skeletal Muscle by Intra-vital Microscopy

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BACKGROUND: Regulatory factors and detailed physiology of in vivo microcirculation have remained not fully clarified after many different modalities of ...

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Described here is a method to measure contractility of isolated skeletal muscles. Parameters such as muscle force, muscle power, contractile kinetics, ...

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A Novel Ex vivo Culture Method for the Embryonic Mouse Heart

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Developmental studies in the mouse are hampered  by the inaccessibility of the embryo during gestation. Thus, protocols to  isolate and culture individual ...

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The isolated blood-perfused lung preparation is widely used to visualize and define signaling in single microvessels. By coupling this preparation with ...

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While the isolation and culture of vascular smooth muscle cells (VSMCs) from large vessels is well established, we sought to isolate and culture VSMCs ...