Name: micro-particle image velocimetry for velocity profile measurements of micro blood flows
Uploaded: 2013-04-25T12:00:00
Description: Watch this Scientific Journal Video about Micro-particle Image Velocimetry for Velocity Profile Measurements of Micro Blood Flows at JoVE.com

笔者想苏拉的阿布Mallouh和弗雷德里克·法希姆用于测试的协议，理查德·普雷沃LaVision公司的技术支持，在初始运行，她的帮助，感谢NSERC（自然科学与工程理事会，加拿大）的资金，凯瑟琳Pagiatikis和Guy Cloutier联系蒙特利尔大学的贷款Dalsa公司高速摄像机。

1。微晶片制造第一步是创建或购买你的微通道。微芯片材料有很多的选择。 选择最常见的材料是聚（二甲基硅氧烷）（PDMS）。出版物有很多PDMS加工通过软光刻16,17,18的方向。 PDMS通道一旦被制造的，也有一些表面处理，以扭转其天然疏水性。含氧等离子体处理是一种常见的选择。 周，埃利斯，Voelcker（20...

<ol>
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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Out-of-focus+effects+on+particle+image+visibility+and+correlation+in+microscopic+particle+image+velocimetry.">Out-of-focus effects on particle image visibility and correlation in microscopic particle image velocimetry.</a> Experiments in Fluids. 29, S166-S174 (2000).</li><li>Wereley, S. T., Meinhart, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+micro-particle+image+velocimetry+techniques.">Recent advances in micro-particle image velocimetry techniques.</a> Ann. Rev. Fluid Mech. 42, 557-576 (2010).</li><li>Williams, S. J., Park, C., Wereley, S. 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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flexible+methods+for+microfluidics.">Flexible methods for microfluidics.</a> Physics Today. , 42-48 (2001).</li><li>Sia, S. K., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+devices+fabricated+of+poly(dimethylsiloxane)+for+biological+studies.">Microfluidic devices fabricated of poly(dimethylsiloxane) for biological studies.</a> Electrophoresis. 24, 3563-3576 (2003).</li><li>Zhou, J., Ellis, A. V., Voelcker, N. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+developments+in+PDMS+surface+modifications+for+microfluidic+devices.">Recent developments in PDMS surface modifications for microfluidic devices.</a> Electrophoresis. 31, 2-16 (2009).</li><li>Pitts, K. L., Abu-Mallouh, S., Fenech, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contact+angle+study+of+blood+dilutions+on+common+microchip+materials.">Contact angle study of blood dilutions on common microchip materials.</a> Journal of the Mechanical Behavior of Biomedical Materials. , (2012).</li><li>Hovav, T., Yedgar, S., Manny, N., Barshtein, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alteration+of+red+blood+cell+aggregability+and+shape+during+blood+storage.">Alteration of red blood cell aggregability and shape during blood storage.</a> Transfusion. 39, 277-281 (1999).</li><li>Pitts, K. L., Mehri, R., Mavriplis, C., Fenech, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Micro-particle+image+velocimetry+measurement+of+blood+flow:+validation+and+analysis+of+data+pre-processing+and+processing+methods.">Micro-particle image velocimetry measurement of blood flow: validation and analysis of data pre-processing and processing methods.</a> Measurement Science and Technology. 23, 105302 (2012).</li><li>Kloosterman, A., Poelma, C., Westerweel, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow+rate+estimation+in+large+depth-of-field+micro-PIV.">Flow rate estimation in large depth-of-field micro-PIV.</a> Exp. Fluids. 50 (6), 1587-1599 (2011).</li><li>Goldsmith, H. L., Skalak, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemodynamics.">Hemodynamics.</a> Annual Review of Fluid Mechanics. 7, 213-247 (1975).</li></ol>

使用μPIV规模的微循环血流量测量可以洞察到了大量的有关生物医药，机械和化工过程。部分的关键因素考虑到红细胞本身，红细胞聚集或运动的荧光微粒的聚集和变形性，和在通道中的RBC的沉降密度。所有这些都可以被占，则一般遵循上述指引。有一个基本的清单，要记住，使用这个系统的良好的测量。首先，确定有多少合规系统或建议系统。为了尽量减少遵守使用油管可能在最短的和尽可能?...

人体中含有许多血管直径小于50微米，这是主要的血液和组织之间的交流网站。这些血管的血流量的研究表示一个相当大的挑战，由于规模测量的血液的流体性质。这些测量值，包括在小动脉和小静脉的压力梯度，在墙上的剪切速度分布，是与生理反应的关键因素。现在有前所未有的机遇，以解决这些测量挑战，由于新的实验技术，在微观尺度上，研究了微循环和解决这一多尺度问题。 微观粒子的图像测速（μPIV）是一个基于粒子的流量可视化技术，用于评估血液流动的速度分布在微通道通过交叉相关。 μPIV，首先圣地亚哥等人开发的，已用于与血液流变学自杉井等人的研究，在2001年使用的手法是在100μm的圆形的玻璃管1,2测量血流量。不同的方法μPIV存在。可以使用高速摄像机相关联的运动的红血细胞（红细胞），脉冲的图片可被用来示踪粒子的运动相关联。这些选项可以配上直立或倒置显微镜，根据不同的应用。在这两种情况下，其结果是一个二维速度分布图。另一种方法是使用激光共聚焦显微镜实现2D和3D的配置文件。这种方法已被应用到血液3,4,5。 微观尺度PIV有多种并发症较宏观PIV。位宏PIV中的数据可以被限制为一个单一的平面的光通过片材，但在微观尺度上卷照明是必要的。卷照明成像微血流量是一个更大的问题，如红细胞本身相比要大的channels，使用的红细胞作为示踪粒子导致的深度的相关性（DOC），它可以显着降低的准确性的互相关结果6,7,8。继Wereley 等人（1998年）为40微米高的通道与RBC作为示踪剂的DOC是8.8微米，1微米示踪粒子DOC是6.7微米。改变通道高度和放大倍率时，这种差异变得更加明显。此外，红细胞是不透明的，并在流动的红细胞的密度增加会导致成像的困难。荧光示踪粒子，在第一圣地牙哥等人（1998）所使用的，已被作为一种工具，以减少聚焦粒子的影响，倡导使用尽可能最小的颗粒。使用直径为1微米的荧光的微粒用激光耦合的一种方法，可能会降低微血流显像10的深度的焦点问题。有几个当前的状态μPIVTEC评论的hnology，其中每一个突出的重要性μPIV血流研究11,12。几个重要的考虑，必须考虑到当使用血液μPIV。在微观层面上，改善微循环规模，血液可以不被认为是均匀的流体，因为是悬挂灵活的红血细胞（红细胞），大型白细胞，血小板及其他蛋白质悬浮在一个牛顿流体（血浆） 。 的速度分布测量信号可以用来测量微血流量的某些特性。的重要因素在microhemorheology是血液的流速的速度剖面的形状，容器的壁处的剪切应力。此信息具有临床意义，，改善微循环是网站在体内的养分交换，这种交换是剪切速率依赖。有几个目前的审查研究的微循环状态的研究，以及13，14,15。 这里介绍的是μPIV血流量测量中的聚二甲基硅氧烷（PDMS）微通道的协议。按照本协议第1条中的来源渠道，在内部制造PDMS。获得认可的屠宰场猪血液样本和清理后，该协议的第2条。使用LaVision MITAS的μPIV系统，该协议第3条中所描述的所有数据。设置包括一个Nd：YAG激光（新浪潮研究，美国）和CCD照相机（图像激烈，Lavision）由一个可编程触发单元，在荧光显微镜加上3轴载物台移动中，和一台计算机控制的，除了到一个高速照相机（1M150 Dalsa公司，荷兰）溶液中加入红细胞本身的可视化。这两款相机连接到2端口光纤盒（德国蔡司定制）。在典型的在体内的血流量测量，直立的显微镜是用来跟踪的红细胞mselves，而在典型的体外应用倒置显微镜是用来跟踪示踪粒子。在这个独特的双重设置，允许两个示踪剂使用倒置显微镜成像光学框。通过高精密注射器泵的微芯片（Nexus3000，Chemyx公司，美国）被引入到血液。该系统示于图1中 ，图中的顶部部分表示为140μm，40μm的矩形通道的PDMS制作的图表，和底部代表整个系统，包括摄像机，激光，注射泵和显微镜。 电流μPIV集起坐，通常与专有软件，包括TSI，丹特克动力，LaVision。标准的交叉相关算法可以通过众多的软件选项，包括MATLAB的实现。该软件是不是关键，了解的对话方块对应以数学将成为使用ŗ好得多。在这个协议中戴维斯，LaVision的专有软件或MATLAB利用。此协议是不特定的软件，但菜单选项可能会在不同的地方，不同的软件包。

<table border="1">
	<tbody>
		<tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>poly(dimethylsiloxane) (PDMS), i.e. Sylgard-184</td>
			<td>Dow-Corning</td>
			<td>3097358-1004</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>ethylenediaminetetraacetic acid (EDTA)</td>
			<td>Sigma Aldrich</td>
			<td>E9884-100G</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>poshpate buffered saline (PBS)</td>
			<td>Sigma Aldrich</td>
			<td>P5368-10PAK</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>fluorescing micro particles</td>
			<td>Microgenics/FisherSci</td>
			<td>R900</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>glycerol (OPTIONAL)</td>
			<td>Sigma Aldrich</td>
			<td>G6279-500 ml</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>microcentrifuge, i.e. CritSpin</td>
			<td>FisherSci</td>
			<td>22-269-291</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>syringe, i.e. 50 &mu;l Gastight</td>
			<td>Hamilton</td>
			<td>80965</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>camera, i.e. Imager Intense, high speed</td>
			<td>LaVision, Dalsa</td>
			<td>Imager Intense</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>microscope, i.e. MITAS</td>
			<td>LaVision</td>
			<td>MITAS</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Nd:YAG laser</td>
			<td>New Wave Research</td>
			<td>Solo-II</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>syringe pump, i.e. Nexus3000</td>
			<td>Chemyx, Inc.</td>
			<td>Nexus-3000</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>flexible tubing, i.e. Tygon</td>
			<td>FisherSci</td>
			<td>14-169-1A</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>data processing software, i.e. DaVis</td>
			<td>LaVision</td>
			<td>DaVis</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>centrifuge, i.e. Thermo Scientific CL2</td>
			<td>Thermo Scientific</td>
			<td>004260F</td>
			<td>&nbsp;</td>
		</tr>
	</tbody>
</table>

micro-particle image velocimetry for velocity profile measurements of micro blood flows

微观粒子图像测速（μPIV）是用于可视化成对的图片，接种在血液流动的微颗粒。图像互相关，给出一个准确的流速剖面。一个协议为μPIV测量血流量在微。规模的微循环，血液不能被认为是一种均匀的流体，因为它是一个灵活的粒子悬浮液悬浮在血浆中，牛顿流体。从这些测量中，可以推导出剪切速率下，最大速度，速度分布图的形状，和流速。如震源深度，粒子浓度，系统符合的几个关键参数，随着各种血细胞比容和流量条件下的实施例和代表性的结果，以确保准确的，有用的数据。

在所有的数字中，流是左到右的原始图像，并在计算速度剖面向上。与血液血细胞比容H = 10以10微升/小时的流动获得的原始数据，例如， 如图2所示。上游的数据可能会被交叉没有任何的数据处理，以实现速度分布密切相关。皮茨， 等人，（2012B）的前处理和数据处理方法的影响进行了讨论。 = 10和10微升/小时的流速得到的速度分布图，从与图2相似在hematorcrit H的数...

Watch this Scientific Journal Video about Micro-particle Image Velocimetry for Velocity Profile Measurements of Micro Blood Flows at JoVE.com

Micro-particle Image Velocimetry for Velocity Profile Measurements of Micro Blood Flows

微观粒子图像测速（μPIV）是用来配对接种于血流量，给出一个准确的流速剖面互相关微颗粒的图像可视化。剪切速率下，最大速度，速度分布图的形状，和流速，每个都具有临床应用，可以是来自于这些测量。

微观粒子图像测速技术，微血液流动的速度剖面测量

Micro-particle image velocimetry (&mu;PIV) is used to visualize paired images of micro particles seeded in blood flows. The images are cross-correlated to ...

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A Versatile Automated Platform for Micro-scale Cell Stimulation Experiments

一种多功能自动化平台微尺度细胞刺激实验

Study of cells in culture (in vitro analysis) has provided important insight into complex biological systems. Conventional methods and equipment for in ...

Improved Method for the Preparation of a Human Cell-based, Contact Model of the Blood-Brain Barrier

改进方法对血脑屏障的人体细胞为主，接触模型的制备

The blood-brain barrier (BBB) comprises impermeable but adaptable brain capillaries which tightly control the brain environment. Failure of the BBB has ...

Hybrid &#181;CT-FMT imaging and image analysis

Hybrid Ãƒâ€šÃ‚ÂµCT-FMT imaging and image analysis

混合μCT-FMT成像和图像分析

Fluorescence-mediated tomography (FMT) enables longitudinal and quantitative determination of the fluorescence distribution in vivo and can be used to ...

Microfluidic Buffer Exchange for Interference-free Micro/Nanoparticle Cell Engineering

微流控缓冲液更换为无干扰微/纳米颗粒细胞工程

Engineering cells with active-ingredient-loaded micro/nanoparticles (NPs) is becoming an increasingly popular method to enhance native therapeutic ...

Perfusable Vascular Network with a Tissue Model in a Microfluidic Device

微流控装置中组织模型的 Perfusable 血管网

A spheroid (a multicellular aggregate) is regarded as a good model of living tissues in the human body. Despite the significant advancement in the ...

Biological Compatibility Profile on Biomaterials for Bone Regeneration

骨再生生物材料的生物相容性曲线

Large non-union bone fractures are a significant challenge in orthopedic surgery. Although auto- and allogeneic bone grafts are excellent for healing such ...

Meso-Scale Particle Image Velocimetry Studies of Neurovascular Flows In Vitro

Meso-Scale Particle Image Velocimetry Studies of Neurovascular Flows In Vitro

神经血管流动的中尺度粒子图像测速研究

Particle image velocimetry (PIV) is used in a wide variety of fields, due to the opportunity it provides for precisely visualizing and quantifying flows ...