这项工作是由授权号码0953652来自美国国家科学基金会和部分由乔治·梅森大学图书馆开放获取出版基金的部分资助。我们要感谢约翰·罗伯特·克雷斯曼金博士提供接入高速摄像机。

1。仪器仪表矢量TDI是基于估算从两个或多个独立的方向取多普勒速度测量得到的速度矢量。超声系统具有界面研究用于开发VTDI。该研究的接口允许水平低的波束形成和脉冲序列控制使用的软件开发工具包（SDK）。 5-14 MHz的线阵探头，由128换能器元件，并与视图一个38毫米的领域被使用。研究接口被用来将阵列传感器分成两个发射和接收孔，并通过与相对于法线15°转向的接收波束。发射波束集中在感兴趣的区域（ 如腹部肌肉）。发送和接收孔径分别设置为32个元素。 八个科目，4男4女（29.7±6.5岁）被招募了这项研究。从该对象运动的措施右下肢被使用八位相机运动捕捉系统具有高速性能和200Hz的采样率捕获。在实验过程中地面反作用力的数据通过两个力板采样在2000赫兹获得。 一个高速相机安装在三脚架上，并放置在离被摄对象2 M，用于捕获下降着陆在500帧/秒。 2。主题准备<ol><li>要求受试者穿一条短裤，运动内衣或短T恤和跑鞋。 </li><li>指导受试者进行10分钟自导热身和拉伸前的数据集合。这是为了避免不正常的肌肉收缩，并减少任何肌肉痉挛的范围。 </li><li>在热身环节后，将反光标记对身体特定的地标。具体而言，在更大trochanters，双侧内侧和外侧膝处校准标记和内侧和雷特拉升踝关节。放置跟踪标记上的后部和髂前上嵴和地方集群上的每个脚19-20大腿和小腿，并且五种标志物。 </li><li>指导受试者的立场的3D摄像机的对焦区域中心获得一个静态试验。与会者必须站在测力板与他们的武器在他们的肩膀上，获得静态3D动作捕捉数据。 </li><li>然后，将超声换能器中的换能器保持器，并确保良好的玩意儿，避免从换能器保持器的超声换能器的脱离。换能器支架用LEXEN聚碳酸酯和可模制的塑料制成。 </li><li>为了确保与皮肤和超声换能器接触良好，适用于超声换能器发射凝胶大方量。 </li><li>将超声换能器一起就此事向图像的大腿传感器支架股直肌在纵向斧是。该传感器必须放置在半路前髂前上棘与横向epicondoyle图像直肌腹部肌肉的股二头肌之间。前固定超声换能器和换能器持有人的腿，得到股四头肌肌肉群的轴向切片。以此为指导，确保超声换能器现在成像股直肌和不动更多的外侧或内侧，以避免成像vastii肌肉群。 </li><li>现在，使用一个有凝聚力的自粘绷带到传感器​​支架固定到主体的大腿。让这个程序上的步骤不阻止或覆盖反射标记。自粘绷带不能松懈或过于紧张。包扎不严将风险超声换能器落在下拉登陆任务中和过度紧张的包扎会引起不适，浅表性破坏血流量，并可能改变下降着陆动态。 </li><li>要将它放在他的高速相机至少有2米的距离在矢状面的主题，收集视频在500帧/秒。对焦相机镜头，以确保受试者的整个下降着陆序列可被抓获。 </li></ol> 3。实验协议<ol><li>一旦所有的标记和超声换能器是安全的，要求受试者站在身高30厘米地方的平台，在50厘米的测力板。确保周围的平台（约2.5米）的面积显然会妨碍下降着陆任务或伤害问题的任何对象。这包括超声换能器线。 </li><li>指示对象把他们的手放在臀部前开始下降着陆任务和整个下降着陆序列。 </li><li>启动数据收集超声，三维运动捕捉，力板，并开始下降着陆任务之前的高速摄像机。不同的仪器之间的同步功能，可以实现ð通过使用一个单一的按键来启动所有的数据采集。连接到键盘的压力传感器可以用于产生当一个特定的键被按下时的同步触发信号。 </li><li>直接受到从平台和土地用两条腿进行降着陆任务，同时进行。确保受试者从箱子掉落，而不是从它跳跃。提供有关着陆技术没有具体说明。 </li><li>停止收集数据，一旦问题已完全稳定，并完成了降着陆序列。 </li><li>重复此协议每科五次。 </li></ol> 4。超声数据分析<ol><li>出口和从超声波系统的原始数据存储到计算机上。 </li><li>从各原射频（RF）的超声数据接收光束被数字化，在40兆赫。利用MATLAB处理数据。 </li><li>对RF数据进行正交解调，以除去所述载波频率。删除统计ionary和低频杂波通过使用20赫兹的高通滤波器的滤波从每个接收波束和每个深度的正交数据。 </li><li>估计沿两个速度接收使用传统的自相关速度估计器21的光束。 </li><li>结合各个速度波形，以获得横向（沿换能器）和轴向（垂直于换能器）的速度波形的整个下降着陆序列， 如图1所示。 </li><li>取得所得的速度向量的大小从利用公式1如先前所描述的22个人的速度分量: <img alt="式（1）" src="/files/ftp_upload/50595/50595equation1.png" /> 其中，β为光束转向角，f 1和f 2分别是两个接收到的频率分量和f t为发射频率。 </li><li>计算横向和轴向应变速率解/ dt的使用空间梯度已废除的横向和轴向速度。 <img alt="公式2" src="/files/ftp_upload/50595/50595equation2.png" /> 其中V 2和V1 顷估计在由距离L隔开的两个空间位置的瞬时速度</li><li>计算轴向和侧向应变，E，由轴向和横向应变率分别整合。 <img alt="式（3）" src="/files/ftp_upload/50595/50595equation3.png" /></li></ol> 5。三维运动捕捉数据分析<ol><li>三维运动捕捉数据导出到计算机作进一步分析。 </li><li>使用静态站立试验中，创建一个使用3D动作捕捉软件用最小二乘优化23运动学模型（骨盆，大腿，小腿和足部）。 </li><li>使用此运动学模型来量化运动在髋关节，膝关节和踝关节。 </li><li>过滤器使用的是四阶低通Butterwor的反射标记轨迹和地面反作用力个滤波器的7赫兹和25赫​​兹，分别使用3D动作捕捉软件的截止频率。 </li><li>计算的3-D接头的力和力矩使用标准的逆动力学分析，采用估计每个参与者按照Dempster组合方法段惯性特征的运动学和地面部队的数据。分部间关节力矩被定义为内部的时刻（ 例如膝盖内部分机时刻将抵制应用于膝关节屈曲载荷）。 </li></ol> 6。高速摄像机数据分析<ol><li>从高速相机数据导出视频到电脑进行分析，并与超声和三维运动捕捉运动数据进行比较。 </li><li>播放电影15帧/秒，并观察降着陆动态。 </li><li>然后，量化的换能器支架的运动和超声波换能器的整个滴着陆试验通过跟踪可见标记的anatomi中的位移使用高速的视频数据CAL标志性建筑。评估降着陆动态，也可以同时进行，以便更好地了解不同的发射和着陆的风格。 </li></ol>

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没有作者的任何财务披露或利益与冲突研究批准本机构的内部评级法。

超声显像必须提供肌肉运动的直接评估，动态的研究，可以补充常规的措施，如3D动作捕捉，测功，肌电图，和地面反作用力测量的能力。这种方法可以广泛地适用于基本的生物力学研究与临床评价。主要有三种方法使用超声估计组织运动:（1）使用互相关的原始射频（RF）超声数据或包络检波灰度（或B模式）的图像数据斑点跟踪方法。这些技术已被广泛用于在两个骨骼24-25和心脏26肌肉运动跟踪和估算，（2）用于跟踪肌肉束状或特征27-28和（3）在两个心脏29中使用组织多普勒成像技术的图像处理方法-30和骨骼31运动估计。基于空间交叉-C斑点追踪orrelation已被广泛地用于跟踪组织的运动，并且可以具有子像素分辨率的跟踪运动。然而，散斑图中较大的去相关议案很快。运动出了图像平面的也构成为斑点追踪的一个挑战。方法跟踪肌束长度有更好的适用性所在的整个分册的动态任务中可视化的形象。依赖于处理图像数据的方法具有有限的成像帧率较低的时间分辨率，因此在高的速度无法跟踪运动。此外，这些肌束跟踪方法是向平面外的运动非常敏感。从而探针运动相对于肌肉可能会导致跟踪失败。从传统的组织多普勒成像（TDI）速度估计可以有更高的时间分辨率，以及更强大的小探头的动作。多普勒方法可以估算的速度分量仅沿超声束，从而多普勒估计可能是不准确的杜E要声波作用的不同角度与肌肉的运动。我们提出的VTDI方法通过利用转向以不同的角度两种不同的超声波束克服了这个问题，因此速度估计是独立于成像平面的声波作用的角度。此外，VTDI的有效时间分辨率可以约0.1毫秒，因此这种方法可以在动态活动跟踪骨骼肌的运动（ 如降着陆，步态和跑步）。 我们的方法的其他优点包括使用基于用于执行矢量组织多普勒成像临床超声系统的线性阵列的成像传感器。我们的电控发射/接收波束控制，光圈的大小和焦点的位置，用于扫描的大视场。此外，这种方法可以扩展到执行双面VTDI和同步实时成像。我们的系统还允许我们进行常规B模式成像升ocate感兴趣的区域的组织应变和运动学定量。由于这种方法在临床上的扫描仪实现，我们已经能够在一个步态实验室进行生物力学研究部署此VTDI方法。 这种技术的局限性必须承认。各种因素影响多普勒测量的准确性。在2维（沿与横过肌纤维）VTDI基于速度估计需要的线性阵列换能器被分成两个发射/接收子孔径（32个元素宽），并通过15°转向的光束。操纵超声发射和接收波束，以更高的角度会影响由于栅瓣速度的措施。此外，光束的重叠区域中VTDI的区域具有不同的光束聚焦改变深度32，有可能影响速度的估计。多普勒估计的方差取决于吨（1）在分析的时间范围内加速和组织的减速（2）方差多普勒范围内浇口速度的问题（3）用于宽带的孔径内的不同多普勒角谱法发送和接收的超声波束，也被称为几何扩大33和发射的超声波脉冲（4）的带宽，由于多普勒频移正比于载频34。几种方法可以被用于限制的方差。根据相速度估计器，如自相关，通常利用相比谱估计器更小的分析时间窗，但他们估计平均多普勒频移，而不是峰值偏移。宽带频谱估计类似的2D傅立叶变换35能减少方差由于脉冲带宽。在VTDI，它采用了两个转向多普勒束的情况下，组织速度的光束重叠区域相对于该肌肉的中的方差是另一个要考虑的因素。 股直肌肌肉收缩是在3D和收缩veloc性在空间上变化沿肌肉。因此，要慎重选择该地区的利益是很重要的。 在这项研究中，我们在使用VTDI八个健康志愿者下 ​​降着陆任务调查的股直肌肌肉运动的重复性。尽管试验是独立的，我们观察到高度相关的和可重复的峰值收缩速度进行试验之间的个人。目前，我们正在招募在我们的研究更多科目，进一步研究这种模式。这项研究提供了非侵入性和腹直肌的收缩速度的实时测量过程中忽略着陆股二头肌肌肉。下降着陆任务（ 图2）的各个阶段，观察收缩速度以下模式:1。肌肉的收缩速度的膝关节屈曲（发射阶段）和延伸期间支配在横向方向相对于轴线方向（在这方面的风PHASE）。这是意料之中的，因为股直肌肌肉时期间在空中阶段发射阶段和向心收缩进行离心收缩。 2。在第三阶段（脚趾触地），用小到可以忽略轴向肌肉速度低外侧肌肉速度。这相当于在这个阶段3，以降低股直肌肌肉收缩。脚后跟接触地面刚过大幅增加轴向和横向肌肉速度。这可能是由于肌肉的形状由于压缩经历两个偏心收缩和变化，从而导致增加沿着肌肉纤维和正常的肌肉纤维，分别速度。尽管事实是下降着陆任务是一种高冲击任务，VTDI证明重复股直肌速度。这种超声技术能有临床意义，因为这肌肉，主要负责保护膝关节的过度负荷。因此，患者的ACL重建股直肌肌肉的进一步评估是必要的了解，导致早期和发病加速OA的机制。 虽然在本研究的参与者都要求从30厘米高的平台上执行一个自然下降着陆任务，我们发现在跳跃或发射的高度差异。此外，使用高速摄像机数据，有人指出，所有科目都有不同的下降着陆的风格。这可以解释在腹直肌的峰值得到的速度值主体之间的细微差别股二头肌肌肉作为任务期间在激活模式可能存在的差异造成的。另一可能的因素是在股直肌肌肉，这可能导致不同程度的肌肉收缩，并迫使生产截面积的差异。

肌肉骨骼疾病是广泛流行在成年1。他们是在美国2领先的慢性疾病，并呈报影响全世界3人25％。肌肉骨骼疾病都与日常生活能力（ADL），功能限制和生活4质量较低的活动功能下降有关。他们的经济负担，是因为失去了工作效率和高医疗费用4显著。几个这些疾病的病理生理机制仍然不充分的理解。例如，骨关节炎（OA）4下重建前交叉韧带（ACL）损伤的发病机制已被链接到改建股四头肌肌力和功能5，但其机制尚不清楚。为了阐明其机制，有必要更好地了解动态肌肉功能。 功能个别肌肉的评估，部分或整个任务涉及到日常生活和积极的生活方式（ 如运动）的性能时可以提供约肌功能及其在这些疾病的发病机制和病理生理作用的潜力进一步了解。另外肌肉功能改善的康复过程中的量化可以作为衡量的结果。测量肌肉和关节功能在临床上的常规技术包括身体检查，如运动范围，肌力和/或肌肉群的耐力。目前在临床上，肌电图（EMG）来评估肌肉激活/助活化，频率和肌肉活动的振幅。然而，肌电图是在肌肉电活动的措施，并不一定提供有关的肌肉力量，收缩能力和肌肉的功能等因素的信息。其他复杂的生物力学评估，如三维运动捕捉系统F或关节动力学和运动学和力板的地面反作用力可以在步态实验室6-9进行。由这些技术所进行的测量是在关节水平，动态或功能性活动过程中不一定能直接了解个别肌肉功能。同时进行肌肉的成像，同时进行动态活动的能力，可能会导致在肌水平更好和更逼真的功能评估。 大多数研究都在静态俯卧位集中在肌肉功能，并且这种方法可以开辟新的途径，进一步加强在实时情况下，我们的肌肉行为的理解。 超声诊断可以使肌肉和肌腱的直接成像，实时的，因此是日常生活中测量肌肉骨骼动力学和功能有吸引力的选择。超声为基础的定量措施肌肉形态学和体系结构，如肌肉厚度，长度，宽度，横截面面积（CSA），纤维羽状角和肌束长度已被广泛使用10-12。近年来，图像处理的方法已被采用时的动态任务13-14评估和量化这些定量度量。这些进步使体内的肌肉功能的新方法学角度来理解。然而，这些方法都主要依赖于使用传统的灰度（或B型），超声成像，并因此还没有完全利用超声波的可能性，以测量组织速度，应变和利用多普勒原理应变率，已被证明是有价值的在评价心肌功能15-16。 我们已经开发出一种载体组织多普勒成像（VTDI）技术，可以测量收缩速度，应变和应变率与高时间分辨率（子millisecond）在动态活动17-18。具体地说，VTDI技术可以在高度动态的任务（ 例如，下拉式着陆，步态等 ）以高帧频使肌肉和肌腱的测量。该VTDI技术是一种改进了传统多普勒超声，它估算沿超声波束的速度的唯一组分，并因此依赖于声波作用角度。 VTDI估计使用转向以不同的角度两个不同的超声波束的肌肉和腱的速度，因此是独立于成像平面的声波作用的角度。然而，由于肌肉的收缩发生在三维中，成像平面的角度仍然是重要的。我们已经实施这种方法对市售的超声系统具有界面的研究，使这些测量在临床上作出。 调查重复性和VTDI系统·潜在的适用性时间在过程中的动态任务测量股直肌肌肉速度，我们进行了对健康成人志愿者进行了初步研究。本文演示的方法和实验装置估算的收缩速度，应变和腹直肌的应变率在下降着陆任务， 股二头肌肌肉与亚毫秒级的时间分辨率。

<table border="1">
		<tbody>
		 <tr>
			<td>Name of Equipment</td>
			<td>Company</td>
			<td>Model Name</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Ultrasound System</td>
			<td>Ultrasonix</td>
			<td>Sonix RP</td>
			<td></td>
		 </tr>
		 <tr>
			<td>3D Motion Capture System</td>
			<td>Vicon Motion Systems</td>
			<td>Vicon T-20</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Force Plates</td>
			<td>Bertec Corporation</td>
			<td>Bertec 4060-10</td>
			<td></td>
		 </tr>
		 <tr>
			<td>High Speed Camera</td>
			<td>Photron</td>
			<td>Photron 512 PCI 32K</td>
			<td></td>
		 </tr>
		</tbody>
</table>

a novel application of musculoskeletal ultrasound imaging

超声检查是一个有吸引力的方式，在动态影像的任务肌肉和肌腱的运动，并能提供生物力学研究互补方式方法在临床或实验室设置。为了实现这一目标，从超声图像肌肉运动的定量方法是基于图像处理的正在开发中。这些方法的时间分辨率通常是不足够的高度动态的任务，比如下拉式菜单着陆。我们建议，利用量化的肌肉运动的多普勒方法的新方法。我们已经开发了一种新的矢量组织多普勒成像可用于测量肌肉骨骼收缩速度，应变和应变率与期间使用超声动态活动亚毫秒级的时间分辨率（VTDI）技术。这一初步研究的目的是调查的重复性和VTDI技术的潜在应用在测量肌肉骨骼veloc在下降着陆任务伊蒂埃斯，在健康受试者。该VTDI测量可以同时与其它生物力学技术，如3D动作捕捉关节运动学和动力学，肌电图肌肉活化和测力板的地面反作用力的时间来执行。这些互补的技术整合可能会导致更好地理解动态肌肉功能和肌肉骨骼功能障碍疾病的发病机制和病理生理基础。

从我们以前的工作表明了方法的代表性结果介绍如下。而在我们目前的研究使用的方法整合成像和动作捕捉，下面提出的有代表性的结果是从那里分别进行这些测量的研究。 一，超声（VTDI） 使用从3D动作捕捉和高速摄影机，拍摄对象的跳转的模式的数据，着陆和稳定阶段进行了研究每个审判。轴向和横向股直肌的速度从VTDI进行了比较，从3D动作捕捉和高速摄像机采集到的数据。使用此数据，在整个降着陆序列的轴向和横向股直肌肌肉速度的时间特性进行了研究。正面横向速度对应股直肌肌肉离心收缩时膝关节屈曲，而在膝关节伸直负侧向速度对应的肌肉向心收缩。这示于图2。所有受试者在整个下降着陆序列历时约1.45±0.27秒。 对于每个受试者，在轴向和横向肌肉速度表明试验之间有很强的可重复性为0.99的斜率和R2 = 0.75（ 图3）。六出八个科目速度值是在一个类似的范围48-62厘米/秒，而二级学科（均为男性）有更高的速度。男性（72.96厘米/秒）呈现比女性显著高于肌肉速度（48.71厘米/秒），P = 0.029，调整为每个主题的个性化体重和肌肉厚度时。 超声换能器的位置被跟踪使用高速摄像机以为降着陆序列。的转子和压脉袋（绿色虚线之间所作的线段之间的角度编线段）和大腿中部和压脉袋（紫色虚线段）之间的线段计算。共有16个试验，每个学科2个试验（试验1和2与受到1等）在图4中观察到。相对于解剖标志着陆过程中换能器支架的最小角度变化（0.91°±0.54°），观察在所有16项试验。超声换能器的角度变化呈高重复性以及（ICC 2,1 = 0.90，P &lt;0.05），这表明着陆试验过程中换能器的运动是最小和速度的测量并没有因任何传感器运动的影响。 二。三维运动相机及测力板我们主要侧重于膝关节和髋关节屈曲角度，膝关节外翻角和膝外翻的时刻。我们发现，与地面的初始接触期间，受试者有下列运动型态:髋fLEXION 41°±13°，膝关节屈曲23°±9°，膝外翻0.03°±6度。当他们在着陆阶段的进展，取得的最大角度为:髋关节屈曲58°±19度，膝关节屈曲54°±24°，膝外翻-4°±8度（ 图5）。膝外翻的时刻呈现减少从0.03±0.03到0.1±0.1标准立方米/公里从初始接触地面到其最大在着陆阶段（ 图6）。 <img alt="图1" src="/files/ftp_upload/50595/50595fig1.jpg" /> 图1。 腹直肌的VTDI速度测量的代表性股二头肌肌肉。灰色光束代表两个单独的发射和接收波束，红色线表示的横向速度分量（沿膝盖的近端-远端方向）和蓝色线表示轴向速度组分（沿着肌肉的厚度）。 &lt;P类="j​​ove_content"&gt; <img alt="图2" src="/files/ftp_upload/50595/50595fig2.jpg" /> 图2。在跌落着陆轴向和侧向速度进行比较，以视频帧的序列（上图）。下面板是在轴向和横向运动速度，其中A对应于初始膝屈曲，B对应于膝伸展，C对应的脚趾敲击地面，D对应于脚跟撞击地面，E对应于膝关节屈曲post登陆和F对应于伸膝和稳定。 <img alt="图3" src="/files/ftp_upload/50595/50595fig3.jpg" /> 图3。所有8个科目（每科2项试验）产生的速度矢量的幅值可重复性。男人都用红色表示的钻石和妇女在蓝色圆圈。 <img alt="图4" src="/files/ftp_upload/50595/50595fig4.jpg" /> 图4。图A中的错误线段由超声换能器保持器制成，并在大腿中部（紫色虚线段）的标记物和由超声换能器取得的线段并在转子的标记之间的角度（绿色虚线段）。面板B.在大腿中部和由超声换能器和在转子的标记所作的线段由超声换能器保持器和所述标记的线段之间的角度的绝对误差。 <img alt="图5" fo:content-width="6in" src="/files/ftp_upload/50595/50595fig5.jpg" /> 图5。图显示了降着陆任务在3D动作捕捉。 A对应于初始膝关节屈曲用于从平台上推出，B对应于脚趾撞击地面，C对应于脚跟撞击地面，D对应于膝关节屈曲后的着陆和E对应于连护膝护肘e扩展和稳定。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/50595/50595fig5hires.jpg" target="_blank">点击此处查看大图。</a> <img alt="图6" fo:content-width="6in" src="/files/ftp_upload/50595/50595fig6.jpg" /> 图6。图为代表膝外翻时刻变化过程中的下拉式跳跃的立场相。膝外翻瞬间呈现同比增长0.03±0.03至0.1±0.1牛米/从最初接触地面，其最大在着陆阶段公里。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/50595/50595fig6hires.jpg" target="_blank">点击这里查看大图。</a>

Watch this Scientific Journal Video about A Novel Application of Musculoskeletal Ultrasound Imaging at JoVE.com

A Novel Application of Musculoskeletal Ultrasound Imaging

我们描述了一个新的基于超声波的载体组织多普勒成像技术测量肌肉收缩速度，应变和应变率在动态活动亚毫秒级的时间分辨率。这种方法提供的动态肌肉功能互补的测量，并可能导致更好的了解相关肌肉骨骼疾病的机制。

肌肉骨骼超声成像的一种新应用

Ultrasound is an attractive modality for imaging muscle and tendon  motion during dynamic tasks and can provide a complementary methodological  approach ...

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Universidad de Cartagena UDEC

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JoVE Journal

Medicine

23.3K 次观看.  George Mason University. 我们描述了一个新的基于超声波的载体组织多普勒成像技术测量肌肉收缩速度，应变和应变率在动态活动亚毫秒级的时间分辨率。这种方法提供的动态肌肉功能互补的测量，并可能导致更好的了解相关肌肉骨骼疾病的机制。 

视频: A Novel Application of Musculoskeletal Ultrasound Imaging

Murine Echocardiography and Ultrasound Imaging

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1 Echocardiography provides a powerful, non-invasive tool to serially assess cardiac morphometry and function in a living animal.2 However, using this technique on mice poses unique challenges owing to the small size and rapid heart rate of these animals.3 Until recently, few ultrasound systems were capable of performing quality echocardiography on mice, and those generally lacked the image resolution and frame rate necessary to obtain truly quantitative measurements. Newly released systems such as the VisualSonics Vevo2100 provide new tools for researchers to carefully and non-invasively investigate cardiac function in mice. This system generates high resolution images and provides analysis capabilities similar to those used with human patients. Although color Doppler has been available for over 30 years in humans, this valuable technology has only recently been possible in rodent ultrasound.4,5 Color Doppler has broad applications for echocardiography, including the ability to quickly assess flow directionality in vessels and through valves, and to rapidly identify valve regurgitation. Strain analysis is a critical advance that is utilized to quantitatively measure regional myocardial function.6 This technique has the potential to detect changes in pathology, or resolution of pathology, earlier than conventional techniques. Coupled with the addition of three-dimensional image reconstruction, volumetric assessment of whole-organs is possible, including visualization and assessment of cardiac and vascular structures. Murine-compatible contrast imaging can also allow for volumetric measurements and tissue perfusion assessment.

This video demonstrates use of a rail-mounted high-frequency ultrasound probe to perform echocardiography on an anesthetized mouse. The methods describe both conventional two-dimensional and M-mode measurements of cardiac function in addition to newer, more powerful tools such as color Doppler, strain analysis, as well as general and targeted contrast imaging.

小鼠超声心动图和超声成像

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1  ...

科研

医学

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&beta; using a Whole Blood Stimulation Assay

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues as well as altered cytokine homeostasis. The innate immune system plays a crucial role in recognizing pathogens and other endogenous danger stimuli. One of the major cytokines released by innate immune cells is Interleukin (IL)-1. Therefore, we utilize a whole blood stimulation assay in order to measure the secretion of inflammatory cytokines and specifically of the pro-inflammatory cytokine IL-1&beta; 1, 2, 3.
Patients with genetic dysfunctions of the innate immune system causing autoinflammatory syndromes show an exaggerated release of mature IL-1&beta; upon stimulation with LPS alone. In order to evaluate the innate immune component of patients who present with inflammatory-associated pathologies, we use a specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns (PAMPs), such as the gram-negative bacterial endotoxin, lipopolysaccharide (LPS). These PAMPs are recognized by pathogen recognition receptors (PRRs), which are found on the cells of the innate immune system 4, 5, 6, 7. A primary signal, LPS, in conjunction with a secondary signal, ATP, is necessary for the activation of the inflammasome, a multiprotein complex that processes pro-IL-1&beta; to its mature, bioactive form 4, 5, 6, 8, 9, 10.
The whole blood assay requires minimal sample manipulation to assess cytokine production when compared to other methods that require labor intensive isolation and culturing of specific cell populations. This method differs from other whole blood stimulation assays; rather than diluting samples with a ratio of RPMI media, we perform a white blood cell count directly from diluted whole blood and therefore, stimulate a known number of white blood cells in culture 2. The results of this particular whole blood assay demonstrate a novel technique useful in elucidating patient cohorts presenting with autoinflammatory pathophysiologies.

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&#946; using a Whole Blood Stimulation Assay

We describe a simple immunoassay to measure the production of pro-inflammatory cytokines, such as IL-1 beta production, in patients presenting with autoinflammatory phenotypes. By activating cells in whole blood cultures with pathogen-associated molecular patterns, specifically with lipopolysaccharide, cytokine secretion can be conveniently evaluated in whole blood supernatants.

使用全血刺激试验的促炎性细胞因子IL -1β的精确和简单的测量

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues ...

Utilizing Transcranial Magnetic Stimulation to Study the Human Neuromuscular System

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade. While the use of TMS has expanded to the study of many systems and processes during this time, the original application and perhaps one of the most common uses of TMS involves studying the physiology, plasticity and function of the human neuromuscular system. Single pulse TMS applied to the motor cortex excites pyramidal neurons transsynaptically 2 (Figure 1) and results in a measurable electromyographic response that can be used to study and evaluate the integrity and excitability of the corticospinal tract in humans 3. Additionally, recent advances in magnetic stimulation now allows for partitioning of cortical versus spinal excitability 4,5. For example, paired-pulse TMS can be used to assess intracortical facilitatory and inhibitory properties by combining a conditioning stimulus and a test stimulus at different interstimulus intervals 3,4,6-8. In this video article we will demonstrate the methodological and technical aspects of these techniques. Specifically, we will demonstrate single-pulse and paired-pulse TMS techniques as applied to the flexor carpi radialis (FCR) muscle as well as the erector spinae (ES) musculature. Our laboratory studies the FCR muscle as it is of interest to our research on the effects of wrist-hand cast immobilization on reduced muscle performance6,9, and we study the ES muscles due to these muscles clinical relevance as it relates to low back pain8. With this stated, we should note that TMS has been used to study many muscles of the hand, arm and legs, and should iterate that our demonstrations in the FCR and ES muscle groups are only selected examples of TMS being used to study the human neuromuscular system.

Transcranial magnetic stimulation (TMS) is a non-invasive tool to gain insight on the physiology and function of the human nervous system. Here, we present our TMS techniques to study cortical excitability of the upper limb and lumbar musculature.

利用经颅磁刺激，以研究人类的神经肌肉系统

Transcranial magnetic stimulation (TMS) has been in use for more than 20 years 1, and has grown exponentially in popularity over the past decade.  While ...

MRI-guided Disruption of the Blood-brain Barrier using Transcranial Focused Ultrasound in a Rat Model

Focused ultrasound (FUS) disruption of the blood-brain barrier (BBB) is an increasingly investigated technique for circumventing the BBB1-5. The BBB is a significant obstacle to pharmaceutical treatments of brain disorders as it limits the passage of molecules from the vasculature into the brain tissue to molecules less than approximately 500 Da in size6. FUS induced BBB disruption (BBBD) is temporary and reversible4 and has an advantage over chemical means of inducing BBBD by being highly localized. FUS induced BBBD provides a means for investigating the effects of a wide range of therapeutic agents on the brain, which would not otherwise be deliverable to the tissue in sufficient concentration. While a wide range of ultrasound parameters have proven successful at disrupting the BBB2,5,7, there are several critical steps in the experimental procedure to ensure successful disruption with accurate targeting. This protocol outlines how to achieve MRI-guided FUS induced BBBD in a rat model, with a focus on the critical animal preparation and microbubble handling steps of the experiment.

Microbubble-mediated focused ultrasound disruption of the blood-brain barrier (BBB) is a promising technique for non-invasive targeted drug delivery in the brain1-3. This protocol outlines the experimental procedure for MRI-guided transcranial BBB disruption in a rat model.

MRI引导下的破坏血脑屏障在大鼠模型经颅聚焦超声

Focused ultrasound (FUS) disruption of the blood-brain barrier (BBB) is an increasingly investigated technique for circumventing the BBB1-5. The BBB is a ...

The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel underlying neurobiological mechanisms behind drug action and neurotransmitter-related disorders, such as depression and ADHD. Like most of the imaging methods (PET, SPECT, CT) it represents a progress in the investigation of brain disorders and the related function of neurotransmitter pathways in a non-invasive way with respect of the overall neuronal connectivity. Moreover it also provides the ideal tool for translation to clinical investigations. MRI, while still behind in molecular imaging strategies compared to PET and SPECT, has the great advantage to have a high spatial resolution and no need for the injection of a contrast-agent or radio-labeled molecules, thereby avoiding the repetitive exposure to ionizing radiations. Functional MRI (fMRI) is extensively used in research and clinical setting, where it is generally combined with a psycho-motor task. phMRI is an adaptation of fMRI enabling the investigation of a specific neurotransmitter system, such as serotonin (5-HT), under physiological or pathological conditions following activation via administration of a specific challenging drug.
The aim of the method described here is to assess brain 5-HT function in free-breathing animals. By challenging the 5-HT system while simultaneously acquiring functional MR images over time, the response of the brain to this challenge can be visualized. Several studies in animals have already demonstrated that drug-induced increases in extracellular levels of e.g. 5-HT (releasing agents, selective re-uptake blockers, etc) evoke region-specific changes in blood oxygenation level dependent (BOLD) MRI signals (signal due to a change of the oxygenated/deoxygenated hemoglobin levels occurring during brain activation through an increase of the blood supply to supply the oxygen and glucose to the demanding neurons) providing an index of neurotransmitter function. It has also been shown that these effects can be reversed by treatments that decrease 5-HT availability16,13,18,7. In adult rats, BOLD signal changes following acute SSRI administration have been described in several 5-HT related brain regions, i.e. cortical areas, hippocampus, hypothalamus and thalamus9,16,15. Stimulation of the 5-HT system and its response to this challenge can be thus used as a measure of its function in both animals and humans2,11.

The goal of this technique is to assess serotonin (5-HT) neurotransmitter function in the live and free-breathing animal with pharmacological magnetic resonance imaging (phMRI) and an intravenous challenge with a selective serotonin reuptake inhibitor (SSRI), fluoxetine.

挑战的药理功能磁共振成像:在临床前研究使用的5  - 羟色胺系统中的应用

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel ...

Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive surgical procedures. Although this is an effective method of assessing in vitro diaphragm activity, it involves non-survival surgery. The application of non-invasive ultrasound imaging as an in vivo procedure is beneficial since it not only reduces the number of animals sacrificed, but is also suitable for monitoring disease progression in live mice. Thus, our ultrasound imaging method may likely assist in the development of novel therapies that alleviate muscle injury induced by various respiratory diseases. Particularly, in clinical diagnoses of obstructive lung diseases, ultrasound imaging has the potential to be used in conjunction with other standard tests to detect the early onset of diaphragm muscle fatigue. In the current protocol, we describe how to accurately evaluate diaphragm contractility in a mouse model using a diagnostic ultrasound imaging technique.

Diagnostic ultrasound imaging has proven to be effective in diagnosing various respiratory diseases in human and animal subjects. We demonstrate a comprehensive ultrasound protocol utilized by Dr. Zuo's lab to analyze diaphragm kinetics specifically in mouse models. This is also a non-invasive research technique which can provide quantitative information on mouse respiratory muscle function.

小鼠膈肌功能诊断超声成像

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive ...

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone resorption. Osteoclasts can be generated in vitro from monocyte/macrophage precursor cells in the presence of certain cytokines, which promote survival and differentiation. Here, both in vivo and in vitro techniques are demonstrated, which allow scientists to study different cytokine contributions towards osteoclast differentiation, signaling, and activation. The minicircle DNA delivery gene transfer system provides an alternative method to establish an osteoporosis-related model is particularly useful to study the efficacy of various pharmacological inhibitors in vivo. Similarly, in vitro culturing protocols for producing osteoclasts from human precursor cells in the presence of specific cytokines enables scientists to study osteoclastogenesis in human cells for translational applications. Combined, these techniques have the potential to accelerate drug discovery efforts for osteoclast-specific targeted therapeutics, which may benefit millions of osteoporosis and arthritis patients worldwide.

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation of precursor cells into osteoclasts is regulated by cytokines and growth factors. Here, a novel gene transfer technique for differentiation of osteoclasts in vivo and cell culture protocols for differentiating precursor cells into osteoclasts in vitro as a method to study the effects of cytokines on osteoclastogenesis are described.

一种新型的<em&gt;在体内</em&gt;基因转移技术和<em&gt;在体外</em&gt;细胞分析为肌肉骨骼疾病骨质流失的研究

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone ...

State of the Art Cranial Ultrasound Imaging in Neonates

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a patient is unstable. In addition it is radiation-free and allows serial imaging. CUS possibilities have steadily expanded. However, in many neonatal intensive care units, these possibilities are not optimally used. We present a comprehensive approach for neonatal CUS, focusing on optimal settings, different probes, multiple acoustic windows and Doppler techniques. This approach is suited for both routine clinical practice and research purposes. In a live demonstration, we show how this technique is performed in the neonatal intensive care unit. Using optimal settings and probes allows for better imaging quality and improves the diagnostic value of CUS in experienced hands. Traditionally, images are obtained through the anterior fontanel. Use of supplemental acoustic windows (lambdoid, mastoid, and lateral fontanels) improves detection of brain injury. Adding Doppler studies allows screening of patency of large intracranial arteries and veins. Flow velocities and indices can be obtained. Doppler CUS offers the possibility of detecting cerebral sinovenous thrombosis at an early stage, creating a window for therapeutic intervention prior to thrombosis-induced tissue damage. Equipment, data storage and safety aspects are also addressed.

Cranial ultrasound (CUS) is a valuable tool for brain imaging in critically ill neonates. This video shows a comprehensive approach for neonatal (Doppler) CUS for both clinical and research purposes, including a bedside demonstration of the technique.

艺术颅超声成像在新生儿的状态

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a ...

High-frequency Ultrasound Imaging of Mouse Cervical Lymph Nodes

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal systems. HFUS has the ability to detect structures as small as 30 &#181;m, a property that has been utilized for visualizing superficial lymph nodes in rodents in brightness (B)-mode. Combining power Doppler with B-mode imaging allows for measuring circulatory blood flow within lymph nodes and other organs. While HFUS has been utilized for lymph node imaging in a number of mouse&#160; model systems, a detailed protocol describing HFUS imaging and characterization of the cervical lymph nodes in mice has not been reported. Here, we show that HFUS can be adapted to detect and characterize cervical lymph nodes in mice. Combined B-mode and power Doppler imaging can be used to detect increases in blood flow in immunologically-enlarged cervical nodes. We also describe the use of B-mode imaging to conduct fine needle biopsies of cervical lymph nodes to retrieve lymph tissue for histological&#160; analysis. Finally, software-aided steps are described to calculate changes in lymph node volume and to visualize changes in lymph node morphology following image reconstruction. The ability to visually monitor changes in cervical lymph node biology over time provides a simple and powerful technique for the non-invasive monitoring of cervical lymph node alterations in preclinical mouse models of oral cavity disease.

This protocol describes the application of high-frequency ultrasound (HFUS) for imaging mouse cervical lymph nodes. This technique optimizes visualization and quantification of cervical lymph node morphology, volume and blood flow. Image-guided biopsy of cervical lymph nodes and processing of lymph tissue for histological evaluation is also demonstrated.

鼠标颈淋巴结高频超声成像

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal ...

3D Ultrasound Imaging: Fast and Cost-effective Morphometry of Musculoskeletal Tissue

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS images are constructed from calibrated freehand 2D B-mode ultrasound images, which are positioned into a voxel array. Ultrasound (US) imaging allows quantification of muscle size, fascicle length, and angle of pennation. These morphological variables are important determinants of muscle force and length range of force exertion. The presented protocol describes an approach to determine volume and fascicle length of m.&#160;vastus lateralis and m.&#160;gastrocnemius medialis. 3DUS facilitates standardization using 3D anatomical references. This approach provides a fast and cost-effective approach for quantifying 3D morphology in skeletal muscles. In healthcare and sports, information on the morphometry of muscles is very valuable in diagnostics and/or follow-up evaluations after treatment or training.

3D ultrasound imaging (3DUS) allows fast and cost-effective morphometry of musculoskeletal tissues. We present a protocol to measure muscle volume and fascicle length using 3DUS.

3D 超声成像: 快速、经济高效的肌骨骼组织形态分析

The developmental goal of 3D ultrasound imaging (3DUS) is to engineer a modality to perform 3D morphological ultrasound analysis of human muscles. 3DUS ...