这项工作得到了全国眼科研究所资助1 R15的EY13649 - 01，个人从自然科学和加拿大工程研究理事会（NSERC）发现格兰特。特别感谢德达吉莱斯皮为金属加工，设备的设计和施工咨询。

简介自动化被动手指触觉刺激器（TAPS）是一个计算机控制系统，对皮肤的压力机结构表面，以衡量一个人的触觉空间视力。我们在杜肯大学的设备设计和建造，并修改其目前的形式在加拿大麦克马斯特大学。在这里，我们提供了一个水龙头的功能的概述。然后，我们描述设备的组成部分，由其他研究人员，以促进其繁殖。最后，我们展示水龙头如何进行心理物理学实验。 1。设备概述丝锥采用重力按压刺激表面皮肤（图1）。主体的扶手舒适俯卧在桌面上。指尖（或其他面积的皮肤进行测试）在于对表中的一个隧道。下面的表中，步进电机旋转的磁盘，最多可容纳至40个刺激件，定位下的隧道部分之一。重力向下的力施加在一个旋转杆的一端悬挂质量的。一个纵横延伸，从一个线性驱动器运输，防止移动杆。重力的影响下杆的支点，作为执行器电机，按下一个刺激一块向上穿过隧道，在皮肤上。简单的物理与接触皮肤刺激面上升的速度，执行机构的速度和触觉刺激的静力挂大规模的重量。作为执行器反向杆返回其起始位置，刺激片滴。因此，该装置适用于触觉刺激控制的力量，速度，接触时间，并刺激间的时间间隔。主题选择一个按钮来注册一个响应与非测试手。 TAPS的检测响应和收益提供未来的刺激。 <img src="/files/ftp_upload/1374/Fig1.png" alt="图1" /> 图1。物理学的水龙头。丝锥控制发病速度和稳定状态的刺激力。 发病速度：从线性驱动器横梁（红色）移动到右边以速度V 1，旋转杆的左端，在重力的作用下滴;杆的右端，因此对刺激件以上的印刷机手指以速度V 2 = V 1（L 2 / L 3）。因为V 1是在计算机控制下，V 2，可设置为调查所需的任何值。 力：重力施加一个向下的力，F 1 =镁的质量为M，（其中g = 9.8米/秒2）。这将导致对手指 ​​按下一个向上的力，F 2的刺激一块。对于无质量杆，F 2 = F 1（L - 1 / L，2）。由于棒是不是事实无质量，F 2是凭经验确定，由力传感器测量。研究者要进行微调到F 2，可以改变的M沿杆的位置。 F 2 M是向左移动略有增加（增加L - 1），并略有下降，M是向右移动（降低L - 1）。大众要做出较大调整到F 2，可以改变的。 2。组件和器件功能的详细信息一移动部件丝锥使用两个步进电机驱动线性驱动器运输，和其他含有刺激件旋转的磁盘。这两个马达控制丝锥的所有运动部件。这些部件趴在一个较低的表主体的扶手从上表是机械耦合。由于这两个表的耦合，电机造成的震动不会传输到皮肤。表仔细对齐，通过上表的隧道，直接旋转棒接触刺激一块以上。 我们加工的旋转盘的圆周周围40平方米插槽，以适应刺激件。我们构建了全方位的塑料棒，加工棒刺激件有适合的磁盘插槽的方形轴。这将确保件不其插槽内旋转旋转的磁盘。我们切成片圆面孔刺激的表面。 当执行器托架马达前进，对面悬挂质量的旋转杆结束上升到选定的刺激一块底部联系。由于杆描述成一个弧形向上摆动，这是一个轻巧的塑料轴承轮刺激片的底部，以消除摩擦配合。这种塑料轮子轻微转动，因为它推动了Stimulus片向上，无塑料轮，杆端略有横向拉的一块基地（对图左1），通过上表造成堵塞在隧道内的一块。为了进一步确保平稳上升轨道的一块，该隧道是适合用塑料的衣领，斜面直接向隧道中心的一块一块上升。 II。传感器 TAPS的六个传感器的使用，以确保正常运作：两个主场传感器，三个限制传感器，力传感器。 计算机驱动器和磁盘家传感器信号，执行器托架和磁盘在他们所需的初始位置是在实验的开始。执行器主页传感器是霍尔效应传感器;磁盘家传感器是一个U形的光电传感器，激活时，一块僵硬纸粘到磁盘RIM打破了红外传感器束。当磁盘在家庭的地位，是一个特定的插槽对齐下的隧道，直接在上层表。随后的所有磁盘的运动计算是相对于这个著名的起始位置。 三个限制传感器发生故障的情况下，作为安全功能。为了防止万一控制失灵的执行机构马车失控运动，驱动器的正向和反向限位传感器（霍尔效应传感器）配合。如果激活，这些执行机构切断电源。为了防止事件刺激片成为滞留在隧道中的磁盘的运动，一个红外线反射传感器连接到上层表底部的表面。一个升高到足以进入隧道的一块打破这种传感器的红外光束投，和块命令到磁盘步进电机的电压。 第六次也是最后一次的传感器，力传感器，取决于主体的指甲轻轻地（图2）。该传感器检测向下进入隧道，向前或向后手指的运动。该方案丢弃在其中的运动发生的试验。由于传感器连接到一个刚性的手臂，另外防止手指从接触后被动上升一块向上移动。由于我们使用水龙头测试被动式（手指固定）空间触觉敏锐，力传感器是一个至关重要的因素。对手指运动的最后的预防措施，我们轻轻地对手指的两侧，以防止横向运动场所塑料障碍。 <img src="/files/ftp_upload/1374/fig2a.png" alt="图2a" /> <img src="/files/ftp_upload/1374/fig2b.png" alt="图2b" /><img src="/files/ftp_upload/1374/fig2c.png" alt="图2c" /> 图2。B.力传感器来检测手指的运动。A.力传感器在90度印刷机附近指甲表面的角质层，50和80克之间的力量，榜样的力量传感器跟踪显示一秒钟前接触一秒钟保持固定的手指接触，用手指刺激一块。请注意，接触（时间= 0）时，刺激件从下面的力量推动的手指更加坚定了对传感器（有效跟踪斜坡上升）。C.如果试图向下移动手指的主题，以更好地感受到刺激表面，然后指甲和传感器之间的力量减弱（箭头）。该传感器还寄存器力波动，如果这个问题向前或向后移动手指。水龙头编程放弃任何试验，其中超过一个门槛金额（例如，20克）的传感器和指甲波动的力量之间的接触，而皮肤与刺激表面接触。使用计算机生成的声音，水龙头，可以选择性地警告检测运动的主题。 III。振动和声音阻尼 TAPS的“步进马达产生振动，而这又反过来导致声音。振动和声音都是不可取的。振动，如果传送到皮肤上，可能会干扰主体的能力来执行的触觉任务。随着时间的推移，振动也可能导致在丝锥的组件的机械不稳定。声音可能会分散主题。基于这些原因，我们已采取措施，以减少震动和声音，并达到主体，以防止任何震动。 最重要的是，下限和上限的表不互相联络。只有在这个意义上，它们是连接，它们都接触地板。因此，振动电机，较低的表的其余部分，不传送到主体的手臂，在于上层表。此外，减少震动和声音如下：较低的表玻璃纤维窗帘封闭围巾设备声音; PVC / FIBER钢筋夹着之间的执行机构，其基地的执行机构产生的振动吸收一些抗振动垫;执行器基地坐落在四个橡胶圆柱的夹心坐骑，这有助于进一步抑制振动;磁盘步进电机基地也坐在橡胶坐骑（注射器柱塞橡胶）和硅胶O型圈环绕的刺激件轴，减少磁盘转动的哒。 IV。刺激饮片丝锥使用修改后的直径为半英寸的棒，按空间结构的表面，对皮肤（图3）。棒加工有方形的竖井。棒轮面临含有刺激面;棒的方轴放入旋转盘的圆周周围40平方米插槽，确保刺激表面保持其应有的磁盘旋转方向。 任何刺激的表面可能被加工成棒的面孔。我们用的水龙头主要适用于方波光栅（即平行脊和沟槽）。要创建这些，我们开始用直径0.5“聚甲醛塑料圆柱棒，长3”。我们使用Sherline铣床，切割棒2.75“，2”长正方形截面轴（8.7 × 8.7毫米），和一个0.75“长，12.74毫米直径的圆形圆柱头，然后我们碾槽，在创建刺激表面的圆脸。每个刺激表面平等脊和沟槽的宽度，我们做了件槽宽度范围从0.25至3.10毫米，0.15毫米递增。我们这20槽宽度各为两个相同的刺激件，共40件。然后我们插入磁盘等，是代表在每个槽宽度平方米插槽件垂直（手指长轴平行）和水平（横向手指长轴）方向。 <img src="/files/ftp_upload/1374/Fig3.png" alt="图3" /> 图3。刺激片。每个刺激表面磨成面对一个直径为0.5英寸的聚甲醛棒。被切断，有一个方形截面杆的基地，允许它在旋转盘的方形切口放入。显示片的表面是一个方波光栅，与平行的槽脊的宽度相等。 五，计算机控制水龙头一个完全自动化，计算机控制系统。许多计算机硬件和软件配置，可用于控制设备。我们目前使用的是Macintosh G3运行的LabVIEW 6.1（美国国家仪器）。计算机通信通过一个步进电机驱动器，命令驱动器和磁盘电机，并阅读的家庭传感器和限位开关的电机控制器板。一个PCI数据采集板读取力传感器和受响应按钮。 VI。部队和时序限制力范围：我们用水龙头提供从10到50克不等的力量刺激。我们怀疑，在组件的摩擦会妨碍交付的刺激下约5克的力量，和一些机械不稳定，会出现超过100克左右的力量。 刺激间的时间间隔（ISI）：刺激的表面和手指之间的距离，以及线性驱动器和磁盘步进电机的稳定速度，以及设置该设备可以实现对ISI的下限。我们估计在1.5秒左右，这个限制。因此，该设备是不适合作为掩蔽实验，要求亚秒级的三军情报局等应用。 3。进行实验在这里，我们描述了一个协议，可以使用的水龙头：两间隔被迫的选择（2国际金融公司）光栅方向任务（GOT）与贝叶斯自适应跟踪。 TAPS可以编程，以及许多其他心理物理学协议。我们已经用它先前进行跟踪与楼梯1,2和2国际金融公司的实验，不断刺激的方法。当然，它是能够进行是/否程序，以及2国际金融公司。 2，国际金融公司了任务，对于我们使用方波光栅接触用手指远端垫4厘米/ GM的接触力，50秒的发病速度，1秒的接触时间。每个试验包括两个连续的刺激简报（ISI：2秒）与沟槽宽度相同的光栅，但不同的方向为90度。在一个演示文稿，槽垂直对齐（平行），并在其他水平（横向），手指的长轴。刺激的顺序是由计算机程序随机选择。主题表示水平方向是否发生在第一或第二的时间间隔，与非测试手按两个按钮之一。一个贝叶斯自适应方法调整克roove宽度从审判审判。 我们编程丝锥与修改后的版本的PSI（Ψ）的方法，贝叶斯自适应算法3。这种方法有几个优点，比传统的自适应方法，如楼梯方法，。首先，PSI的方法，估计不只是一个单点上的心理功能（例如，71％的门槛，估计由2跌1的楼梯），但整个的心理功能。其次，该方法产生心理功能参数的贝叶斯后验概率分布（从而置信区间）。最后，该方法是有效的。它拥有在可能的心理功能的形状记忆数以千计，并在更新后，每个响应每个函数的后验概率。然后，它使用预期的熵最小化程序，选择下一个刺激，这就是，它选择对每个试验，预计将最大限度地发挥信息增益的刺激。 Kontsevich和泰勒3之后，我们仿照作为一个刺激水平的电源功能ð -首相，每个主题的心理功能- P彗星（X），一个刺激水平，x的函数正确的反应的可能性-的混合物一个累积的正常（概率）功能和递减率长期： <img src="/files/ftp_upload/1374/eq1.png" alt="公式1" /> <img src="/files/ftp_upload/1374/eq2.png" alt="公式（2）" /> 在这里， 一个是阈值（对应76％的正确反应概率的刺激水平，D -首相= 1），b是斜率为主体的心理功能，。 我们修改当作一个未知的值的参数的递减率（增量）Ψ算法，并通过计算每次试验后的“猜测贝叶斯因子”： <img src="/files/ftp_upload/1374/eq3.png" alt="公式3" /> 这贝叶斯因子是一种似然比，根据假设1中的数据进行比较的可能性，主体是猜测，根据假设2数据的概率，这一主题有一个心理功能。分子为主体的数据的概率，D组（在每个凹槽宽度的正确和不正确的反应）的主题仅仅是猜测在所有的试验，包括目前的审判（50％的概率正确的）。分母是算法的主体的心理功能的最佳估计数据的可能性。根据我们的经验，最年轻的学科的贝叶斯因子迅速接近零测试块的进展，表明主体的表现，符合一个心理功能。对于一些年长者，贝叶斯因子高于1，表明这个问题是无法执行任务。 TAPS可以编程终止测试块，如果猜测贝叶斯因素已经超过阈值后指定数量的试验。 4。结果图4显示了实验的块组成的40次试验，从2国际金融公司GOT的实验。这40试块由80刺激主体的右手中指，1秒钟的接触时间和2秒的三军情报局之间在一个单一的审判接触。主体的76％正确的阈值是1.7毫米，后验概率密度函数（PDF）的阈值参数（C组）的模式。为主体的阈值参数后的PDF的宽度表示，95％可信区间为1.3 - 1.9毫米。 <img src="/files/ftp_upload/1374/fig4a.png" alt="图4a" /><img src="/files/ftp_upload/1374/fig4b.png" alt="图4b" /> <img src="/files/ftp_upload/1374/fig4c.png" alt="图4c" /> 图4 2间隔被迫选择光栅定位任务的结果。A.一个实验块，在这种情况下适用于右手中指的40试验。这个题目的正确（加号）和在不同的凹槽宽度不正确（空心圆）的反应是策划对试验数。一个贝叶斯自适应方法（见文）确定的凹槽宽度适用于基于主体的表现，序列。B.适合这个问题的最好的心理功能。C.后 ​​验概率密度函数为主体的76％正确的槽宽（心理功能的一个参数，对应于D -首相= 1）。

<ol>
	<li>Goldreich, D. &. a. m. p. ;. a. m. p., Kanics, I. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tactile+acuity+is+enhanced+in+blindness.">Tactile acuity is enhanced in blindness.</a> J. Neurosci. 23, 3439-3445 (2003).</li><li>Goldreich, D. &. a. m. p. ;. a. m. p., Kanics, I. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performance+of+blind+and+sighted+humans+on+a+tactile+grating+detection+task.">Performance of blind and sighted humans on a tactile grating detection task.</a> Percept. Psychophys. 68, 1363-1371 (2006).</li><li>Kontsevich, L. L. &. a. m. p. ;. a. m. p., Tyler, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bayesian+adaptive+estimation+of+psychometric+slope+and+threshold.">Bayesian adaptive estimation of psychometric slope and threshold.</a> Vision. Res. 39, 2729-2737 (1999).</li><li>Bleyenheuft, Y., Cols, C., Arnould, C. &. a. m. p. ;. a. m. p., Thonnard, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-related+changes+in+tactile+spatial+resolution+from+6+to+16+years.">Age-related changes in tactile spatial resolution from 6 to 16 years.</a> Somatosens. Mot. Res. 23, 83-87 (2006).</li><li>Johnson, K. O. &. a. m. p. ;. a. m. p., Phillips, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tactile+spatial+resolution.+I.+Two-point+discrimination,+gap+detection,+grating+resolution,+and+letter+recognition.">Tactile spatial resolution. I. Two-point discrimination, gap detection, grating resolution, and letter recognition.</a> J. Neurophysiol. 46, 1177-1192 (1981).</li><li>Gibson, G. O. &. a. m. p. ;. a. m. p., Craig, 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=The+effect+of+force+and+conformance+on+tactile+intensive+and+spatial+sensitivity.">The effect of force and conformance on tactile intensive and spatial sensitivity.</a> Exp. Brain Res. 170, 172-181 (2006).</li><li>Johnson, K. O. &. a. m. p. ;. a. m. p., Phillips, J. 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+rotating+drum+stimulator+for+scanning+embossed+patterns+and+textures+across+the+skin.">A rotating drum stimulator for scanning embossed patterns and textures across the skin.</a> J. Neurosci. Methods. 22, 221-231 (1988).</li><li>Craig, 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=Grating+orientation+as+a+measure+of+tactile+spatial+acuity.">Grating orientation as a measure of tactile spatial acuity.</a> Somatosens. Mot. Res. 16, 197-206 (1999).</li><li>Killebrew, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+dense+array+stimulator+to+generate+arbitrary+spatio-temporal+tactile+stimuli.">A dense array stimulator to generate arbitrary spatio-temporal tactile stimuli.</a> J. Neurosci. Methods. 161, 62-74 (2007).</li></ol>

空间结构的机械性刺激的控制下交付对面临的挑战不是在视觉或听觉刺激，可以使用市售的设备（电脑屏幕，音响喇叭）的交付。出于这个原因，许多触觉心理物理学实验仍然使用手册刺激交付。 触觉刺激的手动交付是费时，而且需要极大的关怀和对调查的一部分浓度。例如，Bleyenheuft等4报告说，“大约1-2毫米的皮肤垂直变形的手动应用... ...考官特别注意，以避免任何可能扭曲措施之间的皮肤和光栅的剪应力。 ... ...“也许是因为人工刺激交付相对缓慢和集中密集，使用这种方法的许多研究都受雇于是/否（只是每一个试验的刺激），而不是两个区间被迫选择协议。 不幸的是，即使采取非常谨慎的，人工刺激交付留下许多的刺激参数失控。这些参数包括刺激的力量，发病速度，持续时间，和对皮肤的刺激表面的稳定。这些参数的，也许是最好的研究及其对性能的影响是刺激力。人类在光栅上的指尖从500到1200微米5压痕深度变化的倾向歧视逐步改善;性能比50克力 1，10克力之间的50克和200 克 6的观察虽然没有进一步改善。对于其他应用，如光栅检测（区分从沟槽表面的光滑），有效的增加显着提高整个10至200克2,6范围内的性能。 为了克服这些挑战与手工测试相关，我们开发的自动化系统，水龙头。我们在建设水龙头的目的是创建一个简单，安全，多功能，高效和负担得起的的设备控制的触觉刺激的应用。重力控制的使用是一种简单，安全的办法适用于触觉刺激。安全保证，因为对皮肤的力量不能超过，由于重力的作用。强大的线性驱动器，转移到皮肤上的重力，然后撤回接触。该设备是通用的，因为它会容纳任何刺激表面到半英寸直径的棒的两端（或附加上）加工，并可以通过编程来运行各种心理物理学的协议。水龙头高效，迅速部署的刺激，可以运行省时的自适应算法。最后，水龙头组件小额赠款持有人负担得起。电机成本约为1,500元;（不包括计算机，但包括PCI板和步进电机驱动器），电子元件，约6000美元。约$ 1,000可购买一个微型铣床及配件，或专业的机械师可以聘请到轧机的刺激件和其他小零件。 我们希望这个系统将被他人复制，并有助于促进对触觉控制的测试，这在近年来已经看到了可喜的进展 5,7,8,9移动。

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Delrin plastic rods 0.5&#x201D; diameter 3&#x201D; long </td>
<td>&nbsp;</td>
<td>Small Parts</td>
<td>ZRD-08</td>
<td>Rods from which the stimulus pieces are made</td>
</tr>
<tr>
<td>Chrome-steel bearing, 0.5" bore diameter</td>
<td>&nbsp;</td>
<td>Small Parts</td>
<td>BR-08-01</td>
<td>Pivot for the rotating rod</td>
</tr>
<tr>
<td>Plastic bearing, 0.5&#x201D; bore, 1.125&#x201D; outer diameter, 0.25&#x201D; thick</td>
<td>&nbsp;</td>
<td>Small Parts</td>
<td>BRP-08-01</td>
<td>Wheel at end of rotating rod</td>
</tr>
<tr>
<td>Fiberglass curtain, 0.75&#x201D; thick</td>
<td>&nbsp;</td>
<td>McMaster-Carr</td>
<td>9781T83</td>
<td>Sound absorbing curtain around lower table</td>
</tr>
<tr>
<td>Silicone O-rings 5/16" inside diameter</td>
<td>&nbsp;</td>
<td>McMaster-Carr</td>
<td>AS568A- 011</td>
<td>To reduce vibration of stimulus pieces</td>
</tr>
<tr>
<td>Anti-Vibration Pads 2"x 2", 93 PSI max </td>
<td>&nbsp;</td>
<td>McMaster-Carr</td>
<td>60105K61</td>
<td>Put under the linear actuator</td>
</tr>
<tr>
<td>Rubber sandwich mounts 1/2" H x 3/8" W </td>
<td>&nbsp;</td>
<td>McMaster-Carr</td>
<td>9378K11</td>
<td>Put under linear actuator base</td>
</tr>
<tr>
<td>Stepper motor Nema 23, 3-stack </td>
<td>&nbsp;</td>
<td>Industrial Devices</td>
<td>S23</td>
<td>For rotating the stimulus disk</td>
</tr>
<tr>
<td>Linear rodless actuator</td>
<td>&nbsp;</td>
<td>Industrial Devices</td>
<td>R2S23N-105A-18-l-M35M</td>
<td>To control rotating bar movement</td>
</tr>
<tr>
<td>1 Hall effect switch &#x201C;normally open&#x201D; config. </td>
<td>&nbsp;</td>
<td>Industrial Devices</td>
<td>RP1</td>
<td>Actuator home sensor</td>
</tr>
<tr>
<td>2 Hall effect switches &#x201C;normally closed&#x201D; config. </td>
<td>&nbsp;</td>
<td>Industrial Devices</td>
<td>RP2</td>
<td>Actuator limit sensors</td>
</tr>
<tr>
<td>Micro switch</td>
<td>&nbsp;</td>
<td>Honeywell</td>
<td>FSG15N1A</td>
<td>Finger force sensor</td>
</tr>
<tr>
<td>Retro-reflective sensor</td>
<td>&nbsp;</td>
<td>Honeywell</td>
<td>FE7B-RB6VG-M</td>
<td>Piece position sensor</td>
</tr>
<tr>
<td>Motor controller board, closed-loop control</td>
<td>&nbsp;</td>
<td>National Instruments</td>
<td>PCI-step-4CX</td>
<td>Mediates communication between computer and Nudrive</td>
</tr>
<tr>
<td>Nudrive stepper motor driver</td>
<td>&nbsp;</td>
<td>National Instruments</td>
<td>Nudrive 4SX-211</td>
<td>Sends command voltages to the linear actuator and disk stepper motors</td>
</tr>
<tr>
<td>Data acquisition board</td>
<td>&nbsp;</td>
<td>National Instruments</td>
<td>PCI-MIO-16E-1</td>
<td>Reads finger force sensor</td>
</tr>
<tr>
<td>LabVIEW</td>
<td>&nbsp;</td>
<td>National Instruments</td>
<td>&nbsp;</td>
<td>Programming language</td>
</tr>
<tr>
<td>Valuemotion library</td>
<td>&nbsp;</td>
<td>National Instruments</td>
<td>&nbsp;</td>
<td>Motor control routines</td>
</tr>
<tr>
<td>Photoelectric sensor</td>
<td>&nbsp;</td>
<td>Panasonic</td>
<td>PM-K53-C1</td>
<td>Disk home sensor</td>
</tr>
<tr>
<td>Mini milling machine</td>
<td>&nbsp;</td>
<td>Sherline</td>
<td>2010-DRO</td>
<td>For machining the stimulus surfaces</td>
</tr>
<tr>
<td>Ohaus precision mass set</td>
<td>&nbsp;</td>
<td>Edmund Scientific</td>
<td>&nbsp;</td>
<td>Mass on rotating bar</td>
</tr>
<tr>
<td>Parrish Magic Line aluminum cake pan bottom, 9&#x201D; diam.</td>
<td>&nbsp;</td>
<td>Sur La Table</td>
<td>&nbsp;</td>
<td>Rotating disk machined with square cut-outs to hold the stimulus pieces</td>
</tr>		</tbody>
		</table>
		</div>

a tactile automated passive-finger stimulator (taps)

虽然触觉空间视力测试是在神经科学的研究和临床评估的存在，少数自动化设备提供控制空间结构的刺激皮肤。因此，调查通常适用于手动触觉刺激。手动刺激应用程序费时，需要极大的关怀和浓度对调查的一部分，并留下许多刺激参数失控。我们在这里描述的一个计算机控制的触觉刺激系统，自动化被动手指的触觉刺激器（TAPS），适用于空间结构对皮肤的刺激，发病速度，接触力和接触时间，控制。水龙头一个功能强大的可编程系统，能够有效地开展各种心理物理学程序。我们描述的水龙头组件，并展示如何使用水龙头管理两个区间被迫选择触觉光栅方向测试。 通讯作者：丹尼尔Goldreich

Watch this Scientific Journal Video about A Tactile Automated Passive-Finger Stimulator TAPS at JoVE.com

A Tactile Automated Passive-Finger Stimulator TAPS

我们描述了一个计算机控制的设备，为调查触觉：自动化被动手指触觉刺激器（TAPS）。我们描述的水龙头组件，并展示如何使用水龙头管理两个区间被迫选择触觉光栅方向测试。

自动化被动手指触觉刺激器（TAPS）

Although tactile spatial acuity tests are used in both neuroscience research and clinical assessment, few automated devices exist for delivering ...

a-tactile-automated-passive-finger-stimulator-taps

Research

JoVE Journal

Biology

A Tactile Automated Passive-Finger Stimulator (TAPS)

13.6K 次观看.  Duquesne University. 我们描述了一个计算机控制的设备，为调查触觉：自动化被动手指触觉刺激器（TAPS）。我们描述的水龙头组件，并展示如何使用水龙头管理两个区间被迫选择触觉光栅方向测试。

视频: A Tactile Automated Passive-Finger Stimulator TAPS

Semi-automated Optical Heartbeat Analysis of Small Hearts

We have developed a method for analyzing high speed optical recordings from Drosophila, zebrafish and embryonic mouse hearts (Fink, et. al., 2009). Our Semi-automatic Optical Heartbeat Analysis (SOHA) uses a novel movement detection algorithm that is able to detect cardiac movements associated with individual contractile and relaxation events. The program provides a host of physiologically relevant readouts including systolic and diastolic intervals, heart rate, as well as qualitative and quantitative measures of heartbeat arrhythmicity. The program also calculates heart diameter measurements during both diastole and systole from which fractional shortening and fractional area changes are calculated. Output is provided as a digital file compatible with most spreadsheet programs. Measurements are made for every heartbeat in a record increasing the statistical power of the output. We demonstrate each of the steps where user input is required and show the application of our methodology to the analysis of heart function in all three genetically tractable heart models.

We have developed a Semi-automated Optical Heartbeat Analysis method (SOHA) for analyzing high speed optical recordings from Drosophila, zebrafish and embryonic mouse hearts. We demonstrate the application of our methodology to the analysis of heart function in fruit fly and embryonic mouse hearts.

半自动小红桃光学心跳分析

We have developed a method for analyzing high speed optical recordings from Drosophila, zebrafish and embryonic mouse hearts (Fink, et. al., 2009). Our ...

科研

生物学

Digital Microfluidics for Automated Proteomic Processing

Clinical proteomics has emerged as an important new discipline, promising the discovery of biomarkers that will be useful for early diagnosis and prognosis of disease. While clinical proteomic methods vary widely, a common characteristic is the need for (i) extraction of proteins from extremely heterogeneous fluids (i.e. serum, whole blood, etc.) and (ii) extensive biochemical processing prior to analysis. Here, we report a new digital microfluidics (DMF) based method integrating several processing steps used in clinical proteomics. This includes protein extraction, resolubilization, reduction, alkylation and enzymatic digestion. Digital microfluidics is a microscale fluid-handling technique in which nanoliter-microliter sized droplets are manipulated on an open surface. Droplets are positioned on top of an array of electrodes that are coated by a dielectric layer - when an electrical potential is applied to the droplet, charges accumulate on either side of the dielectric. The charges serve as electrostatic handles that can be used to control droplet position, and by biasing a sequence of electrodes in series, droplets can be made to dispense, move, merge, mix, and split on the surface. Therefore, DMF is a natural fit for carrying rapid, sequential, multistep, miniaturized automated biochemical assays. This represents a significant advance over conventional methods (relying on manual pipetting or robots), and has the potential to be a useful new tool in clinical proteomics.
Mais J. Jebrail, Vivienne N. Luk, and Steve C. C. Shih contributed equally to this work.
Sergio L. S. Freire's current address is at the University of the Sciences in Philadelphia located at 600 South 43rd Street, Philadelphia, PA 19104.

Digital Microfluidics is a technique characterized by the manipulation of discrete droplets (~nL - mL) on an array of electrodes by the application of electrical fields. It is well-suited for carrying out rapid, sequential, miniaturized automated biochemical assays. Here, we report a platform capable of automating several proteomic processing steps.

数字微流体进行自动蛋白质组学处理

Clinical proteomics has emerged as an important new discipline, promising the discovery of biomarkers that will be useful for early diagnosis and ...

Development of automated imaging and analysis for zebrafish chemical screens. 

We demonstrate the application of image-based high-content screening (HCS) methodology to identify small molecules that can modulate the FGF/RAS/MAPK pathway in zebrafish embryos. The zebrafish embryo is an ideal system for in vivo high-content chemical screens. The 1-day old embryo is approximately 1mm in diameter and can be easily arrayed into 96-well plates, a standard format for high throughput screening. During the first day of development, embryos are transparent with most of the major organs present, thus enabling visualization of tissue formation during embryogenesis. The complete automation of zebrafish chemical screens is still a challenge, however, particularly in the development of automated image acquisition and analysis. We previously generated a transgenic reporter line that expresses green fluorescent protein (GFP) under the control of FGF activity and demonstrated their utility in chemical screens 1. To establish methodology for high throughput whole organism screens, we developed a system for automated imaging and analysis of zebrafish embryos at 24-48 hours post fertilization (hpf) in 96-well plates 2. In this video we highlight the procedures for arraying transgenic embryos into multiwell plates at 24hpf and the addition of a small molecule (BCI) that hyperactivates FGF signaling 3. The plates are incubated for 6 hours followed by the addition of tricaine to anesthetize larvae prior to automated imaging on a Molecular Devices ImageXpress Ultra laser scanning confocal HCS reader. Images are processed by Definiens Developer software using a Cognition Network Technology algorithm that we developed to detect and quantify expression of GFP in the heads of transgenic embryos. In this example we highlight the ability of the algorithm to measure dose-dependent effects of BCI on GFP reporter gene expression in treated embryos.

Development of automated imaging and analysis for zebrafish chemical screens.

We report the development of a system for automated imaging and analysis of zebrafish transgenic embryos in multiwell plates. This demonstrates the ability to measure dose dependent effects of a small molecule, BCI, on Fibroblast Growth Factor reporter gene expression and provide technology for establishing high-throughput zebrafish chemical screens.

斑马鱼化工屏幕自动成像和分析的发展。

We demonstrate the application of image-based high-content screening (HCS) methodology to identify small molecules that can modulate the FGF/RAS/MAPK ...

Using an Automated Cell Counter to Simplify Gene Expression Studies: siRNA Knockdown of IL-4 Dependent Gene Expression in Namalwa Cells

The use of siRNA mediated gene knockdown is continuing to be an important tool in studies of gene expression.  siRNA studies are being conducted not only to study the effects of downregulating single genes, but also to interrogate signaling pathways and other complex interaction networks.  These pathway analyses require both the use of relevant cellular models and methods that cause less perturbation to the cellular physiology.  Electroporation is increasingly being used as an effective way to introduce siRNA and other nucleic acids into difficult to transfect cell lines and primary cells without altering the signaling pathway under investigation. There are multiple critical steps to a successful siRNA experiment, and there are ways to simplify the work while improving the data quality at several experimental stages.  To help you get started with your siRNA mediated gene knockdown project, we will demonstrate how to perform a pathway study complete from collecting and counting the cells prior to electroporation through post transfection real-time PCR gene expression analysis.  The following study investigates the role of the transcriptional activator STAT6 in IL-4 dependent gene expression of CCL17 in a Burkitt lymphoma cell line (Namalwa).  The techniques demonstrated are useful for a wide range of siRNA-based experiments on both adherent and suspension cells.  We will also show how to streamline cell counting with the TC10 automated cell counter, how to electroporate multiple samples simultaneously using the MXcell electroporation system, and how to simultaneously assess RNA quality and quantity with the Experion automated electrophoresis system.

This procedure describes a quick and easy workflow to introduce siRNA into difficult to transfect cell lines and follow gene expression by real-time PCR. Use of an automated cell counter, multi-well electroporation plate, and automated electrophoresis station provide quick and reliable results without the need for expensive robotic handling.

使用自动细胞计数器简化基因表达研究：在Namalwa细胞的IL - 4依赖的基因表达的siRNA击倒

The use of siRNA mediated gene knockdown is continuing to be an important tool in studies of gene expression.  siRNA studies are being conducted not only ...

LeafJ: An ImageJ Plugin for Semi-automated Leaf Shape Measurement

High throughput phenotyping (phenomics) is a powerful tool for linking genes to their functions (see review1 and recent examples2-4). Leaves are the primary photosynthetic organ, and their size and shape vary developmentally and environmentally within a plant. For these reasons studies on leaf morphology require measurement of multiple parameters from numerous leaves, which is best done by semi-automated phenomics tools5,6. Canopy shade is an important environmental cue that affects plant architecture and life history; the suite of responses is collectively called the shade avoidance syndrome (SAS)7. Among SAS responses, shade induced leaf petiole elongation and changes in blade area are particularly useful as indices8. To date, leaf shape programs (e.g. SHAPE9, LAMINA10, LeafAnalyzer11, LEAFPROCESSOR12) can measure leaf outlines and categorize leaf shapes, but can not output petiole length. Lack of large-scale measurement systems of leaf petioles has inhibited phenomics approaches to SAS research. In this paper, we describe a newly developed ImageJ plugin, called LeafJ, which can rapidly measure petiole length and leaf blade parameters of the model plant Arabidopsis thaliana. For the occasional leaf that required manual correction of the petiole/leaf blade boundary we used a touch-screen tablet. Further, leaf cell shape and leaf cell numbers are important determinants of leaf size13. Separate from LeafJ we also present a protocol for using a touch-screen tablet for measuring cell shape, area, and size. Our leaf trait measurement system is not limited to shade-avoidance research and will accelerate leaf phenotyping of many mutants and screening plants by leaf phenotyping.

Demonstration of key methods for high throughput leaf measurements. These methods can be used to accelerate leaf phenotyping when studying many plant mutants or otherwise screening plants by leaf phenotype.

LeafJ:ImageJ的插件半自动叶形状测量

High throughput phenotyping (phenomics) is a powerful tool for linking genes to their functions (see review1 and recent examples2-4). Leaves are the ...

Semi-automated Imaging of Tissue-specific Fluorescence in Zebrafish Embryos

Zebrafish embryos are a powerful tool for large-scale screening of small molecules. Transgenic zebrafish that express fluorescent reporter proteins are frequently used to identify chemicals that modulate gene expression. Chemical screens that assay fluorescence in live zebrafish often rely on expensive, specialized equipment for high content screening. We describe a procedure using a standard epifluorescence microscope with a motorized stage to automatically image zebrafish embryos and detect tissue-specific fluorescence. Using transgenic zebrafish that report estrogen receptor activity via expression of GFP, we developed a semi-automated procedure to screen for estrogen receptor ligands that activate the reporter in a tissue-specific manner. In this video we describe procedures for arraying zebrafish embryos at 24-48 hours post fertilization (hpf) in a 96-well plate and adding small molecules that bind estrogen receptors. At 72-96 hpf, images of each well from the entire plate are automatically collected and manually inspected for tissue-specific fluorescence. This protocol demonstrates the ability to detect estrogens that activate receptors in heart valves but not in liver.

Described here is a protocol for semi-automated imaging of tissue-specific fluorescence in zebrafish embryos.

组织特异性荧光的斑马鱼胚胎半自动成像

Zebrafish embryos are a powerful tool for large-scale screening of small molecules. Transgenic zebrafish that express fluorescent reporter proteins are ...

Automated Analysis of Dynamic Ca2+ Signals in Image Sequences

Intracellular Ca2+ signals are commonly studied with fluorescent Ca2+ indicator dyes and microscopy techniques. However, quantitative analysis of Ca2+ imaging data is time consuming and subject to bias. Automated signal analysis algorithms based on region of interest (ROI) detection have been implemented for one-dimensional line scan measurements, but there is no current algorithm which integrates optimized identification and analysis of ROIs in two-dimensional image sequences. Here an algorithm for rapid acquisition and analysis of ROIs in image sequences is described. It utilizes ellipses fit to noise filtered signals in order to determine optimal ROI placement, and computes Ca2+ signal parameters of amplitude, duration and spatial spread. This algorithm was implemented as a freely available plugin for ImageJ (NIH) software. Together with analysis scripts written for the open source statistical processing software R, this approach provides a high-capacity pipeline for performing quick statistical analysis of experimental output. The authors suggest that use of this analysis protocol will lead to a more complete and unbiased characterization of physiologic Ca2+ signaling.

Automated Analysis of Dynamic Ca2+ Signals in Image Sequences

Here a novel region of interest analysis protocol based on sorting best-fit ellipses assigned to regions of positive signal within two-dimensional time lapse image sequences is demonstrated. This algorithm may enable investigators to comprehensively analyze physiological Ca2+ signals with minimal user input and bias.

动态钙的自动分析<sup&gt; 2 +</sup&gt;在图像序列信号

Intracellular Ca2+ signals are commonly studied with fluorescent Ca2+ indicator dyes and microscopy techniques. However, quantitative analysis of Ca2+ ...

High-resolution Time-lapse Imaging and Automated Analysis of Microtubule Dynamics in Living Human Umbilical Vein Endothelial Cells

The physiological process by which new vasculature forms from existing vasculature requires specific signaling events that trigger morphological changes within individual endothelial cells (ECs). These processes are critical for homeostatic maintenance such as wound healing, and are also crucial in promoting tumor growth and metastasis. EC morphology is defined by the organization of the cytoskeleton, a tightly regulated system of actin and microtubule (MT) dynamics that is known to control EC branching, polarity and directional migration, essential components of angiogenesis. To study MT dynamics, we used high-resolution fluorescence microscopy coupled with computational image analysis of fluorescently-labeled MT plus-ends to investigate MT growth dynamics and the regulation of EC branching morphology and directional migration. Time-lapse imaging of living Human Umbilical Vein Endothelial Cells (HUVECs) was performed following transfection with fluorescently-labeled MT End Binding protein 3 (EB3) and Mitotic Centromere Associated Kinesin (MCAK)-specific cDNA constructs to evaluate effects on MT dynamics. PlusTipTracker software was used to track EB3-labeled MT plus ends in order to measure MT growth speeds and MT growth lifetimes in time-lapse images. This methodology allows for the study of MT dynamics and the identification of how localized regulation of MT dynamics within sub-cellular regions contributes to the angiogenic processes of EC branching and migration.

Protocols for Human Umbilical Vein Endothelial Cell (HUVEC) culture, transient transfection of fluorescently-labeled markers of microtubule growth, live-cell imaging and automated analysis of interphase microtubule growth dynamics are detailed.

高分辨率成像定时，并在活体人脐静脉内皮细胞微管动力学自动分析

The physiological process by which new vasculature forms from existing vasculature requires specific signaling events that trigger morphological changes ...

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

We present an open source high content analysis instrument utilizing automated fluorescence lifetime imaging (FLIM) for assaying protein interactions using F&#246;rster resonance energy transfer (FRET) based readouts of fixed or live cells in multiwell plates. This provides a means to screen for cell signaling processes read out using intramolecular FRET biosensors or intermolecular FRET of protein interactions such as oligomerization or heterodimerization, which can be used to identify binding partners. We describe here the functionality of this automated multiwell plate FLIM instrumentation and present exemplar data from our studies of HIV Gag protein oligomerization and a time course of a FRET biosensor in live cells. A detailed description of the practical implementation is then provided with reference to a list of hardware components and a description of the open source data acquisition software written in &#181;Manager. The application of FLIMfit, an open source MATLAB-based client for the OMERO platform, to analyze arrays of multiwell plate FLIM data is also presented. The protocols for imaging fixed and live cells are outlined and a demonstration of an automated multiwell plate FLIM experiment using cells expressing fluorescent protein-based FRET constructs is presented. This is complemented by a walk-through of the data analysis for this specific FLIM FRET data set.

We present an open source high content analysis (HCA) instrument utilizing automated fluorescence lifetime imaging (FLIM) for assaying protein interactions using F&#246;rster resonance energy transfer (FRET) based readouts. Data acquisition for this openFLIM-HCA instrument is controlled by software written in &#181;Manager and data analysis is undertaken in FLIMfit.

开源高内涵分析利用自动荧光寿命成像显微镜

We present an open source high content analysis instrument utilizing automated fluorescence lifetime imaging (FLIM) for assaying protein interactions ...

Measuring the Confluence of iPSCs Using an Automated Imaging System

This study focuses on understanding how growing iPSCs on different ECM coating substrates can affect cell confluence. A protocol to assess iPSC confluence in real time has been established without the need to count cells in single cell suspension to avoid any growth perturbation. A high-content image analysis system was used to assess iPCS confluence on 4 different ECMs over time in an automated manner. Different analysis settings were used to assess cell confluence of adherent iPSCs and only a slight difference (at 24 and 48 hours with laminin) has been observed whether a 60, 80 or 100% mask was applied. We also show that laminin lead to the best confluence compared to Matrigel, vitronectin and fibronectin.

The goal of the protocol is to compare different extracellular matrix (ECM) coating conditions to assess how differential coating affects the growth rate of induced pluripotent stem cells (iPSCs). In particular, we aim to set up conditions to obtain optimal growth of iPSC cultures.

使用自动成像系统测量 iPSC 的汇合度。

This study focuses on understanding how growing iPSCs on different ECM coating substrates can affect cell confluence. A protocol to assess iPSC confluence ...