这项研究得到新西兰视力远景研究基金会、健康促进机构、奥克兰大学师资研究发展基金和奥克兰大学医学院 PBRF 基金的支持。
我们感谢奥克兰大学 (香农男孩和 Harpreet 辛格) 的两名验光学学生, 他们完成了手工视频分析, 以检查眨眼响应测量中的协议。

这里描述的所有方法都已得到了北 B 健康和残疾伦理委员会的批准, 参考编号:13/关税壁垒/181。
1. 技术要求
<ol><li>使用能直接将空气吹向角膜的充气装置。这可以通过空气压缩机连接到空气罐上的软管来实现。为确保婴儿的安全, 请将其与可调压力调节器配合使用, 以限制气压和限制吹气时间。 注: 商用系统将压缩机纳入系统内, 不需要空气罐。</li>
	<li>使用可调节的头戴或帽子, 可以由婴儿佩戴, 并允许适当定位的空气粉扑管在眼睛旁边。</li>
	<li>设置两个可调音量设置的便携式音频扬声器。</li>
	<li>设计和设置设备的计算控制, 使音调和空气粉扑可以自动管理, 精确定时, 并在不同的时间间隔。</li>
	<li>设置至少60赫兹帧速率的便携式摄像机, 以手动记录上只是一眨眼响应。要测量响应时间, 请使用可以捕获试验开始的摄像机, 方法是记录发声音, 或者拍摄程序进入软件包的试验起始指示器。</li>
	<li>安装并运行任何视频编辑软件, 允许音频和视频组件 (如Adobe 首版 Pro) 的分帧步进, 以允许研究员测量刺激和眨眼响应之间的时间间隔。</li>
</ol>2. 实验范式
<ol><li>在10块中管理试验。每个块应包含以下试用类型。<ol>
<li>包括八配对试验, 其中每个学习试验包括一个750毫秒的音调 (80 分贝, 1 赫), 重叠和终止与 100-ms 空气粉扑 (~ 1/20 psi)9,10,11。</li>
		<li>包括一个未配对的试验, 这包括一个单一的750毫秒的语气单独提出在没有空气粉扑测试一个条件上只是一眨眼反应指示联想学习。</li>
		<li>包括一种体感试验, 由单一的100毫秒空气粉扑试验, 单独提出在没有的语气, 以测试角膜敏感性的空气粉扑和正确的定位头带。</li>
	</ol>
</li>
	<li>随机化连续试验之间的间隔 (例如, 8-十六年代, 平均试验时间间隔大约十二年代)9,10,11。</li>
	<li>在单个会话中测量连续五块 (即总共50个试验), 总会话时间约为12分钟。</li>
	<li>为了测试条件是否持续, 管理两个 EBC 的50次实验, 至少1小时。</li>
</ol>3. 测试程序
<ol><li>在一个适度点燃的房间 (约300莱克丝) 执行 EBC 在实验之间保持照明水平一致。</li>
	<li>将两个便携式扬声器放置在婴儿两侧, 大约40厘米从头部两侧。</li>
	<li>将定制的头带放在婴儿头部, 调整头部大小。</li>
	<li>将带有空气粉扑喷嘴和相邻红外线传感器的柔性管放置在头的轴向平面上, 使其与儿童右眼约 1-3 厘米 (图 1)。</li>
	<li>在开始实验之前, 检查实验设置是否优化以提供角膜粉扑。提供一个测试空气粉扑到婴儿的眼睛, 并观察是否有上只是一眨眼发生, 并在摄像机上可见。如果没有观察到上只是一眨眼, 调整头带位置, 使充气单位更接近眼睛。</li>
	<li>在实验期间记录婴儿的上只是一眨眼反应。定位摄像机, 使视野的方向朝向婴儿的眼睛。相机应该足够接近, 以辨别小眼睛的运动, 但不能如此接近, 使婴儿可以移动的框架外, 如果他/她变得挑剔。</li>
	<li>确保实验者对刺激时机有所掩饰。如果 EBC 用户界面指示下一次试验何时发生, 请让研究人员将自己定位在远离计算机界面上, 以便他们不知道何时会交付下一个刺激。这将阻止研究者对刺激本身做出反应, 并可能有助于婴儿的学习反应。</li>
</ol>4. 优化一岁婴儿的实验性设置
<ol><li>确保有两位研究人员可以进行实验。</li>
	<li>当把头带放在婴儿身上时, 请看护者把婴儿放在大腿上, 并朝侧面定位。这使得研究者能够从婴儿头部的侧面和后面接近, 而第二个研究者则与婴儿接触。</li>
	<li>放置头带和定位喷嘴应尽可能快速和冷静地执行, 以确保婴儿的依从性。将手放在喷嘴的尖端, 可防止婴儿在定位时将头部转向喷嘴。</li>
	<li>在整个实验过程中, 让婴儿在看护者的膝上。 注意: 虽然高级椅子可以在实验之间提供更一致的环境, 但这可能会增加许多婴儿的焦虑并减少依从性。</li>
	<li>对于在实验中倾向于干扰头带的婴儿, 请使用温和的分心。</li>
	<li>在实验中, 头带通常会移动。如果婴儿不再响应空气粉扑或头带已移动显着远离眼睛, 暂停实验和调整头带位置, 然后继续。</li>
	<li>鼓励孩子在实验中查找, 以提高上只是一眨眼运动的规模;这样, 上只是一眨眼检测就可以得到改善。 注意: 这可以通过将婴儿放在显示器上播放婴儿友好视频 (例如, 与音频关闭的年龄合适的卡通) 稍高于他们的视线来实现。</li>
</ol>5. 使用摄像机录制的上只是一眨眼检测
<ol><li>让两位研究人员独立定义每个实验的上只是一眨眼反应。</li>
	<li>使用视频编辑软件分析每一个试验的2000毫秒潜伏期后的试验开始。</li>
	<li>使用音频波形检测第一帧的试用开始。<ol>
<li>对于配对 (cs + 美国) 和未配对 (cs) 的试验, 定义从发声音开始的试验起始。</li>
		<li>对于只有美国 (空气粉扑) 的体感试验, 请执行事先测试以确保摄像机的麦克风能够检测到空气粉扑所产生的微妙的100毫秒噪音。否则, 在软件包中编程的试验起始指示器的视频记录将有助于检测吹气的爆发。</li>
	</ol>
</li>
	<li>使用视频跟踪检测闪烁响应。记录眨眼潜伏期为以下两个: 眨眼发作-第一个框架, 眼睑开始关闭;眨眼峰-第一帧的眼睑是最大闭合。</li>
	<li>根据需要, 制定策略来处理模棱两可的反应。请参见下面的示例。<ol>
<li>合理排除部分眨眼, 这可能发生在眼睑不能完全关闭超过50%。</li>
		<li>排除的试验中, 音调/空气粉扑的开始是隐藏的外来噪音, 或在闪烁反应是模糊的视线, 从进一步的分析。</li>
		<li>在观察到多次眨眼的试验中, 分析了试验开始后的第一次眨眼反应。</li>
	</ol>
</li>
	<li>记录刺激开始和眨眼峰值之间的视频帧数。根据摄像机的采样率将滞后时间转换为实时。</li>
</ol>6. EBC 反应分类
<ol><li>在检测到上只是一眨眼响应发生的那些试验之后, 根据它与刺激开始的潜伏期 (图 2) 来定义眨眼类型。<ol>
<li>如果它发生在音后的前200毫秒间隔内, 则将眨眼定义为惊吓响应;这些代表了听觉音调的反射反应, 或闪烁的时间巧合地与音调开始, 将发生独立的空气粉扑。 注意: 可能会观察到对 (cs + 美国) 或未配对 (cs) 试验的惊吓反应, 在那里听觉音调被传递。</li>
		<li>如果它发生在空气粉扑的反应中, 将眨眼定义为体感反应;它表明角膜是否对空气粉扑敏感与当前头盔配置。这是一种无条件响应的类型 (见下文)。</li>
		<li>定义无条件反应作为感觉反应的空气粉扑在配对 (CS + 美国) 试验时发生650毫秒以上的音调开始。</li>
		<li>定义条件响应作为闪烁, 最佳定时与空气粉扑。它们在350到650毫秒之间开始, 在配对或未配对试验的音调出现后, 表明 CS 和美国之间的联想学习可能已经发生。</li>
		<li>如果在响应空气粉扑时没有发现闪烁, 则将试验定义为失败的响应;这些可以观察到配对 (CS + 美国) 试验或体感试验。这些反应表明, 空气粉扑没有达到眼睛。</li>
	</ol>
</li>
</ol>7. 联想学习的分析
注意: 以前已经描述了许多方法来评估联想学习9、10、11 (这里简要介绍)。研究人员应该修改这些方法或发展自己来定义是否发生了联想学习, 这取决于他们的实验设计。
<ol><li>首先, 排除没有收集可用数据的试验, 如具有技术错误的测试。</li>
	<li>通过计算所有可能的体感试验中上只是一眨眼反应的百分比来计算空气粉扑的灵敏度。</li>
	<li>通过比较10项试验连续块中条件反应的百分比, 评估实验过程中是否发生了联想学习。在实验过程中, 条件反应的百分比的增加可能表明已经发生了调节。 注: 或者, 建模上只是一眨眼潜伏期的变化可以用来评估婴儿在实验过程中是否适应了他们的反应时间。</li>
	<li>评估联合学习是否发生在实验性课程或两个实验组之间。使用任意标准来定义是否已发生调节 (例如, 在实验性会话中 > 40% 条件响应)6,9。</li>
</ol>

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作者没有什么可透露的。

修改和疑难解答: 商业或定制的上只是一眨眼调节系统应以实验控制的方式向主体提供刺激, 并能够检测行为反应。虽然这是一个非侵入性的程序, 但在不太顺从的人群 (如婴儿) 进行这些实验的技术要求是具有挑战性的。物理上连接头安装到眼睛是可能的动物实验, 例如调理实验绵羊12,13。年长的孩子可能会容忍佩戴带头巾的附气吹送单元6,7,8 , 可以鼓励保持一个恒定的眼睛位置使用电视作为分心。虽然在14的睡眠条件下, 非常年幼的婴儿的依从性是可以实现的, 但接近一岁的年长婴儿通常容易分心, 使他们成为一个独特的挑战人群。尽管有这些挑战, 上只是一眨眼的实验对于一些醒着的, 一岁的婴儿是可能的。我们的成功率类似于以前在文献中报道的 (40%-50% 合规)9, 成功的可能性可能反映在实验当天婴儿的气质。
议定书内的关键步骤: 我们建议适应一个商业上可用的 EBC 设置, 或使用一个定制的空气压力发生器如前所述, 为年轻的婴儿9,10,11。最初的挑战是在整个实验中保持空气吹送装置的正确位置, 这需要一个较小的, 修改过的头盔。眼睛安全是一个特别关注的不符合要求的婴儿试图消除头盔自己。因此, 必须在任何时候都密切监测婴儿。比例的儿童不会容忍戴头巾或接收空气粉扑的眼睛 (这里, ~ 35%), 虽然这可能会改善, 如果实验性的协议允许这种情况下, 轻轻地分散婴儿, 如果这不公然干扰婴儿行为。对于完成实验的孩子们来说, 只有气喷试验才能确认头盔正确定位, 因为在整个实验过程中, 空气粉扑的传递不佳可能导致调节反应的缺乏或逆转。
虽然手工视频分析耗时, 但目前难以实现婴儿上只是一眨眼反应的自动化措施。使用商用 EBC 设置的红外线传感器 (位于充气单元旁边的头带上) 记录从角膜到1赫取样速率的力矩反射, 通过测量反射率的变化来检测闪烁。在刺激开始之后。基线反射率的显著变化表明发生了上只是一眨眼, 自动分析可用于检测和分类闪烁响应。然而, 我们和其他9,10,11建议, 研究人员不应该依赖于红外角膜反射测量提供的商业 EBC 设置, 以检测眨眼在一岁的孩子。虽然这种方法可能适用于成人参与者或年龄较大的儿童6,7, 我们无法可靠地检测到在一岁的孩子的眨眼使用这种方法。在这里, 基线反射率的变化在试验纪元的长度上是可以忽略的, 这表明红外传感器没有检测到上只是一眨眼。此外, 假阳性峰值通常被观察到, 这是由婴儿的头部运动, 改变传感器的定位。在成人中进行的飞行员测试表明, 参与者必须专注于或稍微高于他们的水平视线, 与空气粉扑单位放置在 1-2 厘米的角膜, 以成功地检测 eyeblinks 的条件, 不能保持与婴儿一致。此外, 小 interpalpebral 裂隙大小的婴儿可能会降低角膜反射率, 使眨眼检测比成人或更大的儿童更困难。
对于现有方法的意义: 尽管需要手动视频摄像机分析和 60 Hz 视频的更低的时间分辨率相比, 1 赫反射率测量, 提高视频记录的成功率, 使这是一个优越的方法, 红外传感器。虽然角膜粉扑也提供了同样的头盔附件作为商业红外线传感器, 刺激交付似乎是更有弹性的定位比红外传感器, 因为空气粉扑强度可以根据需要调整。这避免了过度调整的头带, 这往往减少婴儿依从性。研究人员谁可以获得更高的成功率使用红外线传感器可能仍然需要支持摄像机分析。对于红外传感器记录的没有检测到的上只是一眨眼响应的试验, 视频记录可以区分真实的应答或无效的试验 (即, 儿童是否在正确的位置佩戴充气装置,它们是否没有闪烁, 或者红外线传感器是否检测不到闪烁)。同样, 录像是必要的, 以确认假阳性结果, 因为在审判期间的运动也可以产生伪装成眨眼的艺术品。眼球跟踪仪15,16和肌电图 9,10,14的眼轮匝肌是其他自动的眨眼检测方法, 超出了本文的范围,尽管没有对每个试验的空气粉扑单元定位进行目视确认, 但这些方法可能会与此处描述的红外线上只是一眨眼检测软件的缺点相同。
以前发表的眼睛眨眼调理工作集中在年轻的婴儿, 通常 4-5 月的年龄;这项工作增加了这一机构的文学通过描述与老年人使用的技术 (12, 1 月的年龄) 和独特的挑战, 这个年龄组提出。其他作者也使用了视频分析 (单独或结合与肌电图) 与婴儿9,10,11,14。本文的研究结果与先前的结果一致, 即帧间视频分析是检测婴儿 eyeblinks 的最佳方法, 因为肌电图和红外线监测都可能因面部和头部问题在非常年幼的孩子的运动。这项工作还支持的发现, 婴儿可以发展条件的反应, 即使在很小的年龄, 至少在650毫秒的延迟间隔9。
未来应用: 由于有限数量的研究试图上只是一眨眼调节实验的人婴儿, 在未来的实验中应仔细考虑刺激参数。在10天的17岁和至少在生命的第一个月内, 已经观察到了调理, 尽管在婴儿期的这些非常早期的阶段 (如1500 毫秒) 似乎更成功了..相比之下, 接近六月龄的儿童可以更可靠地适应较短的间隔10, 虽然比成人19的成功程度更低。650毫秒的语气和空气粉扑之间的间隔, 建议一岁的孩子在这个协议可以成功地诱导在4或5月大的婴儿9,10,11, 以及在老年人的调节孩子6,7。这表明, 上只是一眨眼调节是发育调节, 与最佳的延迟之间的语气和空气粉扑下降与婴儿年龄10, 虽然需要额外的实验, 以适应此协议, 以调查这些参数。
此技术的局限性: EBC 可能会有挑战性的设置, 并可能需要在每个单独的情况下进行故障排除。确定何时发生调理是很难定义的, 最好在多个试验或会话中观察到。例如, 其他研究在每块10试验中使用了40% 的任意调节率, 以定义调理, 并比较了连续实验课6、9、10之间的任何变化, 11。因此, 成功完成所需的试验次数, 以观察适应能力, 是很难实现的幼儿。
在数据分析方面, 特别的挑战之一是将刺激诱发的眨眼与自发 (内源) 闪烁9分开。个体间自发 eyeblinks 的速率不同, 但受环境因素的影响, 如室内湿度, 以及行为状态。双眨眼发生时, 有两个完整的眼睑关闭在一个非常短的时间空间 (例如, 400 毫秒)20, 这是可能的, 这些反映自发闪烁发生在同一时间内的反身性 (条件或无条件)上只是一眨眼.我们偶尔会在我们的数据集中观察到双闪烁, 并且, 使用此协议, 我们只记录了第一次闪烁的滞后时间。然而, 放弃这些试验可能增加确定性, 分析是捕捉刺激-诱发眨眼只。
研究人员还需要在延迟窗口中确定条件或无条件反应, 以及是否定义瞬发或眨眼峰值的潜伏期 (眼睑闭合)。在这里, 我们保守地定义了一个条件反应作为眨眼, 在650毫秒之前, 无论是配对或未配对的试验, 以提供确定性, 眨眼是启动之前, 空气粉扑开始, 而不是响应空气粉扑。但是, 此延迟窗口可以扩展为未配对的试验, 其中没有空气粉扑提出, 特别是如果使用眨眼峰值来定义响应的滞后时间。这里的重要原则是所有的事例都以同样的方式, 由所有考官来处理, 并且在开始进行数据分析之前确定这个标准。
总之, 帧帧视频分析可以提供一个可靠和可重现的方法, 评估经典的 EBC 反应的幼儿。这些措施提供了一种学习行为的衡量标准, 尽管需要全面的发展评估来充分描述神经发育在婴儿期的特点。

EBC 是一种经典的调理方式, 通常用于评估人类和其他动物的学习和记忆。传统的 EBC 范式指的是一种无害的 ' 条件 ' 刺激 (听觉音) 与 ' 无条件 ' 刺激 (一种诱发反身上只是一眨眼的角膜气泡) 之间的学会关联, 即无条件反应。联想学习是通过提出条件性的刺激和分类 eyeblinks 作为条件和无条件的反应-增加的条件反应数量随着时间的推移, 或超过一个会话, 可能表明联想学习正在发生。
上只是一眨眼调理范式可用于识别具有联想学习障碍的个体。延迟调节-条件性刺激 (CS) 开始之前, 但重叠的无条件刺激 (美国)-是由小脑电路1,2控制, 并在成人和儿童神经紊乱的干扰,包括精神分裂症3, 自闭症4,5, 和胎儿酒精频谱紊乱6,7。从临床的角度来看, 检查婴儿的上只是一眨眼调理可能支持早期发现和治疗这些神经疾病。然而, 虽然年龄较大的儿童可能会容忍6,7,8的角膜空气粉扑, 并可以保持一个稳定的头部/眼睛位置在实验期间, 婴儿不太顺从, 和传统的上只是一眨眼检测设备和软件不足。
在这里, 我们描述了一个简单的协议来检查一岁儿童的短时延迟上只是一眨眼调理, 这已经适应了以前在其他地方9,10,11的协议。该协议可以使用任何商业或定制的系统, 能够提供控制的空气喷到角膜。然而, 代替商用上只是一眨眼检测软件 (例如, 圣地亚哥仪器的上只是一眨眼检测系统, 它处理红外摄像机的信号), 我们和其他9,10,11,建议采用帧帧分析的摄像机记录来检测和分类一岁婴儿上只是一眨眼反应。

<table border="0" cellpadding="0" cellspacing="0" style="width:813px;" width="814">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Eyeblink conditioning system</td>
			<td style="width:204px;">San Diego Instruments</td>
			<td style="width:227px;">Model #2325-0145-W</td>
			<td style="width:167px;">includes portable airpuff unit and infrared emitter/receiver mounted to adjustable headband; eyeblink stimulus and response detection software; PC interface</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">4K Ultra HD action camera</td>
			<td style="width:204px;">RV77</td>
			<td style="width:227px;">Model SJ9000</td>
			<td>micro SD/TF card recording 90fps at 720p</td>
		</tr>
	</tbody>
</table>

classical short-delay eyeblink conditioning in one-year-old children

经典的上只是一眨眼调理 (EBC) 指的是有条件的刺激 (听觉音) 和无条件刺激 (对角膜的粉扑) 之间的关联。上只是一眨眼调理经常被用来检测小脑依赖性学习和记忆中的异常, 这是这类联想学习的基础。虽然成人和大龄儿童的实验使用商用设备比较简单, 但由于其依从性较差, 婴儿的上只是一眨眼调节更具挑战性, 这使得设备的正确定位变得困难。为了达到一岁婴儿的调理, 一个定制的或适应的商业系统可以用来向婴儿的角膜提供空气粉扑。主要的挑战在于成功地检测和分类行为反应。我们报告说, 自动眨眼检测方法在这个人群中是不可靠的, 并且应该通过对辅助摄像机记录的帧间分析来分析调节实验。该方法可用于研究上只是一眨眼条件下的发育变化, 并探讨这种模式是否能检测神经紊乱患儿。

一岁婴儿上只是一眨眼调节的可测性:
对一岁的婴儿来说, 检查上只是一眨眼条件所需的实验设置是很有挑战性的。在我们进行的实验中, 35% (11 名婴儿) 未能参加实验, 因为他们不会容忍戴头巾或接收空气粉扑到角膜 (n = 31 婴儿尝试)。大约一半 (52%, n = 16 婴儿) 完成或部分完成第一套50试验 (中位 [IQR] = 22.5 [0-50] 试验完成)。剩下的人参加了两次实验性会议。
取得可靠的实验结果需要在整个实验中正确定位头带和充气单元。对于那些参与实验的婴儿来说, 在大部分的试验中都可以获得空气粉扑 (77, 6% 的空气喷吹引起眨眼反应;n = 13 infants/557 尝试的充气粉扑试验)。其余的试验代表 "失败的试验", 其中婴儿不敏感的空气粉扑。
图 3说明了在50项试验的两个调节实验中吹气的成功。观察到的 eyeblinks 的数量被描述为空气粉扑试验的百分比 (即体感或配对试验)。眨眼反应从第一个婴儿下降的每一个连续的10块试验, 表明, 头带可能已分阶段的最佳位置, 在实验结束。相比之下, 第二个婴儿表明对空气粉扑的敏感性相对较好, 至少90% 的空气粉扑试验成功地引起了每一个后续的10个试验块的眨眼反应。
上只是一眨眼检测成功率使用摄像机分析: 手持式摄像机通过允许研究员手动跟踪婴儿的眼睛, 提供了一种可靠的捕获上只是一眨眼响应的方法。用人工帧间分析方法, 从90到3% 的试验中确定反应 (n = 13 infants/608 试验尝试)。其余的试验是指在拍摄过程中婴儿的眼睛被遮蔽的那些。
虽然视频分析需要主观的解释, 这个方法可以用来可靠地测量上只是一眨眼潜伏期从刺激发作。我们对两个独立观察者测量的眨眼响应时间的分析表明了良好的评分员的可靠性, 即测量的平均差为28.4 毫秒和95% 的协议限制 263.9 ms (图 4: n = 4实验分析了两个 observers/94 闪烁响应)。考虑到条件 eyeblinks 被定义为那些属于300毫秒的窗口, 这允许足够的精度来区分条件与无条件反应。注意, 那些有重大分歧的审判可以在观察者协商后很容易重新评估。
一岁婴儿上只是一眨眼调理: 图 5A和5B所示的两种调理实验说明了实验过程中对配对和未配对试验的上只是一眨眼反应潜伏期。第一个婴儿在第一个 EBC 会话中的上只是一眨眼反应在空气粉扑的开始和在定义条件响应的滞后时间窗口之外。此外, 这个婴儿没有对任何未配对的 (仅音) 试验作出反应。这表明, 这个婴儿在初次接触 EBC 范式时, 并没有学会将音调和气吹在一起。相比之下, 第二个婴儿, 谁是参加他们的第二次 EBC 会议, 例行眨眼之前, 空气粉扑开始, 并对所有未配对的试验作出反应, 表明婴儿已达到上只是一眨眼的条件后, 第一届会议50试验。图 5C说明了一种典型的方法, 用于评估6、9、10、11会话的多项试验中的联想学习。两个婴儿的条件反应的数量被描述在连续10个试验块和作为所有配对和未配对的试验的百分比在每个块。在这种情况下, 两个婴儿显示在实验期间联想学习的最小变化, 说明缺乏调理 (婴儿 1, 疗程 1) 或已经发生的调节 (婴儿 2, 会话 2)。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58037/58037fig1.jpg"> 图 1: 上只是一眨眼与一岁婴儿参加者建立了空调.这是一个定制的头带支持空气粉扑单位显示。<a href="https://www.jove.com/files/ftp_upload/58037/58037fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58037/58037fig2.jpg"> 图 2: 刺激和反应定义为一上只是一眨眼适应范例适合实验与一岁孩子.(a) 在配对试验中, 750 毫秒听音 (条件刺激 [CS]) 重叠, 并终止与 100 ms 空气粉扑 (无条件刺激 [美国])。眨眼反应的变化根据他们的时间相对于刺激发作 (惊吓反应: 0-200 ms; 条件反应 [CR]: 350-650 ms; 无条件响应 [UR]: 650-2000 ms)。响应定义与未配对 (CS) 试验相同。(B) 体感试验仅提供单100毫秒的空气粉扑 (美国), 而在300毫秒的窗口中发生的闪烁被归类为体感反应 (SSR)。<a href="https://www.jove.com/files/ftp_upload/58037/58037fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/58037/58037fig3.jpg"> 图 3: 一岁婴儿上只是一眨眼调理实验的例子, 说明空气粉扑灵敏度的变化.对空气粉扑的敏感性被描述为被观察的空气粉扑的反应的数量 (作为体感试验或作为配对的试验) 作为所有空气粉扑试验的百分比, 在以后每块10试验。<a href="https://www.jove.com/files/ftp_upload/58037/58037fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58037/58037fig4.jpg"> 图 4: 平淡的奥特曼剧情, 说明了两个独立观察者从摄像机录像中测量的眨眼响应时间的协议.灰色线表示测量之间的偏差 (定义为测量之间的平均值差), 以及协议的上限和下限 (即, 定义为平均差1.96 标准偏差)。<a href="https://www.jove.com/files/ftp_upload/58037/58037fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/58037/58037fig5.jpg"> 图 5: 一岁婴儿上只是一眨眼调理实验的例子, 说明条件反应的变化.闪烁响应时间显示为配对 (cs + 美国) 试验 (黑眼圈), 和不成对 (cs) 试验 (灰色方块)。灰色条表示空气粉扑的时间, 黑线表示 350-650 毫秒之间的滞后时间用于定义条件响应。在x轴上显示未能引出响应的试验。(A) 眨眼反应一般落在调节窗外, 未配对的试验未能引起上只是一眨眼反应。(B) 反应适当定时, 以配合空气粉扑的发生, 未配对的反应和配对的试验启动条件反应。(C) 实验过程中的调节被说明为每连续10次试验中有条件反应的数量 (被描述为所有配对和未配对试验的百分比)。<a href="https://www.jove.com/files/ftp_upload/58037/58037fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Classical Short-Delay Eyeblink Conditioning in One-Year-Old Children at JoVE.com

Classical Short-Delay Eyeblink Conditioning in One-Year-Old Children

该协议描述了一个适合于一岁婴儿实验的上只是一眨眼调理范式。商业或定制的设备可以用来提供刺激, 数据收集和分析应执行的视频记录。

经典短时滞上只是一眨眼在一岁儿童中的调节作用

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a ...

classical-short-delay-eyeblink-conditioning-in-one-year-old-children

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22.5K 次观看.  The University of Auckland. 该协议描述了一个适合于一岁婴儿实验的上只是一眨眼调理范式。商业或定制的设备可以用来提供刺激, 数据收集和分析应执行的视频记录。

视频: Classical Short-Delay Eyeblink Conditioning in One-Year-Old Children

Quantitative Assessment of Cortical Auditory-tactile Processing in Children with Disabilities

Objective and easy measurement of sensory processing is extremely difficult in nonverbal or vulnerable pediatric patients. We developed a new methodology to quantitatively assess children&#39;s cortical processing of light touch, speech sounds and the multisensory processing of the 2 stimuli, without requiring active subject participation or causing children discomfort. To accomplish this we developed a dual channel, time and strength calibrated air puff stimulator that allows both tactile stimulation and sham control. We combined this with the use of event-related potential methodology to allow for high temporal resolution of signals from the primary and secondary somatosensory cortices as well as higher order processing. This methodology also allowed us to measure a multisensory response to auditory-tactile stimulation.

Objective and easy measurement of sensory processing is extremely difficult in nonverbal or vulnerable pediatric patients. We developed a new methodology to quantitatively assess infants and children&#39;s cortical processing of light touch, speech sounds, and the multisensory processing of the 2 stimuli, without requiring active subject participation or causing discomfort in vulnerable patients.

皮质听觉，触觉加工的残疾儿童定量评价

Objective and easy measurement of sensory processing is extremely difficult in nonverbal or vulnerable pediatric patients. We developed a new methodology ...

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行为学

Trace Fear Conditioning in Mice

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings between a neutral stimulus and an unconditioned stimulus. There is a 20 sec trace period that separates each conditioning trial. On the following day freezing is measured during presentation of the conditioned stimulus (CS) and trace period. On the third day there is an 8 min test to measure contextual memory. The representative results are from mice that were presented with the aversive unconditioned stimulus (shock) compared to mice that received the tone presentations without the unconditioned stimulus. Trace fear conditioning has been successfully used to detect subtle learning and memory deficits and enhancements in mice that are not found with other fear conditioning methods. This type of fear conditioning is believed to be dependent upon connections between the medial prefrontal cortex and the hippocampus. One current controversy is whether this method is believed to be amygdala-independent. Therefore, other fear conditioning testing is needed to examine amygdala-dependent learning and memory effects, such as through the delay fear conditioning.

In the following experiment we describe a protocol for trace fear conditioning in mice. This type of associative memory includes a trace period that separates the neutral stimulus and the unconditioned stimulus.

跟踪恐惧条件小鼠

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings ...

Measurement of Fronto-limbic Activity Using an Emotional Oddball Task in Children with Familial High Risk for Schizophrenia

Adolescence is a critical developmental period where the early symptoms of schizophrenia frequently emerge. First-degree relatives of people with schizophrenia who are at familial high risk (FHR) may show similar cognitive and emotional changes. However, the neurological changes underlying the emergence of these symptoms remain unclear. This study sought to identify differences in frontal, striatal, and limbic regions in children and adolescents with FHR using functional magnetic resonance imaging. Groups of 21 children and adolescents at FHR and 21 healthy controls completed an emotional oddball task that relied on selective attention and the suppression of task-irrelevant emotional information. The standard oddball task was modified to include aversive and neutral distractors in order to examine potential group differences in both emotional and executive processing. This task was designed specifically to allow for children and adolescents to complete by keeping the difficulty and emotional image content age-appropriate. Furthermore, we demonstrate a technique for suitable fMRI registration for children and adolescent participants. This paradigm may also be applied in future studies to measure changes in neural activity in other populations with hypothesized developmental changes in executive and emotional processing.

This paper describes how to use the emotional oddball task and fMRI to measure brain activation in children and adolescents at familial high risk for schizophrenia (FHR).&#160;FMRI was used to measure differences in fronto-striato-limbic regions during an emotional oddball task. Children with FHR exhibited abnormal functional activation during adolescence.

的额边缘活度测量在儿童使用情绪穿着或行为古怪的任务与家族性高风险的精神分裂症

Adolescence is a critical developmental period where the early symptoms of schizophrenia frequently emerge. First-degree relatives of people with ...

Cortical Source Analysis of High-Density EEG Recordings in Children

EEG is traditionally described as a neuroimaging technique with high temporal and low spatial resolution. Recent advances in biophysical modelling and signal processing make it possible to exploit information from other imaging modalities like structural MRI that provide high spatial resolution to overcome this constraint1. This is especially useful for investigations that require high resolution in the temporal as well as spatial domain. In addition, due to the easy application and low cost of EEG recordings, EEG is often the method of choice when working with populations, such as young children, that do not tolerate functional MRI scans well. However, in order to investigate which neural substrates are involved, anatomical information from structural MRI is still needed. Most EEG analysis packages work with standard head models that are based on adult anatomy. The accuracy of these models when used for children is limited2, because the composition and spatial configuration of head tissues changes dramatically over development3.&#160;
In the present paper, we provide an overview of our recent work in utilizing head models based on individual structural MRI scans or age specific head models to reconstruct the cortical generators of high density EEG. This article describes how&#160;EEG recordings are acquired, processed, and analyzed with pediatric populations at the London Baby Lab, including laboratory setup, task design, EEG preprocessing, MRI processing, and EEG channel level and source analysis.&#160;

In recent years, there has been increasing interest in estimating the cortical sources of scalp measured electrical activity for cognitive neuroscience experiments. This article describes how high density EEG is acquired and how recordings are processed for cortical source estimation in children from the age of 2 years at the London Baby Lab.

高密度的脑电图记录在儿童的皮质来源分析

EEG is traditionally described as a neuroimaging technique with high temporal and low spatial resolution. Recent advances in biophysical modelling and ...

Measuring Attentional Biases for Threat in Children and Adults

Investigators have long been interested in the human propensity for the rapid detection of threatening stimuli. However, until recently, research in this domain has focused almost exclusively on adult participants, completely ignoring the topic of threat detection over the course of development. One of the biggest reasons for the lack of developmental work in this area is likely the absence of a reliable paradigm that can measure perceptual biases for threat in children. To address this issue, we recently designed a modified visual search paradigm similar to the standard adult paradigm that is appropriate for studying threat detection in preschool-aged participants. Here we describe this new procedure. In the general paradigm, we present participants with matrices of color photographs, and ask them to find and touch a target on the screen. Latency to touch the target is recorded. Using a touch-screen monitor makes the procedure simple and easy, allowing us to collect data in participants ranging from 3 years of age to adults. Thus far, the paradigm has consistently shown that both adults and children detect threatening stimuli (e.g., snakes, spiders, angry/fearful faces) more quickly than neutral stimuli (e.g., flowers, mushrooms, happy/neutral faces). Altogether, this procedure provides an important new tool for researchers interested in studying the development of attentional biases for threat.

Here we describe a touch-screen visual search paradigm that can be used to study threat detection across the lifespan. The paradigm has already been used in various studies demonstrating that both children and adults detect threatening stimuli like snakes, spiders, and angry faces faster than non-threatening stimuli.

测量注意偏向对威胁儿童和成人

Investigators have long been interested in the human propensity for the rapid detection of threatening stimuli. However, until recently, research in this ...

Quantifying Social Motivation in Mice Using Operant Conditioning

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered shuttle boxes were equipped with two operant levers (left and right) and a food receptacle in one chamber, which was then divided from the second chamber by an automated guillotine door covered by a wire grid. Different stimulus mice, rotated across testing days, served as a social stimulus behind the wire grid, and were only visible following the opening of the guillotine door. Test mice were trained to lever press in order to open the door and gain access to the stimulus partner for 15 sec. The number of lever presses required to obtain the social reward progressively increased on a fixed schedule of 3. Testing sessions ended after test mice stopped lever pressing for 5 consecutive minutes. The last reinforced ratio or breakpoint can be used as a quantitative measure of social motivation. For the second paradigm, test mice were trained to discriminate between left and right lever presses in order to obtain either a food reward or the social reward. Mice were rewarded for every 3 presses of each respective lever. The number of food and social rewards can be compared as a measurement of the value placed upon each reward. The ratio of each reward type can also be compared between mouse strains and the change in this ratio can be monitored within testing sessions to measure satiation with a given reward type. Both of these operant conditioning paradigms are highly useful for the quantification of social motivation in mouse models of autism and other disorders of social behavior.

This article describes a new protocol for the quantitative measurement of social motivation in mice using operant conditioning for a social reward. The protocol is useful for the measurement of social motivation in mouse models of autism and other disorders of social behavior.

量化的社会动机小鼠使用操作条件反射

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered ...

Whisker-signaled Eyeblink Classical Conditioning in Head-fixed Mice

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive repertoire of scientific techniques available to study learning and memory at the cellular level, it is ideal to have a stable cranial platform. Because mice do not readily tolerate restraint, they are usually trained while moving about freely in a chamber. Conditioned stimulus (CS) and unconditioned stimulus (US) information are delivered and eyeblink responses recorded via a tether connected to the mouse's head. In the head-fixed apparatus presented here, mice are allowed to run as they desire while their heads are secured to facilitate experimentation. Reliable conditioning of the eyeblink response is obtained with this training apparatus, which allows for the delivery of whisker stimulation as the CS, a periorbital electrical shock as the US, and analysis of electromyographic (EMG) activity from the eyelid to detect blink responses.

The preparation presented here for whisker-signaled eyeblink conditioning in head-fixed mice precisely stimulates specific whiskers while allowing mice to ambulate on a cylindrical treadmill. A whisker stimulation conditioned stimulus (CS) paired with a periorbital shock unconditioned stimulus (US) results in reliable associative learning on this apparatus.

在头固定小鼠胡须信号眨眼经典条件反射

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive ...

Psychophysical Tracking Method to Measure Taste Preferences in Children and Adults

The Monell two-series, forced-choice, paired-comparison tracking method provides a reliable measure of sweet taste preferences from childhood to adulthood. The method, which is identical for children, adolescents, and adults, is of short duration (&#60; 15 min), does not rely on sustained attention or place demands on memory (which would yield spurious age differences), and minimizes the impact of language development, making this method amenable to the cognitive limitations of pediatric populations. In this whole-mouth tasting method, subjects are asked to taste (without swallowing) pairs of solutions of different sucrose concentrations and to point to the solution they prefer. Each subsequent pair contains the participant&#39;s preceding preferred concentration and an adjacent stimulus concentration. The procedure continues until the subject chooses either a given concentration of sucrose when paired with both a higher and a lower concentration, or the highest or lowest concentration two consecutive times. Subjects are prevented from reaching response criteria on the basis of first or second position bias by the two-series design of the method, which counterbalances the order of solution presentation within each pair between the series (the weaker concentration is presented first in Series 1, second in Series 2). The geometric mean of the two sucrose concentrations chosen in Series 1 and 2 is an estimate of the participant&#39;s most preferred level of sucrose. Sucrose preference as determined with this laboratory-based measure has been shown to be associated with preference for sugars in foods and beverages and with taste receptor genotype, family history of alcoholism, and race/ethnicity, as well as depressive symptomatology among pediatric populations. The method has real-world relevance and has been applied to determine most preferred level of other tastes (e.g., salt), making it a valuable psychophysical tool.

Sweet taste has powerful hedonic appeal among people of all ages, particularly children. Described herein is a reliable and valid method that can be used to determine the level of sweetness most preferred, making it a valuable psychophysical tool for scientists.

心理跟踪方法来衡量儿童和成人的口味偏好

The Monell two-series, forced-choice, paired-comparison tracking method provides a reliable measure of sweet taste preferences from childhood to ...

Assessing Working Memory in Children: The Comprehensive Assessment Battery for Children &#8211; Working Memory (CABC-WM)

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of working memory in young school-age children. Working memory deficits have been identified in children with language-based learning disabilities, including dyslexia1,2 and language impairment3,4, but it is not clear whether these children exhibit deficits in subcomponents of working memory, such as visuospatial or phonological working memory. The CABC-WM is administered on a desktop computer with a touchscreen interface and was specifically developed to be engaging and motivating for children. Although the long-term goal of the CABC-WM is to provide individualized working memory profiles in children, the present study focuses on the initial success and utility of the CABC-WM for measuring central executive, visuospatial, phonological loop, and binding constructs in children with typical development. Immediate next steps are to administer the CABC-WM to children with specific language impairment, dyslexia, and comorbid specific language impairment and dyslexia.

Working memory predicts a significant amount of variance for a variety of cognitive tasks, including speaking, reading, and writing. However, few tools are available to assess working memory in children. We present an innovative, computer-based battery that comprehensively assesses different components of working memory in school-age children.

评估儿童工作记忆:儿童综合评估电池 - 工作记忆（CABC-WM）

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of ...

An Electrophysiology Protocol to Measure Reward Anticipation and Processing in Children

We present a protocol designed to measure the neural correlates of reward in children. The protocol allows researchers to measure both reward anticipation and processing. Its purpose is to create a reward task that is appropriate for young children with and without autism while controlling reward properties between two conditions: social and nonsocial. The current protocol allows for comparisons of brain activity between social and nonsocial reward conditions while keeping the reward itself identical between conditions. Using this protocol, we found evidence that neurotypical children demonstrate enhanced anticipatory brain activity during the social condition. Furthermore, we found that neurotypical children anticipate social reward more robustly than children with autism diagnoses. As the task uses snacks as a reward, it is most appropriate for young children. However, the protocol may be adapted for use with adolescent or adult populations if snacks are replaced by monetary incentives. The protocol is designed to measure electrophysiological events (event-related potentials), but it may be customized for use with eye-tracking or fMRI.

This protocol is designed to measure reward anticipation and processing in young children with and without autism. Specifically, the protocol is designed to study the neural correlates of reward during social and nonsocial conditions while controlling for reward between conditions.

儿童奖励预期与处理的电生理学协议

We present a protocol designed to measure the neural correlates of reward in children. The protocol allows researchers to measure both reward anticipation ...