目前的工作部分由欧洲委员会支持, Horizon2020 项目 HCO-06-2015, "SmokeFreeBrain," GA 681120。

所有实验都是按照1975年《赫尔辛基宣言》中所概述的原则进行的, 这是2000年修订的, 并得到大学伦理委员会的批准. 
 1. 设备安装和记录状态控制 <ol> <li> 邀请参与者坐在计算机屏幕前的舒适的椅子上. </li>
	<li> 清洁额头上的皮肤 (如有必要, 轻轻地移动头发)、耳垂和用洗必泰消毒液的手指来消毒皮肤并去除皮肤上堆积的最浅的脂质层。 46 . </li> <li> 根据10-20 国际系统 (AFz、FPz、AF5、AF6、AF7、AF8) 47 , 将六电极 EEG 带应用到参与者和 #39 的前额。
	<li> 放置参考和接地电极, 每个耳垂一个. </li>
	<li> 将脉冲血到拇指上以记录 HR. </li>
	<li> 要收集 GSR, 请在非手的第二和第三个手指上放置两个电极。将电极放在第二方阵的手掌侧 (按照已经发布的程序) 48 . </li>
	<li> 检查 EEG 信号的阻抗, 使其低于 10 k 和 #8486; 通过在采集软件的接口上使用相应的阻抗测量按钮. 
	<ol> <li> 在 #34; 通道和 #34; 购置软件的设置, 设置和 #34; 采样率和 #34; 用于脑电图信号采集和 #34; 250 赫兹. #34; </li> </ol> </li>
	<li> 通过 eeg 放大器系统获取 eeg 信号, 并与 eeg 数据获取相关的 eeg 记录软件 (请参阅 资料表 )。通过按软件接口上的红色录制按钮启动 EEG 信号记录. </li>
	<li> 通过验证软件接口上是否存在适当的波形, 检查是否正确获取了 GSR 和 HR 信号. 
	<ol> <li> 在 #34; 通道和 #34; 购置软件的设置, 将 GSR 和 HR 信号采集的采样率设置为和 #34; 128 赫兹. #34; </li> </ol> </li>
	<li> 通过多用途测量能力系统获取自主信号 (即, GSR 和 HR) (请参阅材料 表 )。
	<li> 启动 GSR 和 HR 信号记录, 按下 #34; 记录和 #34; 相应的软件接口上的按钮. </li>
	<li> 请参与者查看白色 #34; 十字 #34; 黑色背景, 在屏幕上显示1分钟. 同时获取休眠状态活动, 用相对标记按钮指示其开始和结束的脑电图采集软件. </li>
	<li> 指示参与者, 配备脑电图、HR 和 GSR 记录仪, 以观看 PSA 视频, 在此期间, 信号录制发生。此外, 请参与者限制任何移动, 并在录制期间尽可能放松. </li>
	<li> 在 PSA 视频开始时, 按 EEG 采集软件接口上的起始标记按钮。在 PSA 视频的末尾按结束标记按钮. </li>
</ol> 2。刺激 <ol> <li> 选择目标刺激, 最好是根据预定义的关键性能指标 (kpi). </li>
	<li> 在设计实验协议时, 在由两个块组成的视频中, 散置六目标禁烟刺激 ( 即, 三点和三图像属于三选定的公益). 
	<ol> <li> 生成斑点块, 如下所示: 一个点基线 ( 即, 一个持续1分钟的记录片), 一列10禁烟点 (最多使用10项, 以类似于典型的电视广告插播), 以及现场基线. </li>
		<li> 生成图像块的方式如下: 图像基线 ( 即, 从国际情感图片系统 (IAPS) 数据库中取出的 8 中性图像 49 ), 一列10禁烟图像 (用于与斑点块的一致性),和图像基线. </li>
	</ol> </li> <li> 在参与者和 #39 中避免定位效果的偏差; 反应, 首先显示一半的参与者的斑点块和一半的参与者的图像块. 
	为了避免特定项目级别的位置偏差, 在每个块中以随机顺序显示刺激. </li>
	<li> 选择适当的1分钟视频基线 ( 即, 尽可能地保持中性, 如纪录片摘录), 并将其放置在视频协议中的斑点块之前和之后立即. </li>
	<li> 选择适当的图像基线 ( 即 尽可能中性, 如从 IAPS 数据库中取出的8中性图像) 49 , 并将其立即放置在视频协议中的图像块之前和之后. </li>
	<li> 在设计图像块时, 将每个图像的显示持续时间设置为9秒, 以确保检测到自主组件中的最终变化, 这是与 EEG 信号相比较慢的响应。在每个图像集之间, 在黑屏上显示一个白色十字, 以重建中心固定点. </li>
</ol> 3。信号处理 注意: 以下计算步骤可与各种公开可用的信号处理计算软件一起执行, 如 EEGLAB 50 或 LEDALAB 51 , 用于分析 GSR 信号。虽然在这里使用了专门开发的软件进行一些计算, 但下面介绍的步骤并不取决于所使用的特定软件。因此, 以下步骤将无法精确地描述与计算软件的交互, 而是将显示实现结果所需的逻辑步骤. 
<ol> <li> 脑电图信号处理 
	<ol> <li> 要检测和删除由于眼睛移动、闪烁和肌肉工件而导致的组件, 请应用陷波滤镜 (50 hz)、带通滤波器 (2-30 hz) 和独立分量分析 (ICA) 52 过程到 EEG 跟踪。
		<ol> <li> 使用陷波器 (50 hz) 过滤 EEG 信号, 以拒绝主电源组件, 然后使用带通滤波器 (2-30 hz) 来拒绝与正在调查的认知过程无关的频率分量. </li>
			<li> 在 ICA 中转换 EEG 时间序列 52 . </li>
			<li> 查找并删除与工件相关的独立组件. 
			注意: 与眼部工件相关的独立成分与大脑成分的形状和大小明显区别。一旦检测到, 在重组时间域中的信号之前, 从 ICA 程序中删除与眼部工件相关的组件. </li>
			<li> 使用保留的独立组件重组时间域中的 EEG 信号. </li>
		</ol> </li> <li> 对于每个主题, 从静止状态估计单个 alpha 频率 (宇航联合会), 根据科学家中建议的方法定义感兴趣的频带。ific 文献 28 . 
		注意: 这一步是重要的, 因为每个主题的频率波段的定义都必须单独确定. </li>
		<li> 为每个皮层区域和每个主题计算全局场强 (GFP) 16 。
		<ol> <li> 在感兴趣的频率波段 (特别是θ (IAF-6:IAF-2) 和 alpha (IAF-2:IAF+2)) 中筛选 EEG 信号, 根据在科学文献中建议的定义 28 . </li>
			<li> 计算 GFP 16 作为来自特定电极的 EEG 信号对皮层感兴趣区域的总和 (, 如左、右前额叶皮层。过滤在某一波段和平均值由被考虑的电极的数量。请参见以下等式: <img alt="Equation 1" src="/files/ftp_upload/55872/55872eq1.jpg"/> </li> </ol> </li> </ol> </li> <li> 方法退出索引 注: 在几项研究中, 额叶皮质已被表明作为一个感兴趣的领域的方式或退出的态度, 响应广泛的刺激范围 53 , 54 , 55 , 56 , 57 , 58 。
	<ol> <li> 应用将 aw 索引定义为 aw = GFPa_right-GFPa_left 的公式, 其中 GFPa_right 和 GFPa_left 表示从右侧 (AF6 和 AF8) 和左 (AF5 和 AF7) 电极在 alpha 波段 59 中计算的 GFP, 类 = "xref" > 60 </li> <li> 估计每秒的 AW 脑指数的波形, 然后在刺激期间的平均值. </li>
		<li> 根据在实验开始和结束时获得的基线 EEG 活动来标准化 AW 索引. 
		注意: 正值 aw 值意味着对主体所表达的刺激的方法动机, 而负 aw 值表示退出趋势. </li>
	</ol> </li> <li> 努力索引 注: 几项研究展示了人脑在人类各种任务中的研究和应用 28 , 33 。
	<ol> <li> 用于评估脑部的工作, 在θ带中使用额电极 (AFz, FPz, AF5, AF6, AF7, AF8) 28 , 11 , 33 , 45 . </li> <li> 从前端电极估计 GFP 16 。根据在实验开始和结束时获得的基线脑电图活动, 将努力指数数据标准化, 如 AW 指数. </li>
		<li> 对于结果解释, 请记住, 较高的工作级别索引意味着更高的任务难度级别 31 . </li>
	</ol> </li> <li> 情绪索引 <ol> 获取 HR 信号, 请使用泛汤普金斯算法 <li> 61 。
		<li> 使用恒定电压法 (0.5 V) 62 获取皮肤电导. </li>
		<li> 使用 LEDAlab 软件 67 获取皮肤电导的主音成分 ( 即 s 健电导水平 (SCL))。
		<ol> <li> 选择和 #34; 连续分解分析和 #34; 从 "分析" 菜单中. 
			注意: 建议通过单击 #34 优化分析; 优化和 #34; 按钮. </li>
			<li> 单击和 #34; 应用和 #34; 接受分析并绘制分解. </li>
		</ol> </li> <li> 为了匹配 SCL 和 hr 信号, 请参阅影响计划的 circumplex 模型 63 , 64 , 其中空间点的坐标分别由 HR 定义 (水平轴) 描述的价和由 SCL (垂直轴) 描述的唤醒现象 36 . </li>
		<li> 若要获取 monodimensional 变量, 请使用 ei 10 (由公式定义) 所描述的主题的情绪状态: ei = 1-#946;/和 #960; #39;. 
		注意: 在这里, <img alt="Equation 2" src="/files/ftp_upload/55872/55872eq2.jpg"/> hr 和 <img alt="Equation 3" src="/files/ftp_upload/55872/55872eq3.jpg"/> 构成了 #918;-分别对 hr 和 GSR 进行了评分变量. img alt ="公式 5" src = "/文件/ftp_upload/55872/55872eq5. jpg"/>> 以弧度表示, 计算为 arctang (<img alt="Equation 4" src="/files/ftp_upload/55872/55872eq4.jpg"/> <img alt="Equation 3" src="/files/ftp_upload/55872/55872eq3.jpg"/>)。因为在 [和 #8722; 1, 1] 之间的角度会有所不同, 所以 EI 在 [和 #8722; 1, 1] 之间会有所不同. </li>
		<li> 对于结果解释, 请记住, 较高的 EI 值意味着参与者所经历的更积极的情绪, 反之亦然. </li>
	</ol>
	</li>
</ol>

作者没有什么可透露的。

修改协议和疑难解答:
这里描述的协议可以被修改以用于不同的实验情况。特别是, 这里提供的刺激的列车可以改变成一个系列仅基于图像或仅在视频。此更改将允许分别测试仅打印或仅视频广告。然而, 这里没有特别的变化, 需要的步骤, 信号处理, 因为发生的眼部文物将发生在任何情况下, 他们必须从脑电图信号删除。当仅显示视频时, 基线只能包含视频, 而不使用从 IAPS 数据库中拍摄的图像 (如上所述)。一个常见的情况下, 需要故障诊断所有脑电图记录是存在的几个肌肉文物由于主题运动期间的录音会议。在这种情况下, 重要的是要监测收集的脑电图跟踪的质量, 并检查录像会话的视频, 以检查是否发生这样的运动文物后。如果出现此类工件, 则需要对应用 ICA 的过程进行修改, 以消除此类工件。由于随机发生的这些文物沿着 EEG 的痕迹, ICA 的程序可以很容易地产生一个组成部分, 这种肌肉制品存在, 并可以消除他们从连续的 counter-transformation 在时间域返回干净的脑电图数据.
技术的局限性:
在这里描述的大脑和情感记录技术有几个限制, 需要考虑, 当它适用于实验程序不同于一个提出。特别是, ICA 程序是敏感的数字脑电图传感器使用。因此, 减少用于加速录制的传感器数量将影响可能删除的工件数。事实上, 这里描述的过程包括六传感器, 并允许删除两个主要的工件组件 (例如,眼部和毛肌制品)。使用的 EEG 传感器数量越少, 可以删除的各种工件的数量就越少。
所提出的技术的另一个局限性是与大脑记录的总时间有关。事实上, 在类似的研究中建议的总持续时间为每个科目最多1小时, 以避免因疲倦或厌烦而引起的可能的混淆效应。在这个程度上, 刺激的长度必须受到严格的控制, 并且一组广泛的刺激不能在同一参与者身上进行测试, 或者至少不能在一次运行中进行。在1小时的订婚期间, 参与者必须仔细了解该研究, 并要求签署的知情同意, 配备实验仪器, 并记录在休息条件下, 在实验任务的会议。为了避免超过建议的时间限制, 一个可能的故障排除考虑是让两个实验者同时设置仪器。
关于现有方法的意义:
与传统的广告评价技术相比, 本文提出的记录和分析公益的大脑和情绪反应的协议具有一些优势。特别是, 其中之一是在 compilating 口头调查表时, 使用比所需的更低的样本量来获得显著结果的可能性。事实上, 虽然这里描述的协议可以用20-30 参与者的样本大小进行卓有成效的实施, 但口头调查表需要至少100受访者。
此外, 目前的技术可以研究生理反应不依赖于测试参与者的口头报告, 提供更多的本能反应的刺激接触。
未来应用:
拟议的议定书可适用于较大的主题样本, 以收集关于公益的有效性的进一步信息。一个未来的应用是确定特定的人口目标的先验预测的功能定义公益的有效性, 传达适当的健康信息到每个人口细分
协议中的关键步骤:
协议中的关键步骤包括适当选择基线, 以评估所提供的刺激的重要性, 以及工件拒绝。在这里, 我们使用了 IAPS 数据库, 它提供了大量的图像, 其情感反应被评估为基线, 和 ICA 的方法, 消除了由眼球和肌肉运动诱发的 EEG 痕迹的伪影。
获得的数据的意义:
在任何指数中, 性别因素没有统计学意义的影响, 表明在禁烟刺激的反应中, 年轻男性和女性没有任何区别。这似乎是与抽烟的习性的相当同类的传播一致在意大利人口在被调查的年龄人口之中74。
关于 AW 指数, "获奖" 运动的结果是, 比起 "有效的" 和 "无效的" 公益, 对图像和斑点刺激的态度明显更接近。这可以用象征性的通信方式来解释 "授予" PSA。此外, 该点类别的统计显着的影响, 以及相同的趋势的图像类别, 是符合的结果, 在前面的研究, 在右或左半球激活-反映的不同 P300振幅变化-被曝光的中性, 促进和对比吸烟消息刺激75。
关于努力指数, "有效" 的形象是最努力的。这可以用一个非常清晰的信息来解释, 那位接受气管切开术的女士的形象传达了一个侧面的句子 "不要告诉人们吸烟是坏的, 给他们看。相反, "被授予的" 形象, 描绘一支香烟象征着动脉充分的脂肪沉积, 获得了最高的努力水平, 可能由于较不为人所知的吸烟习惯的影响, 在一般人群中比较著名的肺部效应。最后, "无效" 的形象, 描绘了一个人与他的儿子后面的句子 "思考, 不吸烟", 可能引起了相当高的努力值由于文字和图片之间的关系显然很差, 可能诱使观察者尝试了解这两个元素之间的连接。与 "有效" 图像相反, 属于同一运动的视频报告了最高的努力价值, 可能是由于叙述故事的复杂性, 其中一个年轻人讲述了他刚45岁时所受到的心脏病发作。, 以及这一事件的所有后果, 从他胸口的伤疤到他日常生活中的局限性。这一结果是根据最近的一项研究提供的证据表明, 在视频广告的叙事结构的存在导致更高的θ功率在左正面的76。
最后, "有效" 运动显示了最高的 EI 值, 可能是由于病人的移情作用引起的estimonials 的运动, 因为显性的消息。这一结果可以与我们的样本 (16-19 岁) 在同一年龄组的问卷中获得的证据相一致。调查表比较了恐惧诱导的禁烟图像的感知与更积极的形象的感知, 它表明, 参与者倾向于恐惧诱导一个, 因为它比第一个77更 "吸引人"。此外, 为 "有效" 运动估计的较高 EI 值, 与先前关于禁烟电视广告的试点研究11相一致, 其证据证明, 情绪化的反吸烟广告与与其他类型的广告78相比, 在成年人口中, 以退出尝试次数来计算的有效性提高了。
选择了一个年轻的人口样本, 因为有关意大利烟草消费启动的流行病学数据主要涉及15-17 岁的人74。在本研究中取得的结果, 虽然对中学生的有限样本进行, 但支持这种 neurometric 指数评估禁烟 PSA 评价的效用。在 SmokeFreeBrain 研究计划的框架内, 对更大的学生样本的进一步研究, 可能会为年轻人的 "有效" 公益的产生提供更具决定性的信息。在目标人群和所公布的消息之间有一个合适的存在, 这就增加了消息对内隐态度和相关行为的影响, 并对提升的消息79产生了更积极的评价。最后, 在这个框架中, 设计一个有效的运动所感兴趣的主要因素之一是选择最合适的见证80。显然, 这项调查需要扩展到一个较老的样本, 因为对广告的生理、认知和自主反应, 特别是禁烟公益在不同年龄段的人群中可能有不同27,81 ,82。最后, 现在和将来的研究结果将有可能减少公共资金的浪费, 与产生和广播 "无效的" 公益不同的目标人群有关。

正如奥美强烈指出的, "人们不去想他们的感受, 不要说出他们的想法, 不要做他们所说的。因此, 可以推断人类不是理性的存有。这一假设进一步得到证明, 许多有关经济问题的决定不受直接意志控制, 而是依赖于自动、快速和有效的认知过程1。此外, 情感机制可以影响这些决策过程, 有助于执行特定的操作2,3,4。在这种情况下, 在市场营销研究中使用的传统工具5中, 研究能够提供有关消费者行为的其他信息的技术变得更加重要。这项调查仍然依赖于客户对产品和广告信息的感知/评估, 口头表达和自我报告, 这是一种易受多种偏见影响的方法。事实上, 到目前为止, 用于了解新产品或服务的市场表现的典型方法仅基于自我和焦点组。然而, 社会心理学和市场研究的研究已经认识到, 自我是不可靠的准确预测客户的偏好6。上述研究的作者对鞋类零售业的售前预测进行了一项研究, 确定 self-report-based 方法在预测成功方面的准确性很差, 而脑部数据则达到了预期准确度为80%。消费者神经科学是一个研究领域, 作为对这一需求的答案, 并由典型的神经科学方法的应用, 与市场和经济交流有关的人类行为的调查7。"神经" 一词指的是神经工具的应用-如眼球跟踪, 皮肤传导, 心率, 脑电图, 和功能磁共振成像 (fMRI)-根据不同公司的需求进行市场研究设计8,9. 上述技术正在吸引来自不同公司的越来越多的兴趣: 在过去的十年中, 神经公司的崛起是令人印象深刻的7。神经, 实际上, 允许调查与商业有关的几个方面: 广告目标定义在性别方面10,11, 区域性12,13,广告本身10,14,15, 年龄16, 品牌解决方案17, 价格体系结构18, 场景定位和扬声器的性别19, 客户购买态度20, pre-retail 测试6。
关于 EEG, 两个指数被用来评估受广告刺激的对象所返回的反应: AW 和努力指数。第一个是植根于戴维森 (1990) 的研究, 这表明, 在两个大脑半球活动的正面不对称意味着不同的动机倾向的建议刺激。特别是, 一些作者21,22报告了两个截然不同的神经系统的存在, 调解与方法相关的动机和与撤退相关的动机, 在左和右半球局部化,分别.根据这一证据, 前额叶皮质 (PFC) 在电路中起着重要的作用, 它既能实现正负激励。具体来说, 左和右 PFC 的差异参与, 随着左前激活的积极动机和增加右侧前激活在消极动机, 已报告的各种研究23,24,25. 此外, 各种报告13、14突出显示了应用 AW 索引的可能性--定义为两个半球之间的 alpha 波段中的前额活动的差异--到广告领域。除了对这一主题进行广泛的调查外, 还对特定的产品类别进行了研究, 例如香水10, 其中女性报告了广告的积极动机, 而男性则报告了负面的动机。对 AW 索引的研究也考虑了一些实验性的样本特征, 例如主题是否为广告项目26的使用者。例如, 在对啤酒商业的反应中, 啤酒饮用者比不喝啤酒的受试者报告的 PFC 活化水平更高。此外, 被考虑的另一个参数是参与者的性别。例如, 为了观察家庭汽车的位置, 男性 (不同于女性) 在整个持续时间27中对商业表现出积极的动机。研究的最后一个参数是实验对象的年龄。一个令人振奋的高潮与一些 "不太好" 的图像更大脑欣赏的年轻人比成人27。综上所述, 从α额不对称获得的 AW 指数得出的阳性值表明, 左活动更高, 暗示了对刺激的积极动机。反之, 负值表示更高的正确活动, 暗示消极的动机来回应刺激。
第二脑电图指数, 大脑的努力 (CE), 定义为额θ活动在 PFC, 已被描述在一些研究。特别是, 这些研究显示了 CE 的高值如何与更高的任务难度级别28相链接。这在 neuroaesthetics 被特别调查了, 响应听觉文学刺激29;在航空电子学30;在不同的监听条件下31,32;和人机交互33。
根据来自不同应用领域的经验证据的建议, 情感参与是刺激记忆的催化剂。这一现象也在神经中被调查, 表明在商业广告的观察过程中, 积极或消极的情绪处理是形成稳定记忆痕迹的一个重要因素34。此外, 值得考虑的是, 一个 "无意识的情绪" 发生时, 大脑系统调解无意识核心 "喜欢" (在其他皮层下地区, 伏隔核及其连接) 没有结合有意识的意识35. 在本篇文章的开头, 人们强调了对广告刺激的感知和反应常常是无意识的。神经的目的之一是调查情感反应的这方面。通过匹配 GSR 和 HR 来创建自主索引, 因为已知这两个自主参数反映了对刺激的情感响应36。由此产生的 EI 是概念化的基础上的扬抑模型的影响, 理论的罗素和巴雷特5。在这个模型中, HR 以图形方式呈现在 x 轴上, 而 GSR 在 y-axis 上, 返回关于刺激的价 (正或负) 和唤醒 (低或高激活) 的信息, 分别为37,38。EI 已经应用于听觉文学刺激的测试29, 以及电视广告的10,16,19,39。因此, 作者认为, 将 EI 应用到公益, 特别是反吸烟公益是值得的。
每年, 全世界有600万人因吸烟而死亡, 面对这一紧急情况的主要工具包括禁烟公益40。这种 PSA 自二十世纪五十年代以来就已经播出, 但对一般公众的效用是非常多变的, 导致了一系列有效或无效的运动。一般而言, 禁烟运动的成效, 是根据宣传运动后公共卫生的上升而量度的;态度、信念或行为的改变 (即提高认识, 改变消极态度, 增加退出线路的次数,等等);媒体的影响 (即,积极的讨论和对社会媒体的赞赏);等。此外, 通常采用标准经济评估设计, 如 cost-effectiveness 分析、成本效用分析或成本效益分析41。最后, 除了 in-depth 提供的证据外, 专门的 cost-effectiveness 评估, 通过比较运动费用和节省的医疗费用, 可以肯定有效的运动节省了一些资金, 并防止了数十数以千计的过早死亡42,43,44。
因此, 对特定公益的大脑/情绪反应似乎值得通过神经技术来进行调查, 以获得对更传统的显式反馈进行补充的生理反应的测量。访谈和问卷调查。
本研究报告所采用的指数已被应用于禁烟公益的参与者和地点的减少样本中, 支持神经技术应用于神经调查的潜在用处。公益11,45中的有效性功能。
本研究的目的是提出一系列的方法步骤, 可以导致准确地测量禁烟公益的大脑和情绪知觉。这一措施是初步分析的本能和生理基础和特征的特定 PSA, 是有效或无效的产生公众的反应。

<table>
	<tbody>
		<tr>
			<td>BrainVision Engineering, LiveAmp</td>
			<td>Brain Products GmbH 
			Zeppelinstrasse 7 
			82205 Gilching Germany</td>
			<td>BP-200-3000</td>
			<td>32 Channel wireless EEG amplifier system</td>
		</tr>
		<tr>
			<td>BrainVision Engineering, Neuromarketing Headset</td>
			<td>Brain Products GmbH 
			Zeppelinstrasse 7 
			82205 Gilching Germany</td>
			<td>N/A still a prototype</td>
			<td>6 channel EEG headset, Prototype system for EEG measurements to conduct neuromarketing studies</td>
		</tr>
		<tr>
			<td>BrainVision Recorder</td>
			<td>Brain Products GmbH 
			Zeppelinstrasse 7 
			82205 Gilching Germany</td>
			<td>BP-170-3000</td>
			<td>EEG recording Software for EEG data acquisition</td>
		</tr>
		<tr>
			<td>Nexus-10 MKII</td>
			<td>Mind Media BV Louis Eijssenweg 2B 6049CD Herten The Netherlands</td>
			<td>NX10B-8000mAh SN 0939130157</td>
			<td>Biofeedback and Neurofeedback 10 channels acquiring system multipurpose measuring capabilities (heart rate, skin conductance and more)</td>
		</tr>
		<tr>
			<td>BioTrace+</td>
			<td>Mind Media BV Louis Eijssenweg 2B 6049CD Herten The Netherlands</td>
			<td></td>
			<td>Recording software for Nexus-10 MKII data acquisition</td>
		</tr>
	</tbody>
</table>

electroencephalographic, heart rate, and galvanic skin response assessment for an advertising perception study: application to antismoking public service announcements

对广告、产品和包装的评估传统上是通过基于自我和焦点组的方法进行的, 但是这些方法在科学方面往往显得不准确。神经科学越来越多地应用于对商业刺激的感知和反应的神经基地的调查, 以支持传统的营销方法。在这方面, 某一部门或市场由公共服务公告 (公益) 代表。本协议的目的是应用脑电图 (eeg) 和自主信号分析来研究对选择的禁烟公益的反应。使用两个脑电图指数: 额α带脑电图不对称 (方法提取 (AW) 指数) 和额θ (努力指数)。此外, 还计算了自电皮肤反应 (GSR) 和心率 (HR) 信号得出的自主情绪指数 (EI)。本议定书描述了一系列操作和计算步骤, 通过上述指标正确估计一组受试者对选定数量的禁烟公益的情绪和大脑反应。特别是, 以象征性的沟通方式 (根据各专门委员会收到的奖品归类为 "授予的") 的运动获得了最高的方法价值, 按 AW 指数估计。一个地方和一个图像属于相同的 PSA 运动的基础上, "恐惧激起上诉", 并与叙事/经验沟通的方式 (分类为 "有效" 的基础上, 经济/健康相关的改进促进) 报告最低和最高的努力值分别。这可能是由于讲故事 (点) 的复杂性和图像的即刻 (一位接受过气管切开手术的女士)。最后, 同样的 "有效" 运动显示了最高的 EI 值, 可能是因为显性的见证和信息的传递所引起的共鸣。

22健康科目 (平均年龄17.64 ±0.95 岁, 范围 = 16-19 岁) 在自愿基础上登记。关于吸烟的习惯, 7 的参与者不是吸烟者, 9 是轻吸烟者 (每天5香烟), 6 是重的吸烟者 (和 #62; 每天5支香烟)。吸烟者的参与者报告说, 他们的第一支烟在平均年龄为13.38 ±3.01 岁, 而且没有一个参与者报告过戒烟。所有受试者都获得了有关该研究的详细信息, 并签署了知情同意书。这项实验是根据1975年《赫尔辛基宣言》所概述的原则进行的, 该项修订于 2000年, 并获大学伦理委员会批准。
在本研究中, 我们使用了考夫曼65和 Varcoe66所建议的 kpi, 并选择了三目标禁烟市场活动。其中有两项是在被 classificatied 为 "有效" 和 "无效" 的基础上列入的, 这是关于促进健康和提高一般人口的经济状况的官方数据所证明的。第三个 PSA 被归类, 并根据从各专门委员会获得的赞赏, 以所收到的奖金数额表示。具体而言, 三选定的禁烟公益在本研究中为: i) cdc (图像: cdc; 斑点: cdc 罗斯福)67, 美国 2012-2015, 有效的68,44, 引起恐惧的吸引力和叙事/经验沟通方式;二) 认为不吸烟 (图像和视频)69, 菲利普莫里斯, 美国 1998, 无效的70,71, 家长式沟通方式;和 iii) 脂肪香烟 (图像和视频)72, 英国心脏基金会, 英国 2003, 授予 (克里欧奖2005电视和无线电公共服务铜牌, 投资促进机构效力奖2004获奖者金表;投资促进机构效力奖2004优胜者最佳的奉献为有效性优胜者;和3帕尔马非盟艺术节国际 de la Publicite 戛纳 2004), 象征性的沟通风格。
统计分析包括一个方差因素, PSA 的类别作为一个内部因素与三水平 (有效, 无效, 并且被授予) 和性别 (M/F) 作为一个相互作用。吸烟态度不包括在分析中, 因为在三可能的群体 (即,不吸烟者, 轻吸烟者, 和重吸烟者) 的人数较少, 但在这三组的参与者的均匀分布允许分析在折叠样本中的性能。重要的是要回顾, 方差分析测试有足够的统计能力来处理相对较少的参与者, 如本研究中的73, 前提是因素的数量低于 4, 如本例所示。
方法提取指数:
对于图像, 没有发现三战役之间有统计学意义的差异 (F (240) = 2.649, p = 0.083), 但报告的 "获奖" 图像的 AW 值高于报告的 "无效" PSA (图 1,左)。
对于视频刺激, 发现了当场类别 (即,有效, 无效, 或授予) 的统计学意义上的效果 (F (240) = 3.171, p = 0.050)。事后分析突出了增加的 AW 值估计为 "被授予的" 点与 "有效" 一个 (p = 0.047) 和一个类似强的倾向 (p = 0.060) 比较 "无效" 点 (图 1, 右)。
总之, 属于 "获奖" 运动的图像和点都获得了最积极的方法值, 按 AW 指数估计。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55872/55872fig1.jpg"> 
图 1: 选定的禁烟 PSA 市场活动的 AW 索引估计.左边是与图像相关的结果, 右边是与 "有效"、"无效" 和 "授予" 禁烟公益的斑点有关的结果。误差线表示标准偏差。<a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55872/55872fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
努力指数:
方差分析测试突出了该类别 (即,有效, 无效, 或授予) 变量的统计显着的效果的两个图像 (f (240) = 8.589, p = 0.001) 和现货 (f (240) = 5.441, p = 0.008) 刺激。事后分析显示, 对于图像, "有效的" 一个明显低于 "无效" (p = 0.009) 和 "授予" (p 和 #60; 0.001) 部分 (图 2, 左)。此外, 对点进行的事后分析表明, 为 "有效" 视频报告的努力值比 "授予的" 点 (图 2, 右) 的估计要高得多 (p = 0.003)。
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55872/55872fig2.jpg"> 
图 2: 选定的禁烟 PSA 市场活动的工作量索引估计.左边是与图像相关的结果, 右边是与 "有效"、"无效" 和 "授予" 禁烟公益的斑点有关的结果。误差线表示标准偏差。<a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55872/55872fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
情绪指数:
一般而言, 报告的 "有效" 运动的 EI 值高于 "无效" 和 "已授予" 的禁烟 PSA 图像和斑点。关于图像刺激, 虽然在评估的条件之间缺乏显著的统计学差异, 但在 "有效" 图像上增加 EI 值的强烈趋势与 "无效" 相比是可以理解的 (图 3, 左)。对于视频刺激, 发现了斑点分类因子的统计学意义上的影响 (F (232) = 3.978, p = 0.029)。此外, 事后分析显示, 与 "有效" (p = 0.013) 相比, "无效" 点的 EI 值下降, 与 "授予的" 点 (图 3, 右) 相比, 具有明显相似的趋势 (p = 0.060)。一般而言, 报告的 "有效" 运动的 EI 值高于 "无效" 和 "被授予" 的价值, 无论是对图像还是现场tismoking 公益
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/55872/55872fig3.jpg"> 
图 3: 选定的禁烟 PSA 市场活动的 EI 估计.左侧的结果与图像有关, 右侧的结果与 "有效"、"无效" 和 "授予" 禁烟公益的斑点有关。误差线表示标准偏差。<a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55872/55872fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Electroencephalographic, Heart Rate, and Galvanic Skin Response Assessment for an Advertising Perception Study: Application to Antismoking Public Service Anno

Electroencephalographic, Heart Rate, and Galvanic Skin Response Assessment for an Advertising Perception Study: Application to Antismoking Public Service Announcements

下面的协议描述了一系列的操作和计算步骤, 以正确估计一个群体的情绪和大脑反应, 对选定数量的公共服务公告 (公益) 吸烟, 播出美国和欧洲在期间在1998和2015之间。

脑电图, 心率和电皮肤反应评估为广告知觉研究: 应用到禁烟公共服务公告

The evaluation of advertising, products, and packaging is traditionally performed through methods based on self-reports and focus groups, but these ...

electroencephalographic-heart-rate-galvanic-skin-response-assessment

Nanyang Technological University

Research

JoVE Journal

Neuroscience

14.2K 次观看.  Sapienza University of Rome. 下面的协议描述了一系列的操作和计算步骤, 以正确估计一个群体的情绪和大脑反应, 对选定数量的公共服务公告 (公益) 吸烟, 播出美国和欧洲在期间在1998和2015之间。

视频: Electroencephalographic, Heart Rate, and Galvanic Skin Response Assessment for an Advertising Perception Study: Application to Antismoking Public Service Announcements

Creating Objects and Object Categories for Studying Perception and Perceptual Learning

In order to quantitatively study object perception, be it perception by biological systems or by machines, one needs to create objects and object categories with precisely definable, preferably naturalistic, properties1. Furthermore, for studies on perceptual learning, it is useful to create novel objects and object categories (or object classes) with such properties2.
Many innovative and useful methods currently exist for creating novel objects and object categories3-6 (also see refs. 7,8). However, generally speaking, the existing methods have three broad types of shortcomings.
First, shape variations are generally imposed by the experimenter5,9,10, and may therefore be different from the variability in natural categories, and optimized for a particular recognition algorithm. It would be desirable to have the variations arise independently of the externally imposed constraints.
Second, the existing methods have difficulty capturing the shape complexity of natural objects11-13. If the goal is to study natural object perception, it is desirable for objects and object categories to be naturalistic, so as to avoid possible confounds and special cases.
Third, it is generally hard to quantitatively measure the available information in the stimuli created by conventional methods. It would be desirable to create objects and object categories where the available information can be precisely measured and, where necessary, systematically manipulated (or 'tuned'). This allows one to formulate the underlying object recognition tasks in quantitative terms.
Here we describe a set of algorithms, or methods, that meet all three of the above criteria. Virtual morphogenesis (VM) creates novel, naturalistic virtual 3-D objects called 'digital embryos' by simulating the biological process of embryogenesis14. Virtual phylogenesis (VP) creates novel, naturalistic object categories by simulating the evolutionary process of natural selection9,12,13. Objects and object categories created by these simulations can be further manipulated by various morphing methods to generate systematic variations of shape characteristics15,16. The VP and morphing methods can also be applied, in principle, to novel virtual objects other than digital embryos, or to virtual versions of real-world objects9,13. Virtual objects created in this fashion can be rendered as visual images using a conventional graphical toolkit, with desired manipulations of surface texture, illumination, size, viewpoint and background. The virtual objects can also be 'printed' as haptic objects using a conventional 3-D prototyper.
We also describe some implementations of these computational algorithms to help illustrate the potential utility of the algorithms. It is important to distinguish the algorithms from their implementations. The implementations are demonstrations offered solely as a 'proof of principle' of the underlying algorithms. It is important to note that, in general, an implementation of a computational algorithm often has limitations that the algorithm itself does not have.
Together, these methods represent a set of powerful and flexible tools for studying object recognition and perceptual learning by biological and computational systems alike. With appropriate extensions, these methods may also prove useful in the study of morphogenesis and phylogenesis.

We describe a novel methodology for creating naturalistic 3-D objects and object categories with precisely defined feature variations. We use simulations of the biological processes of morphogenesis and phylogenesis to create novel, naturalistic virtual 3-D objects and object categories that can then be rendered as visual images or haptic objects.

创建对象和对象类别学习的认知与知觉学习

In order to quantitatively study object perception, be it perception by biological systems or by machines, one needs to create objects and object ...

科研

神经科学

Using Affordable LED Arrays for Photo-Stimulation of Neurons

Standard slice electrophysiology has allowed researchers to probe individual components of neural circuitry by recording electrical responses of single cells in response to electrical or pharmacological manipulations1,2. With the invention of methods to optically control genetically targeted neurons (optogenetics), researchers now have an unprecedented level of control over specific groups of neurons in the standard slice preparation. In particular, photosensitive channelrhodopsin-2 (ChR2) allows researchers to activate neurons with light3,4. By combining careful calibration of LED-based photostimulation of ChR2 with standard slice electrophysiology, we are able to probe with greater detail the role of adult-born interneurons in the olfactory bulb, the first central relay of the olfactory system. Using viral expression of ChR2-YFP specifically in adult-born neurons, we can selectively control young adult-born neurons in a milieu of older and mature neurons. Our optical control uses a simple and inexpensive LED system, and we show how this system can be calibrated to understand how much light is needed to evoke spiking activity in single neurons. Hence, brief flashes of blue light can remotely control the firing pattern of ChR2-transduced newborn cells.

Adult-born neurons expressing ChR2 can be manipulated in slice electrophysiological preparations in order to examine their contribution towards the function of olfactory neural circuits.

使用光刺激神经元的负担得起的LED阵列

Standard slice electrophysiology has allowed researchers to probe individual components of neural circuitry by recording electrical responses of single ...

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

Local interneurons are continuously regenerated in the olfactory bulb of adult rodents1-3. In this process, called adult neurogenesis, neural stem cells in the walls of the lateral ventricle give rise to neuroblasts that migrate for several millimeters along the rostral migratory stream (RMS) to reach and incorporate into the olfactory bulb. To study the different steps and the impact of adult-born neuron integration into preexisting olfactory circuits, it is necessary to selectively label and manipulate the activity of this specific population of neurons. The recent development of optogenetic technologies offers the opportunity to use light to precisely activate this specific cohort of neurons without affecting surrounding neurons4,5. Here, we present a series of procedures to virally express Channelrhodopsin2(ChR2)-YFP in a temporally restricted cohort of neuroblasts in the RMS before they reach the olfactory bulb and become adult-born neurons. In addition, we show how to implant and calibrate a miniature LED for chronic in vivo stimulation of ChR2-expressing neurons.

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

Adult-born neurons of the olfactory bulb can be optogenetically controlled using Channelrhodopsin2-expressing lentiviral injection in the rostral migratory stream and chronic photostimulation with an implanted miniature LED.

慢性成人出生的嗅觉神经元的选择性病毒转导在体内 Optogenetic刺激

Local interneurons are continuously regenerated in the olfactory bulb of adult rodents1-3. In this process, called adult neurogenesis, neural stem cells ...

MPI CyberMotion Simulator: Implementation of a Novel Motion Simulator to Investigate Multisensory Path Integration in Three Dimensions

Path integration is a process in which self-motion is integrated over time to obtain an estimate of one's current position relative to a starting point 1. Humans can do path integration based exclusively on visual 2-3, auditory 4, or inertial cues 5. However, with multiple cues present, inertial cues - particularly kinaesthetic - seem to dominate 6-7. In the absence of vision, humans tend to overestimate short distances (&lt;5 m) and turning angles (&lt;30&deg;), but underestimate longer ones 5. Movement through physical space therefore does not seem to be accurately represented by the brain.
Extensive work has been done on evaluating path integration in the horizontal plane, but little is known about vertical movement (see 3 for virtual movement from vision alone). One reason for this is that traditional motion simulators have a small range of motion restricted mainly to the horizontal plane. Here we take advantage of a motion simulator 8-9 with a large range of motion to assess whether path integration is similar between horizontal and vertical planes. The relative contributions of inertial and visual cues for path navigation were also assessed.
16 observers sat upright in a seat mounted to the flange of a modified KUKA anthropomorphic robot arm. Sensory information was manipulated by providing visual (optic flow, limited lifetime star field), vestibular-kinaesthetic (passive self motion with eyes closed), or visual and vestibular-kinaesthetic motion cues. Movement trajectories in the horizontal, sagittal and frontal planes consisted of two segment lengths (1st: 0.4 m, 2nd: 1 m; &plusmn;0.24 m/s2 peak acceleration). The angle of the two segments was either 45&deg; or 90&deg;. Observers pointed back to their origin by moving an arrow that was superimposed on an avatar presented on the screen.
Observers were more likely to underestimate angle size for movement in the horizontal plane compared to the vertical planes. In the frontal plane observers were more likely to overestimate angle size while there was no such bias in the sagittal plane. Finally, observers responded slower when answering based on vestibular-kinaesthetic information alone. Human path integration based on vestibular-kinaesthetic information alone thus takes longer than when visual information is present. That pointing is consistent with underestimating and overestimating the angle one has moved through in the horizontal and vertical planes respectively, suggests that the neural representation of self-motion through space is non-symmetrical which may relate to the fact that humans experience movement mostly within the horizontal plane.

An efficient way to gain insight into how humans navigate themselves in three dimensions is described. The method takes advantage of a motion simulator capable of moving observers in ways unattainable by traditional simulators. Results confirm that movement in the horizontal plane is underestimated, while vertical movement is overestimated.

MPI Cyber​​Motion模拟器:一种新型的运动模拟器的执行情况调查三个维度的多感官的路径积分

Path integration is a process in which self-motion is integrated over time to obtain an estimate of one's current position relative to a starting point 1. ...

Local Application of Drugs to Study Nicotinic Acetylcholine Receptor Function in Mouse Brain Slices

Tobacco use leads to numerous health problems, including cancer, heart disease, emphysema, and stroke. Addiction to cigarette smoking is a prevalent neuropsychiatric disorder that stems from the biophysical and cellular actions of nicotine on nicotinic acetylcholine receptors (nAChRs) throughout the central nervous system. Understanding the various nAChR subtypes that exist in brain areas relevant to nicotine addiction is a major priority.
Experiments that employ electrophysiology techniques such as whole-cell patch clamp or two-electrode voltage clamp recordings are useful for pharmacological characterization of nAChRs of interest. Cells expressing nAChRs, such as mammalian tissue culture cells or Xenopus laevis oocytes, are physically isolated and are therefore easily studied using the tools of modern pharmacology. Much progress has been made using these techniques, particularly when the target receptor was already known and ectopic expression was easily achieved. Often, however, it is necessary to study nAChRs in their native environment: in neurons within brain slices acutely harvested from laboratory mice or rats. For example, mice expressing "hypersensitive" nAChR subunits such as &alpha;4 L9&prime;A mice 1 and &alpha;6 L9&prime;S mice 2, allow for unambiguous identification of neurons based on their functional expression of a specific nAChR subunit. Although whole-cell patch clamp recordings from neurons in brain slices is routinely done by the skilled electrophysiologist, it is challenging to locally apply drugs such as acetylcholine or nicotine to the recorded cell within a brain slice. Dilution of drugs into the superfusate (bath application) is not rapidly reversible, and U-tube systems are not easily adapted to work with brain slices.
In this paper, we describe a method for rapidly applying nAChR-activating drugs to neurons recorded in adult mouse brain slices. Standard whole-cell recordings are made from neurons in slices, and a second micropipette filled with a drug of interest is maneuvered into position near the recorded cell. An injection of pressurized air or inert nitrogen into the drug-filled pipette causes a small amount of drug solution to be ejected from the pipette onto the recorded cell. Using this method, nAChR-mediated currents are able to be resolved with millisecond accuracy. Drug application times can easily be varied, and the drug-filled pipette can be retracted and replaced with a new pipette, allowing for concentration-response curves to be created for a single neuron. Although described in the context of nAChR neurobiology, this technique should be useful for studying many types of ligand-gated ion channels or receptors in neurons from brain slices.

In this paper, we describe a useful method to study ligand-gated ion channel function in neurons of acutely isolated brain slices. This method involves the use of a drug-filled micropipette for local application of drugs to neurons recorded using standard patch clamp techniques.

局部应用药物研究烟碱型乙酰胆碱受体功能的小鼠脑片

Tobacco use leads to numerous health problems, including cancer, heart disease, emphysema, and stroke. Addiction to cigarette smoking is a prevalent ...

A Proboscis Extension Response Protocol for Investigating Behavioral Plasticity in Insects: Application to Basic, Biomedical, and Agricultural Research

Insects modify their responses to stimuli through experience of associating those stimuli with events important for survival (e.g., food, mates, threats). There are several behavioral mechanisms through which an insect learns salient associations and relates them to these events. It is important to understand this behavioral plasticity for programs aimed toward assisting insects that are beneficial for agriculture. This understanding can also be used for discovering solutions to biomedical and agricultural problems created by insects that act as disease vectors and pests. The Proboscis Extension Response (PER) conditioning protocol was developed for honey bees (Apis mellifera) over 50 years ago to study how they perceive and learn about floral odors, which signal the nectar and pollen resources a colony needs for survival. The PER procedure provides a robust and easy-to-employ framework for studying several different ecologically relevant mechanisms of behavioral plasticity. It is easily adaptable for use with several other insect species and other behavioral reflexes. These protocols can be readily employed in conjunction with various means for monitoring neural activity in the CNS via electrophysiology or bioimaging, or for manipulating targeted neuromodulatory pathways. It is a robust assay for rapidly detecting sub-lethal effects on behavior caused by environmental stressors, toxins or pesticides.
We show how the PER protocol is straightforward to implement using two procedures. One is suitable as a laboratory exercise for students or for quick assays of the effect of an experimental treatment. The other provides more thorough control of variables, which is important for studies of behavioral conditioning. We show how several measures for the behavioral response ranging from binary yes/no to more continuous variable like latency and duration of proboscis extension can be used to test hypotheses. And, we discuss some pitfalls that researchers commonly encounter when they use the procedure for the first time.

The Proboscis Extension Response (PER) conditioning protocol, developed for the honey bee (Apis mellifera), provides an ecologically-relevant and easily quantifiable means for studying several different mechanisms of learning in many insect species.

长鼻扩展响应协议为侦查行为可塑性的昆虫:应用基础，生物医学和农业研究

Insects modify their responses to stimuli through experience of associating those stimuli with events important for survival (e.g., food, mates, threats). ...

Flat Mount Imaging of Mouse Skin and Its Application to the Analysis of Hair Follicle Patterning and Sensory Axon Morphology

Skin is a highly heterogeneous tissue. Intra-dermal structures include hair follicles, arrector pili muscles, epidermal specializations (such as Merkel cell clusters), sebaceous glands, nerves and nerve endings, and capillaries. The spatial arrangement of these structures is tightly controlled on a microscopic scale - as seen, for example, in the orderly arrangement of cell types within a single hair follicle - and on a macroscopic scale - as seen by the nearly identical orientations of thousands of hair follicles within a local region of skin. Visualizing these structures without physically sectioning the skin is possible because of the 2-dimensional geometry of this organ. In this protocol, we show that mouse skin can be dissected, fixed, permeabilized, stained, and clarified as an intact two dimensional object, a flat mount. The protocol allows for easy visualization of skin structures in their entirety through the full thickness of large areas of skin by optical sectioning and reconstruction. Images of these structures can also be integrated with information about position and orientation relative to the body axes.

Mammalian skin contains a diverse array of structures - such as hair follicles and nerve endings - that exhibit distinctive patterns of spatial organization. Analyzing skin as a flat mount takes advantage of the 2-dimensional geometry of this tissue to produce full-thickness high-resolution images of skin structures.

小鼠皮肤及其毛囊图案和感觉轴突形态分析中的应用平山成像

Skin is a highly heterogeneous tissue. Intra-dermal structures include hair follicles, arrector pili muscles, epidermal specializations (such as Merkel ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method described here enables the investigator to measure long-lasting (from minutes to hours) high-quality intracellular responses from single Drosophila R1-R6 photoreceptors and Large Monopolar Cells (LMCs) to light stimuli. Because the recording system has low noise, it can be used to study variability among individual cells in the fly eye, and how their outputs reflect the physical properties of the visual environment. We outline all key steps in performing this technique. The basic steps in constructing an appropriate electrophysiology set-up for recording, such as design and selection of the experimental equipment are described. We also explain how to prepare for recording by making appropriate (sharp) recording and (blunt) reference electrodes. Details are given on how to fix an intact fly in a bespoke fly-holder, prepare a small window in its eye and insert a recording electrode through this hole with minimal damage. We explain how to localize the center of a cell's receptive field, dark- or light-adapt the studied cell, and to record its voltage responses to dynamic light stimuli. Finally, we describe the criteria for stable normal recordings, show characteristic high-quality voltage responses of individual cells to different light stimuli, and briefly define how to quantify their signaling performance. Many aspects of the method are technically challenging and require practice and patience to master. But once learned and optimized for the investigator's experimental objectives, it grants outstanding in vivo neurophysiological data.

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Sharp microelectrodes enable accurate electrophysiological characterization of photoreceptor and visual interneuron output in living Drosophila. Here we show how to use this method to record high-quality voltage responses of individual cells to controlled light stimulation. This method is ideal for studying neural information processing in insect compound eyes.

为记录细胞内电压响应电法<em&gt;果蝇</em&gt;光感受器和中间对光刺激<em&gt;在体内</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network dynamics in neuronal cultures. In contrast with more traditional methods, such as patch-clamping, which can only record activity from a single cell, MEAs can record simultaneously from multiple sites in a network, without requiring the arduous task of placing each electrode individually. Moreover, numerous control and stimulation configurations can be easily applied within the same experimental setup, allowing for a broad range of dynamics to be explored. One of the key dynamics of interest in these in vitro studies has been the extent to which cultured networks display properties indicative of learning. Mouse neuronal cells cultured on MEAs display an increase in response following training induced by electrical stimulation. This protocol demonstrates how to culture neuronal cells on MEAs; successfully record from over 95% of the plated dishes; establish a protocol to train the networks to respond to patterns of stimulation; and sort, plot, and interpret the results from such experiments. The use of a proprietary system for stimulating and recording neuronal cultures is demonstrated. Software packages are also used to sort neuronal units. A custom-designed graphical user interface is used to visualize post-stimulus time histograms, inter-burst intervals, and burst duration, as well as to compare the cellular response to stimulation before and after a training protocol. Finally, representative results and future directions of this research effort are discussed.

Mouse neuronal cells cultured on multi-electrode arrays display an increase in response following electrical stimulation. This protocol demonstrates how to culture neurons, how to record activity, and how to establish a protocol to train the networks to respond to patterns of stimulation.

时间依赖性增加对微电极阵列刺激神经元细胞培养物的网络响应

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network ...

Quantifying Infra-slow Dynamics of Spectral Power and Heart Rate in Sleeping Mice

Three vigilance states dominate mammalian life: wakefulness, non-rapid eye movement (non-REM) sleep, and REM sleep. As more neural correlates of behavior are identified in freely moving animals, this three-fold subdivision becomes too simplistic. During wakefulness, ensembles of global and local cortical activities, together with peripheral parameters such as pupillary diameter and sympathovagal balance, define various degrees of arousal. It remains unclear the extent to which sleep also forms a continuum of brain states-within which the degree of resilience to sensory stimuli and arousability, and perhaps other sleep functions, vary gradually-and how peripheral physiological states co-vary. Research advancing the methods to monitor multiple parameters during sleep, as well as attributing to constellations of these functional attributes, is central to refining our understanding of sleep as a multifunctional process during which many beneficial effects must be executed. Identifying novel parameters characterizing sleep states will open opportunities for novel diagnostic avenues in sleep disorders.
We present a procedure to describe dynamic variations of mouse non-REM sleep states via the combined monitoring and analysis of electroencephalogram (EEG)/electrocorticogram (ECoG), electromyogram (EMG), and electrocardiogram (ECG) signals using standard polysomnographic recording techniques. Using this approach, we found that mouse non-REM sleep is organized into cycles of coordinated neural and cardiac oscillations that generate successive 25-s intervals of high and low fragility to external stimuli. Therefore, central and autonomic nervous systems are coordinated to form behaviorally distinct sleep states during consolidated non-REM sleep. We present surgical manipulations for polysomnographic (i.e., EEG/EMG combined with ECG) monitoring to track these cycles in the freely sleeping mouse, the analysis to quantify their dynamics, and the acoustic stimulation protocols to assess their role in the likelihood of waking up. Our approach has already been extended to human sleep and promises to unravel common organizing principles of non-REM sleep states in mammals.

Here, we present experimental and analytical procedures to describe the temporal dynamics of the neural and cardiac variables of non-REM sleep in mice, which modulate sleep responsiveness to acoustic stimuli.

量化睡眠小鼠的光谱功率和心率的慢速动力学

Three vigilance states dominate mammalian life: wakefulness, non-rapid eye movement (non-REM) sleep, and REM sleep. As more neural correlates of behavior ...