本研究主要由 Lundbeck 基金会资助。
此外, Knud Eriksens Mindefond, Søster 弗纳 Lipperts 喜欢, Fonden Lægevidenskabens, Fremme Aage 路易喬安妮喜欢支持这项研究。

所有受试者必须在考试前给予书面同意, 录音协议必须由适当的地方道德审查委员会批准。这里描述的所有方法都得到了区域科学伦理委员会和丹麦数据保护局的批准。
注: 录音是用"TRONDNF"记录协议, 这是软件的一部分 (见材料表)。使用的其他设备是双极刺激器, 一个 50 Hz 的消噪装置, 一个功放, 和一个模拟到数字 (a/d) 板 (也建议是一个音频放大器的反馈的肌电 (肌电信号) 活动, 和一个静音盒, 以切断声音在电刺激)。用 MScan 方法估计的电机单位数包括三阶段: 1) 研究对象 (如神经兴奋性研究), 2) 记录 CMAP 扫描, 3) 用 MScan 软件分析结果。下面描述的记录程序特定于我们使用的软件和工具 (见材料表);这些将需要适应其他软件和硬件。
1. 专题的准备工作
<ol><li>屏幕上的主题, 以确保他们没有任何神经系统疾病 (特别是神经病和腕管综合征) 的历史, 除了将被调查的疾病组。</li>
	<li>对考试内容进行详细的指导, 并要求书面同意。<ol>
<li>通知主题, 当录音开始时, 功率将逐渐增加到最大值, 然后逐步减少, 考试将需要 5-6 分钟。</li>
		<li>解释这个主题会在手和手指上感受到发痒的感觉。</li>
		<li>如果该主题感到太不舒服, 请通知该主题, 在录制过程中随时可以立即关闭电源。</li>
	</ol>
</li>
	<li>用皮肤准备凝胶和酒精清洁主题的手和前臂。</li>
	<li>将活动记录电极放在拇指的 metacarpo-指骨关节上的外展拇短肌和参考电极上 (图 1)。</li>
	<li>将接地电极放在手背上。</li>
	<li>将这些电极连接到预放大器 (图 1)。</li>
	<li>将手指贴在一起, 以消除因自愿运动而产生的噪音和艺术品 (图 1)。</li>
	<li>保持32摄氏度和36摄氏度之间的皮肤温度与一盏暖灯。</li>
</ol>2. 记录 CMAP 扫描
注:下面描述的所有软件操作都是特定于我们使用的软件和工具 (见材料表);这些将需要适应其他软件和硬件。
<ol><li>启动半自动电脑系统。</li>
	<li>从 "选择记录协议" 窗体中选择"MScan"记录协议。</li>
	<li>接受前置放大器和刺激器的默认设置。</li>
	<li>在 "选择记录参数" 窗体上, 在 "输出文件" 框中输入一个2或3个字母的运算符前缀, 然后单击"ok"按钮。</li>
	<li>当程序显示原始的肌电信号输入时, 请手动选择"MScan 参数" (扫描步骤、interstimulus 间隔和刺激宽度), 或接受默认参数: 0.2ms 的刺激宽度、扫描步骤0.2% 和 interstimulus间隔为0.5 秒. 单击"确定"或按"转义"键继续。 注意: 本研究中接受了默认参数。过滤器设置为3赫兹-3 赫。</li>
	<li>将可复位双极刺激电极放在腕正中神经, 以找到最低阈值的位置。单击"确定"开始刺激。</li>
	<li>下一次显示器显示刺激 (最初是15% 最大输出) 和肌电信号响应;调整电极位置以找到最低阈值 (即最大响应)。如果有必要, 调整录音电极的位置, 以确保 CMAP 的形状双相一个峰值 (如果可能的话)。如果需要, 用"插入"和"删除"键调整刺激强度。然后, 单击"确定"继续.
</li>
	<li>"修改" 响应出现, "窗口", 其中的响应测量, 由水平洋红色线指示。确保窗口前面的短绿线 (表示基线) 是刺激工件和响应之间的平坦线。如有必要, 请通过按住鼠标右键设置窗口开始, 将光标向右拖动, 然后松开窗口结尾的按钮。 注: CMAP 峰值高度从基线到窗口中的上行峰值, 并由垂直蓝线表示。 单击"确定"继续。</li>
	<li>用非极化粘附刺激阴极电极取代可复位电极, 并将阳极2厘米下部沿正中神经;底部的痕迹现在显示高增益肌电信号。鼓励主题找到最轻松的位置, 他们的手, 以减少自发活动。单击"确定"继续。</li>
	<li>按"插入"键手动增加刺激强度, 直到刺激电流高于 CMAP 最大振幅的水平;首先, 增加3% 的刺激强度的步骤, 然后为精细调整使用步骤1%。</li>
	<li>在单击"确定"按钮开始 CMAP 扫描之前, 请检查窗口和峰值测量。</li>
	<li>请注意, 在对 20 supramaximal 刺激 (预扫描) 进行记录后, 刺激强度会在小步骤中自动减少, 从 supramaximal 刺激直到不再有可辨识的马达响应, 然后再设置 20 CMAPs(扫描后) 记录。 注意: 数据被绘制为"CMAP 扫描", 或者是一个详细的刺激响应曲线, 其振幅与 y 轴上的电机响应和 x 轴上的刺激强度有关。在健康的学科, 这创造了 S 形曲线, 而在减少电机单位数量的患者, 如在 ALS, 曲线发展一个阶梯状的外观 (图 2)。</li>
	<li>注意, 除了从静音盒中获得的反馈, 如果主题放宽, 可以在左侧的中间面板中进行检查。看看左上面板上的 CMAP (修改), 左下板, 刺激强度 (阈值) 的降低。按照右侧的 CMAP 扫描。</li>
	<li>单击右下角的"确定"按钮完成录制, 除非需要重复扫描并保存数据。 注意: 在完成 "图例和缩放" 窗体时, 必须替换所有问号。</li>
</ol>3. MScan 分析
注:下面描述的所有软件操作都是特定于我们使用的软件和工具 (见材料表);这些将需要适应其他软件和硬件。
<ol><li>
将模型拟合到 CMAP 扫描中
	<ol>
<li>要对记录进行离线分析, 请启动分析程序并单击"ok"以选择最后的记录进行分析.
</li>
		<li>单击 "从 MScanFit"菜单中的"选择适合 MScan QZD 文件" . 注意: 你可以选择"适合 MScan 在记忆文件"。如果 CMAP 扫描被记录在与此处使用的设备不同的情况下,则可以选择"在 dat (mA mV) 文件中调整 MScan"。请参见下面如何创建 DAT 文件。</li>
		<li>观察程序首先生成一个初步模型, 然后再进行优化。在 "优化" 框中的多色进度条完成且"停止"按钮变灰时, 不需要用户干预。</li>
		<li>初步模型提供了一个模型的第一个猜想, 从斜率和扫描连续部分的方差得出。请注意, 原始扫描 (黑色) 和从模型 (洋红色) 生成的扫描可以并排绘制。按照模型和改进在模型上的文本显示和替代显示。 注意: 可以根据 "数据绘制"、"绘制振幅为" 和 "绘图类型" 框中的选项随时选择可选显示。</li>
		<li>通过进行串行调整以尽量减少模拟和记录的 CMAP 扫描16之间的差异, 执行优化以改进对记录 CMAP 的匹配。 注意: 这会连续进行几个优化, 从初步模型开始, 如前文详细说明15所述。如果优化增加了单位数, 则下一次对扫描进行拟合的尝试将通过生成具有更多单元的模型开始, 而如果第一个优化减少了单位数, 则第二次尝试从具有较少单位的模型开始。如果所有工作良好, 优化过程中零的最佳数量的单位, 以适应扫描。</li>
		<li>请注意, 在运行优化过程时, "优化" 面板还会显示一个多色的进度条。</li>
		<li>在"绘图类型"框中使用不同的绘图选项,即轮廓图、误差 v N 单位、模型单位、累积振幅跟随优化过程。 注意: 优化过程中模型的变化是根据最新的模型不断更新的, 模型中的改进可以跟随不同的绘图选项和文本显示。<ol>
<li>等高线图<ol>
<li>利用等高线图, 通过模糊点并根据 x y 分布15的差异生成误差分数来评估模型的准确性。选择"比较"和"等高线图"显示, 以查看记录的和模型 CMAP 扫描之间的差异作为等高线图。正是这些差异, 优化过程尽量减少。 注意: "轮廓图" 使给定的刺激获得给定响应的概率密度。误差分数取决于原始和建模的等高线图之间的差异, 这可以通过在 "绘图" 面板中选择 "比较"选项和 "绘制类型" 面板中的 "轮廓图" 来可视化。红线表示原始扫描中的点密度更大, 绿线表示模型扫描中的更多点。顶部的差分图表明了最大误差的刺激强度。</li>
			</ol>
</li>
			<li>错误 v N 单位<ol>
<li>按照优化过程, 选择日志日志的错误 v N 单位的情节。优化过程的每个阶段都用不同的颜色绘制, 对应于"优化"面板中的条形。</li>
			</ol>
</li>
			<li>型号单位<ol>
<li>利用这一方法, 通过对模型中不同轴突的招聘, 来观察平均 CMAP 振幅是如何组成的;顶部显示了各电机单元的峰值振幅和阈值分布。</li>
			</ol>
</li>
			<li>累计振幅<ol>
<li>请注意, 这些单位的绘制顺序是增大大小, 而不是阈值。黑色曲线绘制累计单位数, 红色曲线显示累计振幅。</li>
			</ol>
</li>
		</ol>
</li>
	</ol>
</li>
	<li>优化过程完成后, 查看 "MScanFit 文本显示" 的分析结果。在文本显示上显示可用的 MScan 参数,即单位数、中值振幅、最大单位大小。</li>
	<li>单击"确定"按钮在 "保存适合内存文件" 框中保存模型在内存文件中进行进一步分析。</li>
</ol>4. CMAP 扫描免疫性使用。DAT 文件
注意: 软件的替代和免费版本可以对其他设备记录的 CMAP 扫描进行分析。
<ol><li>生成包含要分析的 CMAP 扫描的 a.DAT 文件。 注意: 这必须是一个标准的2列格式的文本文件, 在 mA 在一列的刺激强度和 CMAP 振幅在另一列。</li>
	<li>生成一个合适的。DAT 文件, 将两列从电子表格文件复制到文本编辑软件中, 然后将该扩展名指定为。DAT 而不是。Txt。<ol>
<li>执行 MScan 分析的所有其他步骤, 如前所述, 在启动独立程序并选择。DAT 文件。</li>
	</ol>
</li>
	<li>在伦敦大学学院的 FTP 站点上查找免费软件程序、手册和样本数据 (主机: 144.82.46.62, 用户名: QtracW, 密码: Hg32wK5e)。</li>
</ol>

<ol>
	<li>Sherrington, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=REMARKS+ON+THE+FOREGOING+LETTER.">REMARKS ON THE FOREGOING LETTER.</a> Can. Med. Assoc. J. 20, 66-67 (1929).</li><li>Gooch, C. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+unit+number+estimation:+a+technology+and+literature+review.">Motor unit number estimation: a technology and literature review.</a> Muscle Nerve. 50, 884-893 (2014).</li><li>McComas, A. J., Fawcett, P. R., Campbell, M. J., Sica, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+estimation+of+the+number+of+motor+units+within+a+human+muscle.">Electrophysiological estimation of the number of motor units within a human muscle.</a> J Neurol Neurosur Ps. 34, 121-131 (1971).</li><li>Oge, A. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+unit+number+estimation+in+transected+peripheral+nerves.">Motor unit number estimation in transected peripheral nerves.</a> Neurol Res. 32, 1072-1076 (2010).</li><li>Doherty, T. J., Brown, W. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+estimated+numbers+and+relative+sizes+of+thenar+motor+units+as+selected+by+multiple+point+stimulation+in+young+and+older+adults.">The estimated numbers and relative sizes of thenar motor units as selected by multiple point stimulation in young and older adults.</a> Muscle Nerve. 16, 355-366 (1993).</li><li>Bromberg, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+unit+estimation:+reproducibility+of+the+spike-triggered+averaging+technique+in+normal+and+ALS+subjects.">Motor unit estimation: reproducibility of the spike-triggered averaging technique in normal and ALS subjects.</a> Muscle Nerve. 16, 466-471 (1993).</li><li>Lomen-Hoerth, C., Slawnych, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Statistical+motor+unit+number+estimation:+from+theory+to+practice.">Statistical motor unit number estimation: from theory to practice.</a> Muscle Nerve. 28, 263-272 (2003).</li><li>Unlusoy Acar, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decline+of+compound+muscle+action+potentials+and+statistical+MUNEs+during+Wallerian+degeneration.">Decline of compound muscle action potentials and statistical MUNEs during Wallerian degeneration.</a> Neurophysiol. Clin. 44, 257-265 (2014).</li><li>Henderson, R. D., Ridall, P. G., Hutchinson, N. M., Pettitt, A. N., McCombe, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bayesian+statistical+MUNE+method.">Bayesian statistical MUNE method.</a> Muscle Nerve. 36, 206-213 (2007).</li><li>Ridall, P. G., Pettitt, A. N., Henderson, R. D., McCombe, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+unit+number+estimation--a+Bayesian+approach.">Motor unit number estimation--a Bayesian approach.</a> Biometrics. 62, 1235-1250 (2006).</li><li>Nandedkar, S. D., Nandedkar, D. S., Barkhaus, P. E., Stalberg, E. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+unit+number+index+(MUNIX).">Motor unit number index (MUNIX).</a> IEEE Trans. Biomed. Eng. 51, 2209-2211 (2004).</li><li>Neuwirth, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+Unit+Number+Index+(MUNIX):+a+novel+neurophysiological+marker+for+neuromuscular+disorders;+test-retest+reliability+in+healthy+volunteers.">Motor Unit Number Index (MUNIX): a novel neurophysiological marker for neuromuscular disorders; test-retest reliability in healthy volunteers.</a> Clin. Neurophysiol. 122, 1867-1872 (2011).</li><li>Jacobsen, A. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproducibility,+and+sensitivity+to+motor+unit+loss+in+amyotrophic+lateral+sclerosis,+of+a+novel+MUNE+method:+MScanFit+MUNE.">Reproducibility, and sensitivity to motor unit loss in amyotrophic lateral sclerosis, of a novel MUNE method: MScanFit MUNE.</a> Clin neurophysiol. 128, 1380-1388 (2017).</li><li>Maathuis, E. M., Drenthen, J., Visser, G. H., Blok, 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=Reproducibility+of+the+CMAP+scan.">Reproducibility of the CMAP scan.</a> J. Electromyogr. Kinesiol. 21, 433-437 (2011).</li><li>Sleutjes, B. T. H. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CMAP+scan+discontinuities:+automated+detection+and+relation+to+motor+unit+loss.">CMAP scan discontinuities: automated detection and relation to motor unit loss.</a> Clin Neurophysiol. 125, 388-395 (2014).</li><li>Bostock, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimating+motor+unit+numbers+from+a+CMAP+scan.">Estimating motor unit numbers from a CMAP scan.</a> Muscle Nerve. 53, 889-896 (2016).</li><li>DeLong, E. R., DeLong, D. M., Clarke-Pearson, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparing+the+areas+under+two+or+more+correlated+receiver+operating+characteristic+curves:+a+nonparametric+approach.">Comparing the areas under two or more correlated receiver operating characteristic curves: a nonparametric approach.</a> Biometrics. 44, 837-845 (1988).</li><li>Hanley, J. A., McNeil, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+of+comparing+the+areas+under+receiver+operating+characteristic+curves+derived+from+the+same+cases.">A method of comparing the areas under receiver operating characteristic curves derived from the same cases.</a> Radiology. 148, 839-843 (1983).</li><li>Farschtschi, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+action+potential+scans+and+ultrasound+imaging+in+neurofibromatosis+type+2+:+CMAP+Scans+and+Nerve+Imaging+in+NF2.">Muscle action potential scans and ultrasound imaging in neurofibromatosis type 2 : CMAP Scans and Nerve Imaging in NF2.</a> Muscle Nerve. 55, 350-358 (2017).</li></ol>

利益冲突: HB 从伦敦大学获得了他在本研究中使用的 Qtrac 软件的销售费。其他作者没有潜在的利益冲突。所有作者都批准了最后一篇文章。

议定书内的关键步骤:MScan 是一个高度自动化的程序, 但与所有的肌电信号的方法, 应注意取得一致的结果。在准备阶段, 重要的是要达到松弛, 因为自发活动或运动文物在 CMAP 扫描引入杂散方差在 CMAP 和混淆的世代的初步模型。
修改和疑难解答:我们发现, 手指的录音, 并使用听觉肌电反馈, 以帮助找到最佳肢体位置, 有助于避免一些差异。
<p class="jove...

马达系统运动依赖于马达单元, 它指的是单个的运动神经纤维和它激活的肌肉纤维, 而马达单位数是前角细胞或轴突支配单个肌肉1的数量。在神经支配和神经过程中, 健康的轴突接管了由侧枝发芽而失去的轴突的作用。因此, 复合肌肉动作电位 (CMAP) 振幅不提供有关电机单位损耗程度的必要信息。当超过50% 的马达单位丢失时, CMAP 振幅可能会开始下降。同样, 异常自发活动或运动单位电位 (内务部) 变化的幅度与神经支配程度无关。
总的来说, 没有电生理技术允许简单直接测量电机的单位数。相反, 对马达单位数 (免疫性) 的估计被用来评估较低的运动神经元损失2。自 1971年 McComas3引进的第一种方法, 即增量刺激免疫性的实施以来, 已开发出几种免疫性方法。大多数方法都是基于测量几种表面记录的电机单元电位 (sMUP), 并将最大 CMAP 除以平均 sMUP 振幅。这些方法包括增量刺激4, 多点刺激 (MPS)5和穗触发平均6。其他免疫性方法利用统计技术, 根据对刺激7,8,9,10的动力单元的发射概率性质。这种可变性意味着, 发射不同的马达单位组合会导致 CMAP 反应的大小变化。电机单元数指数 (Munix) 是一种较新近采用的方法, 它使用在自愿收缩期间记录的表面干扰模式来估计 sMUP11、12的平均大小。
这些免疫性方法都受到了一个或多个限制, 如主观性的存在, 对绝对 CMAP 振幅的依赖性, 单位选择中的偏倚, 取样足够长的时间, 或分析结果所需的长时间。最近开发了一种新的免疫性方法, 即 "CMAP 扫描免疫性" (MScan), 以克服这些限制13。这种方法避免了单位选择中固有的问题, 考虑到所有单位对 CMAP 的贡献, 在详细的刺激-反应曲线, 或 CMAP 扫描14,15测量。它还避免了类似的模型拟合方法9,10的扩展分析时间, 使用新的算法16。在最近的一项研究中, MScan 在估计运动单位数量方面的重现性优于两种传统方法, MPS 免疫性和 Munix13。此外, MScan 可能显示肌萎缩侧索硬化 (ALS) 早期阶段的运动单位损失比 MPS 免疫性和 Munix。MScan 比 MPS 免疫性快, Munix13。
本文详细介绍了 MScan 的方法。它还总结了以前报告的 MScan 在 ALS 和健康控制科目13患者内和评分员之间的重现性, 这可能使读者判断该方法是否适合于计划的研究。

<table>
	<tbody>
		<tr>
			<td>QtracW software</td>
			<td>Digitimer Ltd (copyright Institute of Neurology, University College, London)&#160;&#160;</td>
			<td>QtracW</td>
			<td></td>
		</tr>
		<tr>
			<td>MScanFit</td>
			<td>Digitimer Ltd (copyright Institute of Neurology, University College, London)&#160;&#160;</td>
			<td>QtracW</td>
			<td></td>
		</tr>
		<tr>
			<td>DS5 bipolar stimulator</td>
			<td>Digitimer Ltd&#160;</td>
			<td>DS5</td>
			<td></td>
		</tr>
		<tr>
			<td>D440 amplifier&#160;</td>
			<td>Digitimer Ltd&#160;</td>
			<td>D440-2 (2 channel) or D440-4 (4 channel)</td>
			<td></td>
		</tr>
		<tr>
			<td>HumBug Noise Eliminator</td>
			<td>Digitimer Ltd&#160;</td>
			<td>Humbug</td>
			<td></td>
		</tr>
		<tr>
			<td>Analogue-to-digital (A/D) board&#160;&#160;</td>
			<td>National Instruments</td>
			<td>NI-6221&#160;</td>
			<td></td>
		</tr>
	</tbody>
</table>

cmap scan mune (mscan) - a novel motor unit number estimation (mune) method

与其他电机单位数估计方法 (免疫性) 一样, 复合肌肉动作电位 (CMAP) 扫描免疫性 (MScan) 是一种非侵入性电生理学方法来估计肌肉中运转的运动单位数。免疫性是评价神经病和 neuronopathies 的重要工具。不同于使用的大多数免疫性方法, MScan 评估肌肉中的所有马达单位, 通过拟合模型到详细的刺激反应曲线, 或 CMAP 扫描。它从而避免了所有免疫性方法的固有偏差, 其基础是从一个小样本单位推断出来的。像 ' 贝叶斯免疫性, ' MScan 分析工作的模型, 由不同的振幅, 阈值和阈值多变性的电机单位, 但拟合方法是相当不同的, 并完成五分钟内, 而不是几个小时。MScan 离线分析工作分为两个阶段: 一是基于扫描点的斜率和方差产生的初始模型, 其次, 通过调整所有参数来改进该模型, 以提高原始扫描与由模型生成的扫描。
这一新的方法已被测试的重现性和记录时间22肌萎缩侧索硬化症 (ALS) 患者和20健康的控制, 每一次测试重复两次双盲医生。MScan 显示优异的内部和评分的重复性与 ICC 的价值 > 0.98 和变化系数平均 12.3 1.6%。两名观察员的评分员内重现性没有差异。平均记录时间为 6.27 0.27 分钟。
本协议描述如何记录 CMAP 扫描以及如何使用 MScan 软件来得出对运行的马达单元的数量和大小的估计。MScan 是一种快速、方便、重现性好的方法, 有助于诊断和监测神经肌肉疾病的进展。

在最近的一项研究中获得了以下结果: MScan 法与两种已建立的技术进行了比较, 即多点刺激免疫性 (MPS) 和电机单位数指数 (Munix)13。结果表明, 本协议所描述的技术具有良好的重现性, 可实现一致的结果。该方法能使 ALS 患者比 CMAP 早期的健康控制更加容易, 使用速度快,13。这表明该方法可能适合临床使用。
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Watch this Scientific Journal Video about CMAP Scan MUNE MScan - A Novel Motor Unit Number Estimation MUNE Method at JoVE.com

CMAP Scan MUNE MScan - A Novel Motor Unit Number Estimation MUNE Method

该协议描述了一种新的方法来估计肌肉中运转的运动单位的数量, 通过拟合一个模型到一个详细的刺激反应曲线的复合肌肉动作电位。它快速, 易于执行和分析, 具有优良的重现性。

CMAP 扫描免疫性 (MScan)-一种新的电机单元数估计 (免疫性) 方法

Like other methods for motor unit number estimation (MUNE), compound muscle action potential (CMAP) scan MUNE (MScan) is a non-invasive electrophysiologic ...

cmap-scan-mune-mscan-a-novel-motor-unit-number-estimation-mune-method

Research

JoVE Journal

Neuroscience

CMAP Scan MUNE (MScan) - A Novel Motor Unit Number Estimation (MUNE) Method

12.2K 次观看.  Aarhus University Hospital. 该协议描述了一种新的方法来估计肌肉中运转的运动单位的数量, 通过拟合一个模型到一个详细的刺激反应曲线的复合肌肉动作电位。它快速, 易于执行和分析, 具有优良的重现性。

视频: CMAP Scan MUNE MScan - A Novel Motor Unit Number Estimation MUNE Method

Novel Apparatus and Method for Drug Reinforcement

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction. Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in animal research to model various components of drug reinforcement, consumption, and addiction in humans. For this study, we used a novel approach to studying drug reinforcement in rats by combining traditional CPP and self-administration methodologies. We assembled an apparatus using two Med Associate operant chambers, sensory stimuli, and a Plexiglas-constructed neutral zone. These modifications allowed our experiments to encompass motivational aspects of drug intake through self-administration and drug-free assessment of drug/cue conditioning strength with the CPP test. In our experiments, rats self-administered cocaine (0.75 mg/kg/inj, i.v.) during either four (e.g., the "short-term") or eight (e.g., the "long-term") alternating-day sessions in an operant environment containing distinctive sensory cues (e.g., olfactory and visual). On the alternate days, in the other (differently-cued) operant environment, saline was available for self-infusion (0.1 ml, i.v.). Twenty-four hours after the last self-administration/cue-pairing session, a CPP test was conducted. Consistent with typical CPP findings, there was a significant preference for the chamber associated with cocaine self-administration. In addition, in animals undergoing the long-term experiment, a significant positive correlation between CPP magnitude and the number of cocaine-reinforced lever responses. In conclusion, this apparatus and approach is time and cost effective, can be used to examine a wide array of topics pertaining to drug abuse, and provides more flexibility in experimental design than CPP or self-administration methods alone.

Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in research to model various components of drug reinforcement, consumption, and addiction in humans. In this report, we combined traditional CPP and self-administration methods as a novel approach to studying drug reinforcement and addiction in rats.

新型仪器和药物加固方法

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction.  Operant drug ...

科研

神经科学

Presynaptic Dopamine Dynamics in Striatal Brain Slices with Fast-scan Cyclic Voltammetry

Extensive research has focused on the neurotransmitter dopamine because of its importance in the mechanism of action of drugs of abuse (e.g. cocaine and amphetamine), the role it plays in psychiatric illnesses (e.g. schizophrenia and Attention Deficit Hyperactivity Disorder), and its involvement in degenerative disorders like Parkinson's and Huntington's disease. Under normal physiological conditions, dopamine is known to regulate locomotor activity, cognition, learning, emotional affect, and neuroendocrine hormone secretion. One of the largest densities of dopamine neurons is within the striatum, which can be divided in two distinct neuroanatomical regions known as the nucleus accumbens and the caudate-putamen. The objective is to illustrate a general protocol for slice fast-scan cyclic voltammetry (FSCV) within the mouse striatum. FSCV is a well-defined electrochemical technique providing the opportunity to measure dopamine release and uptake in real time in discrete brain regions. Carbon fiber microelectrodes (diameter of ~ 7 &mu;m) are used in FSCV to detect dopamine oxidation. The analytical advantage of using FSCV to detect dopamine is its enhanced temporal resolution of 100 milliseconds and spatial resolution of less than ten microns, providing complementary information to in vivo microdialysis.

Using fast-scan cyclic voltammetry to measure electrically evoked presynaptic dopamine dynamics in striatal brain slices.

突触前多巴胺纹状体脑片的快速扫描循环伏安动态

Extensive research has focused on the neurotransmitter dopamine because of its importance in the mechanism of action of drugs of abuse (e.g. cocaine and ...

A Functional Motor Unit in the Culture Dish: Co-culture of Spinal Cord Explants and Muscle Cells

Human primary muscle cells cultured aneurally in monolayer rarely contract spontaneously because, in the absence of a nerve component, cell differentiation is limited and motor neuron stimulation is missing1. These limitations hamper the in vitro study of many neuromuscular diseases in cultured muscle cells. Importantly, the experimental constraints of monolayered, cultured muscle cells can be overcome by functional innervation of myofibers with spinal cord explants in co-cultures.
Here, we show the different steps required to achieve an efficient, proper innervation of human primary muscle cells, leading to complete differentiation and fiber contraction according to the method developed by Askanas2. To do so, muscle cells are co-cultured with spinal cord explants of rat embryos at ED 13.5, with the dorsal root ganglia still attached to the spinal cord slices. After a few days, the muscle fibers start to contract and eventually become cross-striated through innervation by functional neurites projecting from the spinal cord explants that connecting to the muscle cells. This structure can be maintained for many months, simply by regular exchange of the culture medium.
The applications of this invaluable tool are numerous, as it represents a functional model for multidisciplinary analyses of human muscle development and innervation. In fact, a complete de novo neuromuscular junction installation occurs in a culture dish, allowing an easy measurement of many parameters at each step, in a fundamental and physiological context.
Just to cite a few examples, genomic and/or proteomic studies can be performed directly on the co-cultures. Furthermore, pre- and post-synaptic effects can be specifically and separately assessed at the neuromuscular junction, because both components come from different species, rat and human, respectively. The nerve-muscle co-culture can also be performed with human muscle cells isolated from patients suffering from muscle or neuromuscular diseases3, and thus can be used as a screening tool for candidate drugs. Finally, no special equipment but a regular BSL2 facility is needed to reproduce a functional motor unit in a culture dish. This method thus is valuable for both the muscle as well as the neuromuscular research communities for physiological and mechanistic studies of neuromuscular function, in a normal and disease context.

Cultured muscle cells are an inadequate model to recapitulate innervated muscle in vivo. A functional motor unit can be reproduced in vitro by innervation of differentiated human primary muscle cells using rat embryo spinal cord explants. This article describes how co-cultures of spinal cord explants and muscle cells are established.

一个在培养皿中的功能电机组:脊髓植和肌肉细胞共培养

Human primary muscle cells cultured  aneurally in monolayer rarely contract spontaneously because, in the absence of  a nerve component, cell ...

Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In vivo

The spinal motoneuron has long been a good model system for studying neural function because it is a neuron of the central nervous system with the unique properties of (1) having readily identifiable targets (the muscle fibers) and therefore having a very well-known function (to control muscle contraction); (2) being the convergent target of many spinal and descending networks, hence the name of "final common pathway"; and (3) having a large soma which makes it possible to penetrate them with sharp intracellular electrodes. Furthermore, when studied in vivo, it is possible to record simultaneously the electrical activity of the motoneurons and the force developed by their muscle targets. Performing intracellular recordings of motoneurons in vivo therefore put the experimentalist in the unique position of being able to study, at the same time, all the compartments of the "motor unit" (the name given to the motoneuron, its axon, and the muscle fibers it innervates1): the inputs impinging on the motoneuron, the electrophysiological properties of the motoneuron, and the impact of these properties on the physiological function of the motoneurons, i.e. the force produced by its motor unit. However, this approach is very challenging because the preparation cannot be paralyzed and thus the mechanical stability for the intracellular recording is reduced. Thus, this kind of experiments has only been achieved in cats and in rats. However, the study of spinal motor systems could make a formidable leap if it was possible to perform similar experiments in normal and genetically modified mice.
For technical reasons, the study of the spinal networks in mice has mostly been limited to neonatal in vitro preparations, where the motoneurons and the spinal networks are immature, the motoneurons are separated from their targets, and when studied in slices, the motoneurons are separated from most of their inputs. Until recently, only a few groups had managed to perform intracellular recordings of motoneurons in vivo2-4 , including our team who published a new preparation which allowed us to obtain very stable recordings of motoneurons in vivo in adult mice5,6. However, these recordings were obtained in paralyzed animals, i.e. without the possibility to record the force output of these motoneurons. Here we present an extension of this original preparation in which we were able to obtain simultaneous recordings of the electrophysiological properties of the motoneurons and of the force developed by their motor unit. This is an important achievement, as it allows us to identify the different types of motoneurons based on their force profile, and thereby revealing their function. Coupled with genetic models disturbing spinal segmental circuitry7-9, or reproducting human disease10,11, we expect this technique to be an essential tool for the study of spinal motor system.

Simultaneous Intracellular Recording of a Lumbar Motoneuron and the Force Produced by its Motor Unit in the Adult Mouse In vivo

This new method permits the simultaneous intracellular recording of a single adult mouse motoneuron and the measurement of the force produced by its muscle fibers. The combined investigation of the electrical and mechanical properties of motor units in normal and genetically modified animals is a breakthrough for the study of the neuromuscular system.

同时细胞内记录的车组在成年小鼠的腰椎运动神经元产生的力<em在体内</em

The  spinal motoneuron has long been a good model system for studying neural  function because it is a neuron of the central nervous system with the ...

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica

In animals with large identified neurons (e.g. mollusks), analysis of motor pools is done using intracellular techniques1,2,3,4. Recently, we developed a technique to extracellularly stimulate and record individual neurons in Aplysia californica5. We now describe a protocol for using this technique to uniquely identify and characterize motor neurons within a motor pool.
This extracellular technique has advantages. First, extracellular electrodes can stimulate and record neurons through the sheath5, so it does not need to be removed. Thus, neurons will be healthier in extracellular experiments than in intracellular ones. Second, if ganglia are rotated by appropriate pinning of the sheath, extracellular electrodes can access neurons on both sides of the ganglion, which makes it easier and more efficient to identify multiple neurons in the same preparation. Third, extracellular electrodes do not need to penetrate cells, and thus can be easily moved back and forth among neurons, causing less damage to them. This is especially useful when one tries to record multiple neurons during repeating motor patterns that may only persist for minutes. Fourth, extracellular electrodes are more flexible than intracellular ones during muscle movements. Intracellular electrodes may pull out and damage neurons during muscle contractions. In contrast, since extracellular electrodes are gently pressed onto the sheath above neurons, they usually stay above the same neuron during muscle contractions, and thus can be used in more intact preparations.
To uniquely identify motor neurons for a motor pool (in particular, the I1/I3 muscle in Aplysia) using extracellular electrodes, one can use features that do not require intracellular measurements as criteria: soma size and location, axonal projection, and muscle innervation4,6,7. For the particular motor pool used to illustrate the technique, we recorded from buccal nerves 2 and 3 to measure axonal projections, and measured the contraction forces of the I1/I3 muscle to determine the pattern of muscle innervation for the individual motor neurons.
We demonstrate the complete process of first identifying motor neurons using muscle innervation, then characterizing their timing during motor patterns, creating a simplified diagnostic method for rapid identification. The simplified and more rapid diagnostic method is superior for more intact preparations, e.g. in the suspended buccal mass preparation8 or in vivo9. This process can also be applied in other motor pools10,11,12 in Aplysia or in other animal systems2,3,13,14.

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica

In animals with large identified neurons (e.g. mollusks), analysis of motor pools is done using intracellular techniques1,2,3,4. Recently, we developed a technique to extracellularly stimulate and record individual neurons in Aplysia californica5. We now describe a protocol for using this technique to uniquely identify and characterize motor neurons within a motor pool.

运动神经元细胞外识别的肌肉运动游泳池<em&gt;海兔夜蛾</em

In animals with large  identified neurons (e.g. mollusks), analysis of motor pools is done using  intracellular techniques1,2,3,4. Recently,  we developed ...

A Novel Method of Drug Administration to Multiple Zebrafish (Danio rerio) and the Quantification of Withdrawal

Anxiety testing in zebrafish is often studied in combination with the application of pharmacological substances. In these studies, fish are routinely netted and transported between home aquaria and dosing tanks. In order to enhance the ease of compound administration, a novel method for transferring fish between tanks for drug administration was developed. Inserts that are designed for spawning were used to transfer groups of fish into the drug solution, allowing accurate dosing of all fish in the group. This increases the precision and efficiency of dosing, which becomes very important in long schedules of repeated drug administration. We implemented this procedure for use in a study examining the behavior of zebrafish in the light/dark test after administering ethanol with differing 21 day schedules. In fish exposed to daily-moderate amounts of alcohol there was a significant difference in location preference after 2 days of withdrawal when compared to the control group. However, a significant difference in location preference in a group exposed to weekly-binge administration was not observed. 
This protocol can be generalized for use with all types of compounds that are water-soluble and may be used in any situation when the behavior of fish during or after long schedules of drug administration is being examined. The light/dark test is also a valuable method of assessing withdrawal-induced changes in anxiety.

A Novel Method of Drug Administration to Multiple Zebrafish Danio rerio and the Quantification of Withdrawal

A novel method for reducing variability when exposing fish to drugs is explained. Fish exposed to various patterns of ethanol exposure were found to have altered anxiety levels during withdrawal in a light/dark scototaxic assay.

药品监督管理局多个斑马鱼的新方法（<em&gt;斑马鱼</em&gt;）和退出的量化

Anxiety testing in zebrafish is often studied in combination with the application of pharmacological substances. In these studies, fish are routinely ...

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

使用新颖的方法在自由移动的新生大鼠中记录脑电图

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

Chronic Traumatic Encephalopathy (CTE) is an established neurodegenerative disease that is closely associated with exposure to repetitive mild Traumatic Brain Injury (mTBI). The mechanisms responsible for its complex pathological changes remain largely elusive, despite a recent consensus to define the neuropathological criteria. Here, we describe a novel method to develop a model of CTE in Drosophila melanogaster&#160;(Drosophila&#160;)&#160;in an attempt to identify the key genes and pathways that lead to the characteristic hyperphosphorylated tau accumulation and neuronal death in the brain. Adjustable-strength impacts to inflict mild closed injury are delivered directly to the fly head, subjecting the head to rapid acceleration and deceleration. Our method eliminates the potential problems inherent with other Drosophila mTBI models (e.g.,animal death might be induced by damage to other parts of the body or to internal organs). The less labor- and cost-intensive animal care, short life span, and extensive genetic tools make the fruit fly ideal to study CTE pathogenesis and make it possible to perform large-scale, genome-wide forward genetic and pharmacological screens. We anticipate that the ongoing characterization of the model will generate important mechanistic insights on disease prevention and therapeutic approaches.

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

Here, we describe a new approach to inflict closed-head traumatic brain injury in Drosophila melanogaster. Our method has the advantage of directly delivering repetitive impacts with adjustable strength to the head alone. Further exploration of the invertebrate system will help to illuminate the pathogenesis of chronic traumatic encephalopathy.

慢性创伤性脑病模型的一种新方法<em&gt;果蝇</em

Chronic Traumatic Encephalopathy (CTE) is an established neurodegenerative disease that is closely associated with exposure to repetitive mild Traumatic ...

Stereological Estimation of Dopaminergic Neuron Number in the Mouse Substantia Nigra Using the Optical Fractionator and Standard Microscopy Equipment

In pre-clinical Parkinson&#39;s disease research, analysis of the nigrostriatal tract, including quantification of dopaminergic neuron loss within the substantia nigra, is essential. To estimate the total dopaminergic neuron number, unbiased stereology using the optical fractionator method is currently considered the gold standard. Because the theory behind the optical fractionator method is complex and because stereology is difficult to achieve without specialized equipment, several commercially available complete stereology systems that include the necessary software do exist, purely for cell counting reasons. Since purchasing a specialized stereology setup is not always feasible, for many reasons, this report describes a method for the stereological estimation of dopaminergic neuronal cell counts using standard microscopy equipment, including a light microscope, a motorized object table (x, y, z plane) with imaging software, and a computer for analysis. A step-by-step explanation is given on how to perform stereological quantification using the optical fractionator method, and pre-programmed files for the calculation of estimated cell counts are provided. To assess the accuracy of this method, a comparison to data obtained from a commercially available stereology apparatus was performed. Comparable cell numbers were found using this protocol and the stereology device, thus demonstrating the precision of this protocol for unbiased stereology.

This work presents a step-by-step protocol for the unbiased stereological estimation of dopaminergic neuronal cell numbers in the mouse substantia nigra using standard microscopy equipment (i.e., a light microscope, a motorized object table (x, y, z plane), and public domain software for digital image analysis.

用光学分馏塔和标准显微镜设备对小鼠黑质多巴胺能神经元数的视学估计

In pre-clinical Parkinson&#39;s disease research, analysis of the nigrostriatal tract, including quantification of dopaminergic neuron loss within the ...

Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets

In vitro autoradiography aims to visualize the anatomical distribution of a protein of interest in tissue from experimental animals as well as humans. The method is based on the specific binding of a radioligand to its biological target. Therefore, frozen tissue sections are incubated with radioligand solution, and the binding to the target is subsequently localized by the detection of radioactive decay, for example, by using photosensitive film or phosphor imaging plates. Resulting digital autoradiograms display remarkable spatial resolution, which enables quantification and localization of radioligand binding in distinct anatomical structures. Moreover, quantification allows for the pharmacological characterization of ligand affinity by means of dissociation constants (Kd), inhibition constants (Ki) as well as the density of binding sites (Bmax) in selected tissues. Thus, the method provides information about both target localization and ligand selectivity. Here, the technique is exemplified with autoradiographic characterization of the high-affinity &#947;-hydroxybutyric acid (GHB) binding sites in mammalian brain tissue, with special emphasis on methodological considerations regarding the binding assay parameters, the choice of the radioligand and the detection method.

The method of autoradiography is routinely used to study binding of radioligands to tissue sections for determination of qualitative or quantitative pharmacology.

自体影像学作为一种简单而有力的药理靶标可视化和表征方法

In vitro autoradiography aims to visualize the anatomical distribution of a protein of interest in tissue from experimental animals as well as humans. The ...