Name: automated analysis of c. elegans swim behavior using celest software
Uploaded: 2016-12-07T12:30:00.000+00:00
Duration: 8 min 47 s
Description: Watch this Scientific Journal Video about Automated Analysis of C. elegans Swim Behavior Using CeleST Software at JoVE.com

张玉华发展由美国国立卫生研究院的支持授予R21AG027513和U01AG045864。数据和一些简短的视频表示是改编自Restif 等。 （2014年）8，这是根据知识共享署名（CC BY）牌照http://creativecommons.org/licenses/by/4.0/出版。我们感谢里卡多Laranjeiro稿件帮助。

1.线虫生长与处理<ol><li> C.生长在含NGM发现与OP50-1 大肠杆菌作为食物来源1-3标准培养皿线虫 。 </li><li>保持培养物中维持所需温度的恒温箱。 线虫生长良好，从15至25℃，以20℃为标准生长温度。 注:游泳是一个强大的表型，但游泳的活力可能是由于环境因素，包括在平板上的污染和过热的文化和幻灯片的影响。强努力保持稳定的条件应该作出。 </li><li>使用由铂丝阻燃密封入玻璃棒处理单动物4转移的一个自制的线虫挑。 </li></ol> 2.设置游泳分析<ol><li>使用由立体显微镜，数码CCD摄像头和数字视频录制软件的集成系统。天书软件CAN为一个单独的计算机上使用。 <ol><li>盖上暗布的显微镜 （ 如毡），以防止眩光在显微镜载玻片游泳降面积，提高图像质量。 </li><li>调整显微镜的工作距离和放大倍率的游泳区的全视图，基镜获得的映衬下动物的鲜明对比。将照明到暗视野可视化的成年人的身体作为白色对黑色的背景，这非常适用于计算可视化和与游泳分析软件跟踪。 </li><li>注意保持工作区域，显微镜载玻片，和游泳降得很干净。杂物可通过堵塞成年人的自由游动的干扰分析，和纤维的细丝可以作为跟踪过程中不运动的动物进行检测。 </li></ol></li></ol> 3.动物的制备为液体体质分析<ol><li>吸取60μLØ˚F1X M9缓冲区中预先打印载玻片10毫米环。确保滴完全覆盖环的内侧区域。该软件提取使用包含在幻灯片上会比利用，如果多孔板这个墨滴大小的更多信息，游泳。 </li><li>接单成人线虫，并将它们传送到1×M9缓冲液中滴，最大限度地减少细菌，可以云中的液体的运输。首先将动物转移到不包含很多细菌草坪上板的一侧，让他们爬行，然后轻轻提起他们，尽量减少细菌。 <ol><li>当线虫转移到下降，用接近的分析站解剖范围，以减少的游泳时间的长度，这可能会影响动物25的行为。 </li><li>将只有4个成年人在下降，如果他们大力移动和交叉路径频繁。重要思想为指导，以减少重叠的游泳者（这混淆了计算分析），考虑将4只动物每一滴水的时候都那么积极，年轻，健康成人WT每下降5只动物，当他们年长显示较少的活力。 注:健康WT线虫本能地开始被放置在液体后不久游泳。随着线虫挑的帮助下，轻轻地大人在下降，如果他们聚集分开。 </li></ol></li></ol> 4.体质分析 - 液 - 游泳录像<ol><li>装有显微镜来记录他们的游泳的基础上成人的滑动位置。 30秒在液体中的个体动物的行为的视频捕获细节（ 视频1），以及大量的这种记录时由CELEST软件8,26计算机辅助是很容易实现的采样;然而，较长的时间游泳可能是理想的其他特定行为研究25。重要的是，一定要保持整个所有记录相同的放大倍率的放大倍率，因为不同affecŧ游泳分析和意志偏差比较。 注:提供与使用的材料表中列出的数字视频录制软件的指导，这些步骤; VirtualDub的开放式访问软件可以替代，虽然我们还没有测试过这一点。 <ol><li>使用以下设置作为成功的游泳分析指导:696点¯x520像素的图像尺寸，0.02毫米/像素的图像分辨率和18帧/ s的速率。这些设置更高分辨率的版本可能有利于跟踪，但不会影响程序计算的措施。 注:1倍M9的缓冲区上的显微镜载玻片60μL下降所提供的游泳区没有太多的深度，这最大限度地减少向上和向下运动游泳。 </li></ol></li><li>在设置下，进入"录音"，载"限价序列大小"到"磁盘上的545架"。在相同的标题下，选中"使用录像管理器"对话框。点击"修改当前的录音功丁脚本..."按钮，然后在"命令"，选择"抓取窗格"，并根据"停止条件"，输入"停止后，0:00:30.00"。 </li><li>要录制，选择家庭标题下的"在RAM新建序列"，然后按下红色的"录制"按钮。该软件将在抢30秒545帧，数码相机和保持他们的RAM内存。 </li></ol> 5.体质液体分析 - 视频处理<ol><li>通过点击标题的家，并在视频录制软件，选择"导出全序列为...&#39;片段视频为图像帧序列。 注:数字视频录制软件可以保存帧作为.BMP，.JPG，.TIF或.png，所有这一切都可以通过这里介绍的游泳分析程序读取。需要注意的是.TIF文件使用更多的存储空间，但一些格式如.jpg可能会降低图像质量。 </li><li>启动从MATLAB B中的游泳分析程序Ÿ点击运行按钮。 </li><li>在游泳分析程序的主屏幕上，使用"添加一个视频"或"添加多个视频"按钮来上传图片序列帧一次或同时附上相关的标签，如基因型，日期，审判和持续时间组。该软件创建标记序列的数据库。 </li><li>通过点击&#39;1启用上传序列多个动物的同步跟踪。过程的视频"。在新的屏幕中，选择在左边框中输入所需的视频，然后单击"添加到列表"。对于每一个视频，在右上角的列表中单击其名称右下角显示视频的第一张图像。通过选择环内3分划定游泳区，并通过单击"进程上面列出的所有视频"启动处理。查看在中心底部的处理的进展情况。 </li><li>一旦完成，"关闭"视频处理画面。 </li></ol> 6。体质液体分析 - 测量<ol><li>点击"2。计算措施&#39;，打开一个新的屏幕，其中所处理的视频可以上传，一次一个，以验证或拒绝单个动物的跟踪。已处理的视频会出现在在哪里可以点击视频名称上传该视频左侧的列表中。 </li><li>使用本手册一步把重点放在对比成功跟踪（绿色街区）错过了跟踪（红色块）区域。 注意:可替换地，人们可以依靠有效跟踪的自动输出，因为该软件具有（如在2020动物中观察到从404影片8 94.1％效力）一个鲁棒跟踪系统。 <ol><li>找到"分段体的有效性"中的下半部分在屏幕上的第一个颜色条。 "有效框架"的比例为绿色，而"拒绝框架"的比例为红色。有效帧的≥80％意味着该跟踪我载文信息用于计算措施来自于一个30秒至少432帧出540游泳记录在18帧/秒的审判。 </li><li>轻松浏览和修改的有效块/拒绝使用"下一步块"，"开关有效性"，"分割块"和"隔离框"选项框。在极少数情况下2只动物在拥挤的游泳区交叉路径，切换其身份证号码在特定帧范围的"开关"按钮。 </li><li>查看在屏幕的下半部分的条形图上的跟踪和重叠的表演的附加信息。分割体（每帧动物的体长）的长度重叠发生率相关。 </li></ol></li><li>如果该视频的任何亮点干扰分析，通过选择"眩光区"功能调整的游泳区明亮区域。 </li><li>点击"保存并计算措施"保存有效期输入之前最多加载另一个处理的视频。在这个阶段，软件计算，在后台（未示出给用户），动物个体的游泳性能的曲率图和如下参数（ 视频2）。 注:如果集中在这里测得的参数在表1中描述。波引发速率，体波数，不对称，拉伸和卷曲基于该软件计算用于动物的曲率图计算出的;行进速度，笔触和活动指数参数不是基于曲率的地图。 </li><li>点击"关闭"退出"计算的措施"屏幕。 </li></ol> 7.体质分析 - 液 - 结果输出注:该软件可以在游泳运动的多种措施，从公开到很细微的行为特征用肉眼不容易打进封面报道（ 视频3和4）。在这里，重点是8个参数，通常表现出良好的动态范围:波引发速率，身体波数，不对称，拉伸，卷边，行驶速度，笔触和活动指数。 <ol><li>在程序的主界面，点击"3。显示结果"来获取分析输出。 </li><li>在左边的框中选择想要的画面序列进行分析，并将其移动到新的或现有的样本组通过单击"添加到所选样本"或"添加到新样品"按钮创建进行统计学处理样本组。 </li><li>点击"图形显示这些样品"去，通过选择显示在屏幕顶部的曲线图和样品为每8个参数的关键统计数据的第二个屏幕。 <ol><li>如果需要的话，使用"颜色"，"直方图＃&#39;，和选择"蠕虫＃&#39;调整条形图的表示在屏幕的左上角。 </li></ol></li><li>使用"2D直方图&#39;通过与"测量在Y轴上"下拉在屏幕的左边的菜单的"在X轴上测度绘制任何两个参数的组合。单击"关闭"退出并返回到"显示结果"屏幕。 </li><li>使用屏幕左上方的"导出..."按钮来详细的数值数据保存为.csv文件打开并在电子表格程序进一步操纵。 </li><li> &#39;关闭&#39;的&#39;显示结果&#39;屏幕回到主屏幕，然后单击"退出"，以确保包含分析视频序列数据库的保存。 </li></ol>

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The authors have nothing to disclose.

作为模型系统的使用线虫的继续，由于其遗传延展性，实验易处理性和解剖学注释以细致详细地增加。例如， 线虫雌雄同体的神经元结构和连接清楚地映射出34-36，极大地方便了控制特定行为的特定神经元回路的调查。 302的神经元构成了成人雌雄同体的神经系统，其处理范围广泛的感觉输入到像运动基本行为输出。更复杂的男性的神经系统的结构也被描述37，使性别特异性电路加以处理。 37 线虫行为已被广泛地研究了含有固体培养基的标准培养板中。由于WT 线虫在可预见的正弦波琼脂填充板从移动，偏差整体格局可以通过眼睛进行检测和人工评分。这种方法，然而，是受实验者的标准，是劳动密集型的。硬件和软件工具，用于跟踪和测量线虫在固体培养基上运动删除的主观性偏差和允许大规模的研究，允许更多复杂的生物学问题加以解决。最近由谢弗实验室16创建的行为数据库的分析扩展和深度已与上板运动的计算系统来实现的一个很好的例子。 当WT 线虫放入液体，动物很快适应了新的环境运动，开始游泳。 线虫游泳利用运动的更大的范围比爬行和可以更不规则8。像张玉华软件旨在填补C.详细分析差距线虫液体，permi行为的那些不容易通过的未受助眼测定，或可以比手动得分更迅速地完成运动相关的参数拟合定量。在8硬H成个人可以处理多达200影片，1000〜记录每天。 软件定义游泳充当体能和行为进行了全面评估指纹参数。除了富集在液体线虫行为和它们的基础的分子途径的复杂的小面的理解，该软件可用于探索生物学的多个方面，包括药理学应答，老化，和不同的行为。这里介绍的，那发生在C的物理性能的量化变化概述线虫成年人随着年龄的增长是该软件的这种应用的一个例子（更详细的介绍，请参阅Restif 等8）。在老龄化的背景下，一些测量参数下降，而更为增加或没有在野生型一致地变化。趋势是由长寿突变体的计算轮廓证实，在很大程度上，和同年龄人群的优雅和坏几岁同伙的相对配置文件保存在统一的环境条件。该软件的高分辨率也可以揭示细微的表现型先前在广泛表征突变体（ 例如， 年龄-1（hx546）在图1中）未知。 有描述的协议的几个特别关键的步骤。维护游泳环境应变片文化的一个恒定的温度环境，是游泳重复性重要的，所以实验者我们非常鼓励去煞费苦心，避免随意的温度变化。游泳媒体应在相同的温度下将板。同样地，仔细注意墨滴大小为游泳将有助于确保重现性。最后，谨慎的做法是在思考推进有关积累大量的视频文件的卸载。建议处理从视频采集计算机除了一个网站的图片。 利用这里提出供游泳分析软件的具有一定的局限性。首先，虽然程序可以同时跟踪多个动物，如果超过五只动物被联合分析，有一个增加的机会，该动物将在视频图像跨越彼此游泳。当程序不能明确地确定哪些动物是它，它审查这些数据帧。虽然此方案功能确保个别动物数据是高质量的，它限制了吞吐量。其次，图像应该还算干净，就是无灰尘，污迹和灯光刺眼，因为相关的信号也可以混淆图像分析。作为协议第2.1.1节，一个非常低的高科技投入，可以通过消除并发症的波动大大有助于图像捕捉注意到我Ñ环境照明是覆盖用深色布不允许环境光到达阶段舞台区。第三，该程序用于在成年阶段的动物进行了优化。低龄幼虫游泳速度非常快，有小的机构，这增加了程序错误。第四，一些软件利用MATLAB中，并且当存在版本升级和/或操作系统升级，一些程序链接可能会中断。目前，该软件对MATLAB 2015B和Mac OS版10.10进行了优化，但我们预计很快就会发布一个软件版本，反对这种变化更加健壮。最后，视频数据文件可以变得很大快，并且需要的存储空间被分配。 总之，这里介绍的是一种简单的方法，可以通过任何实验室容易地实现没有太多投资创建线虫游泳CELEST分析的影片。软件包的功能包括广泛的自动化，从通过跟踪分析，同时多动物跟踪，以及使用的数学基础（ 即，曲率措施）对于大多数运动参数的定量。该软件是开源的，其代码和演示公开可作为Restif 等详细说明。 8。虽然该方案特点进行跟踪先进的计算机视觉分析，其他已发布跟踪系统（ 例如 ，Greenblum 等人 ，2014年38），与这里介绍的软件的参数分析兼容。未来的改进将朝向的软件转换成一个更强大的包，不限制使用上面提到的操作系统（在材料的表也表示）的特定版本被引导。

定义遗传学，表观遗传学，经验和环境的影响的行为是如何在现代生物学的一个重大挑战。操作简单，适合遗传模型，可以通过计算跟踪和测量可以实现这一目标作出重要贡献。该线虫就是这样的一个模式。本文的目的是演示线虫游泳运动如何可以跟踪和定量提取八与生物相关性新颖特征的信息。 线虫具有约2相对短的寿命- 3瓦特和大约4天在20℃ 的1,2-生殖周期。在标准的实验室培养，这种微小线虫生长在含线虫生长介质（NGM）培养皿正在流传着一种细菌的食物来源3,4。 WT N2动物琼脂填充板优雅正弦波积极动;他们改变移动鼠漫游时（食品求），住宅（浏览），或从餐（无效饱腹感静止期）回收ES 5。减值6或7-12岁还可以显着地改变上板的运动。 基因调节行为，长期保健或老化，可以功能特点，通过分析线虫的运动模式或运动。衡量长期保健方法之一是到老年人分为根据自己的平板上运动三大类（ 如 A，B和C），剧烈体力活动是A类和麻痹是C类7,8,13。虽然这样的定性排序是揭示体质的差异是有用的，分类是没有明确的界限笼统，因而他们的得分是受实验者的偏见。 越来越多的纤夫有升高的客观性，复杂性，和C的分析精度线虫运动在固体培养基上14-18。 线虫在平板上运动主要是受限制的，其中该动物使得与媒体的固体表面接触的平面。 （ 线虫也可以通过从身体仍然与固体表面接触的其余部分的抬起头离开，定位体在多个平面诱导探索。然而，这种行为是不寻常的。）当放置在液体中，这线虫发起波动的运动，或游泳，比在固体表面的动物，具有更广泛的空间运动，速度和运动的深度更大范围，并与前从后移动不同年龄的发病率增加。由于体能和应对新环境的快速分析，实验者可以将动物个体成液滴和得分它的身体弯曲频率下的解剖范围。录像可以方便的游泳活力的现在和未来的刻痕动物。然而，手动方法限制了可被打进特征的数目，并完全约束在一次得分一只动物。 运动在液体比固体培养基上运动少了探讨。实际上，有一些是健壮和容易在实验室中实施以测量液体19-24运动几个软件选项。该软件张玉华（ 线虫游泳测试）提供的，可提供数据（曲率分数）的议案8的特性直接相关的使用和数学为基础的措施简单; （在Restif 等人 8的特征和优点的详细描述）。此外，计算分析使表型特征是不可能的人眼的得分澄清。这里，体现这种分析方法的分辨率数据呈现和易于实现协议描述后续天书分析记录游泳试验。

<table><tbody><tr><td>&lt;strong&gt;REAGENT&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>N2</td><td>Caenorhabditis Genetics Center (CGC)</td><td></td><td>&lt;em&gt;C. elegans&lt;/em&gt; WT (ancestral).</td></tr><tr><td>OP50 &lt;em&gt;Escherichia coli&lt;/em&gt;</td><td>Caenorhabditis Genetics Center (CGC)</td><td></td><td>Biosafety Level: BSL-1.</td></tr><tr><td>OP50-1 &lt;em&gt;Escherichia Coli&lt;/em&gt;&amp;nbsp;</td><td>Caenorhabditis Genetics Center (CGC)</td><td></td><td>Streptomycin resistant strain of OP50.&amp;nbsp; Biosafety Level: BSL-1.</td></tr><tr><td>Streptomycin sulfate salt</td><td>Sigma-Aldrich</td><td>S6501</td><td></td></tr><tr><td>Printed Microscope Slides&amp;nbsp;</td><td>Thermo Fisher Scientific</td><td>Gold Seal Fluorescent Antibody Microslides: 3032-002</td><td>have two etched 10 mm diameter circles delineated by white ceramic ink</td></tr><tr><td>Nematode Growth Medium (NGM)</td><td>For 1 L: 17 g Agar, 3 g NaCl, 2.5 g Peptone, 1 mL 1 M CaCl&lt;sub&gt;2&lt;/sub&gt;, 1 mL 5 mg/mL Cholesterol in ethanol, 25 mL 1 M KPO&lt;sub&gt;4&lt;/sub&gt; buffer, 1 mL 1 M MgSO&lt;sub&gt;4&lt;/sub&gt;, H&lt;sub&gt;2&lt;/sub&gt;O to 1 L. Sterilize by autoclaving.</td><td></td><td>Stiernagle, T. Maintenance of &lt;em&gt;C. elegans&lt;/em&gt;. WormBook, 1-11, doi:10.1895/wormbook.1.101.1 (2006)</td></tr><tr><td>M9 buffer</td><td>For 1 L: 3 g KH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt;, 6 g Na&lt;sub&gt;2&lt;/sub&gt;HPO&lt;sub&gt;4&lt;/sub&gt;, 5 g NaCl, 1 mL 1 M MgSO&lt;sub&gt;4&lt;/sub&gt;, H&lt;sub&gt;2&lt;/sub&gt;O to 1 L. Sterilize by autoclaving.</td><td></td><td>Stiernagle, T. Maintenance of &lt;em&gt;C. elegans&lt;/em&gt;. WormBook, 1-11, doi:10.1895/wormbook.1.101.1 (2006)</td></tr><tr><td>&lt;strong&gt;EQUIPMENT&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>CeleST</td><td>Driscoll Lab, Rutgers University</td><td>&lt;em&gt;C. elegans&lt;/em&gt; Swim Test</td><td>Open Source, see http://celestmod.github.io/CeleST/ and http://celest.mbb.rutgers.edu/</td></tr><tr><td>MATLAB</td><td>www.mathworks.com/downloads</td><td>MatLab version 2015b (best)</td><td>The CeleST version demonstrated here has best functionality with Mac OS 10.10 and MatLab 2015b. MATLAB 2015B introduced changes to how MATLAB handled graphics, including a new coding convention and syntax. These changes resolved an issue that couldn&#039;t be resolved elegantly (primarily because the internals of MATLAB really needed the major graphics overhaul implemented in MATLAB 2015B). For this reason, CeleST should always be run on MATLAB 2015B or later versions. However for users without access to MATLAB version 2015B or later (or MATLAB at all), we have created a CeleST program that doesn&#039;t need MATLAB on the computer at al. An installer is downloaded by the prospective user and then it installs itself onto the computer through a couple prompts like most programs.&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Mac OS</td><td>www.apple.com</td><td>Version 10.10</td><td>Currently, CeleST has been ported to the major operating systems (Windows, Mac, and Linux). The current code can be run on any of the operating systems and there are versions for each operating system that don&#039;t even require users to have MATLAB to use CeleST (this version requires a large download). The Windows version has been tested the least and is most prone to bugs as such. Linux has been moderately tested. And Mac has been and continues to be tested extensively (primarily because it&#039;s the operating system in our lab).</td></tr><tr><td>Stereomicroscope&amp;nbsp;</td><td>Zeiss</td><td>Stemi 2000-C&amp;nbsp;</td><td></td></tr><tr><td>Transmitted Light Base</td><td>Diagnostic Instruments</td><td>TLB 3.1</td><td></td></tr><tr><td>Digital CCD Camera</td><td>QImaging</td><td>Rolera-XR Mono Fast 1394 (ROL-XR-F-M-12)</td><td></td></tr><tr><td>Digital Video Recording Software</td><td>Norpix</td><td>Streampix Version 3.17.2</td><td></td></tr></tbody></table>

automated analysis of c. elegans swim behavior using celest software

解剖调节行为的神经元和神经肌肉电路仍然是生物学的一个重大挑战。该线虫已被证明是在帮助解决这一难题，从激励的技术方法，构建人脑连接组，实际上基本功能模式的具体分子司机脱落光了宝贵的模式生物。已在固体基质进行线虫的行为学研究的大部分。在液体中，动物表现出的行为模式，包括在一个范围在三维速度运动，以及局部的身体动作，如无前形状变化的后卷曲，这引入定量的新挑战。一个简单的程序的步骤，以及使用使线虫的高分辨率分析游泳行为的软件，这里提出。该软件名叫张玉华，将使用跟踪一个专门的电脑程序同时对多个动物，并提供C的新措施线虫液体（游泳）运动。这些措施大多是植根于动物的姿势和基于计算机视觉和模式识别用于数学，没有阈临界值计算需求。软件工具可以用来既在数百只动物从合并的小批量试验评估总体游泳实力和揭示新的表型，即使在充分表征的基因的突变体。的标本适乐时分析，制备简单，技术含量低，被科学界能够适应面广。因此此处所描述的计算方法的使用应有助于行为和行为电路中的线虫模型的更多的理解。

通过在液体（游泳）分析运动，表型不是在固体培养基上显而易见（爬行）可以被阐明。定量游泳运动我们开发了测量的游泳行为八十人的新参数的特定软件。八个最有用的这些参数进行详细在表1中说明。这些参数被命名为波起始速度，身体波数，不对称，拉伸，卷边，行驶速度，笔触和活动指数。研究已经例示了软件的功率限定数百与WT，行为或长寿突变体背景8老化成人的功能下降，并分析了充分研究的长寿突变体年龄-1（hx546）和daƒ-16（mgDƒ50），其中滋生扰乱了正常的胰岛素信号通路的突变。基因年龄-1编码的磷脂酰肌醇3-激酶（PIK3）催化亚基，并且，当它怀有突变hx546，使寿命延长和胁迫抗性27-29。该基因daƒ-16编码的寿命缩短或删除时30-33损害的应激反应一个叉头框O（FOXO）转录因子。 如波起始速度，行驶速度，笔触和活动指数游泳这样的一些参数逐渐随着年龄的增长，即使在良好的遗传背景（ 图1）有所下降。在当前的知识线，长寿命的AGE-1（hx546）突变表明在先进的和极其古老的年龄比WT更剧烈的身体表现。也如预期，短期daƒ-16（mgDƒ50）突变体显示的折衷性能，尤其是在极其古老的年龄。值得注意的是，这是只有在天书计算机视觉和数学算法包，该SUPERIO的审查AGE-1（hx546）突变体中的R游泳演出是在成年期发病检测。 年龄-1（hx546）的结果在年轻成人生活增强的物理性能表明，该突变影响正常发育和/或年轻成人表型在以前没有认识到（ 图1）的方式的事实。 体波数，不对称，舒展，和冰壶参数呈上升趋势随着年龄的野生型和突变体老化成人（ 图2）。有趣的是，该软件的分辨率级别显示更精细的行为特征，如年龄-1（hx546）在其整个寿命和极端旧daƒ-16（mgDƒ50）突变体的无力突变体的持续对称伸展和蜷到相同的程度-age WT和年龄-1（hx546）大人。 除了物理PERFO的整体必然损失rmance由于年龄，每个个体成年通过老化过程显示了独特的级数模式，即使当遗传学和环境几乎同质7。 （通过控制遗传学和环境，这些因素的可能的混杂影响减至最小，拉开随机性的年龄相关变性显著贡献。）相似的遗传背景的同步线虫种群保持在受控环境中仍含有的混合根据自己的特点老龄化不同类别的个体。虽然所有的启动健康成人，有的迅速失去其体质（坏腾空类，C类），而其他人保持旺盛的时间较长时期内（婉约腾空，A级）。因此坏腾空高度似乎比曼妙腾空一个相当短长期保健。 在我们的研究中进一步8详细，优雅的腾空高度保持青春的身体fitnESS通过与更年轻的成年人的游泳曲线比较所观察到的（ 图3,4和5）。这个持续健身相当于在后育龄长寿命年龄-1（hx546）突变体的物理性能（D 11）（ 图1和2）。相反，坏的腾空高度明显失去了他们的身体能力再现之后不久，在执行类似的极端老progericdaƒ-16（mgDƒ50）成人（ 图1 - 4）的水平。这些相似性可以通过比较总可以得出，但是特别签名仔细看看容易引人注目。例如，虽然有两拉伸和卷曲在极端旧野生型和老化的突变体的程度之间的一些相关性（ 图2），这个关系不坏不停观察（ 图4和5），其表现出较高的倾向卷曲了，但不测试的样品中的拉伸。我们目前的软件从而通过提供工具，更复杂的研究，不听话之前添加维体质或运动能力的分析。综上所述，张玉华提供了在这里强调8新措施，其中定义了特定的基因，表观遗传学行为指纹和环境背景的形式全面读数，使独特而通用参数模式的识别，可以根据具体情况的签名（环境，药理，营养），生物过程或生物体状态如长期保健。 <img alt="图1" src="/files/ftp_upload/54359/54359fig1.jpg" /> 图1:波引发速率（A），活动性指数（B），笔触（C）及旅行速度（D）为WT， 年龄-1（hx546）和daƒ-16天书软件报告（mgDƒ50）<html>第D 4（青年期），11（-复制后）和20（极端旧）成人。在y轴上的手段&#39;号&#39;&#39;＃&#39;。的WT在绿色和daƒ-16中的红色着色灰色， 年龄-1。误差条是平均值（SEM）的标准误差。同龄WT和老化的突变体，使用单向方差分析，接着进行Dunnett的多重对比试验统计学意义进行比较。 **，p值= 0.001 - &lt;0.01; ***，P = 0.0001 - &lt;0.001。 62例中的每个数据点从四个独立的试验。注意，这里，和图2中 ，每个单独的30秒的视频与4只动物制成，以及每个试验中，我们得分总共4游泳视频16的动物，这是用于示出每个数据点4生物学重复进行。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54359/54359fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <html> ntent"FO:保together.within页="1"&gt; <img alt="图2" src="/files/ftp_upload/54359/54359fig2.jpg" /> 图2:体波数（A），非对称性（B），拉伸（C），和冰壶（D）为WT， 年龄-1（hx546）和daƒ-16第D 4（mgDƒ50）成人软件报告（年轻成年），11（后再现）和20（极端老）。在y轴上的手段&#39;号&#39;&#39;＃&#39;。 WT在灰色， 年龄-1在绿色和DAF-16中的红色着色。误差条是平均值（SEM）的标准误差。同龄WT和老化的突变体，使用单向方差分析，接着进行Dunnett的多重对比试验统计学意义进行比较。 *，P = 0.01 - &lt;0.05; **，p值= 0.001 - &lt;0.01; ***，P = 0.0001 - &lt;0.001。 N =在每个数据点62从四个独立的，30秒游泳试验。e.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 <img alt="图3" src="/files/ftp_upload/54359/54359fig3.jpg" /> 图3:波引发速率（A），活动性指数（B），笔触（C），及旅行速度（D）为年轻WT大人（D 4），和同龄优美和坏腾空（D10软件报告和11）。在y轴上的手段&#39;号&#39;&#39;＃&#39;。年轻的工作小组是彩色灰色，A级优美的腾空高度的绿色和C类不良腾空高度为红色。误差条是平均值（SEM）的标准误差。 A级优美的腾空高度和C类不良腾空采用单因素方差分析，接着进行Dunnett的多重对比试验相比，到D 4青壮年。 ****，P &lt;0.0001。 N =在来自两个独立的每个数据点27，30秒游泳试验。图稍微偏离Restif 等进行修改。 （2014年）8，这是创意下公布共享署名（CC BY）牌照http://creativecommons.org/licenses/by/4.0/。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54359/54359fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/54359/54359fig4.jpg" /> 图4:体波数（A），非对称性（B），和冰壶（C）软件报告为年轻成人WT（D 4），和同龄优雅和坏腾空（D 10和11）。在y轴上的手段&#39;号&#39;&#39;＃&#39;。年轻的工作小组以灰色，有色类绿色和C类不良腾空从容腾空高度为红色。误差条是平均值（SEM）的标准误差。 A级优美的腾空高度和C类不良腾空采用单因素方差分析，接着进行Dunnett的多重对比试验相比，到D 4青壮年。 **，p值= 0.001 - &lt;0.01; ****，P &lt;0.0001; <s仲&gt; N / A，不适用，因为只有一个动物出来的总样本量的卷曲。 N =在来自两个独立的每个数据点27，30秒游泳试验。图稍微偏离Restif 等进行修改。 （2014年）8，这是在知识共享署名（CC BY）牌照HTTP下出版:/creativecommons.org/licenses/by/4.0/。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54359/54359fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/54359/54359fig5.jpg" /> 图5:拉伸软件报告为年轻的成年人WT（D 4），和同龄优美和坏腾空（D 10和11）。年轻的工作小组以灰色，有色类绿色和C类不良腾空从容腾空高度为红色。误差条是平均值（SEM）的标准误差。 A类优美腾空类和C类不良腾空进行了比较到D 4ÿ翁荣南成人使用单向ANOVA，然后通过Dunnett的多重对比试验。 N =在每个数据27指向来自两个独立试验。图稍微偏离Restif 等进行修改。 （2014年）8，这是在知识共享署名（CC BY）牌照HTTP下出版:/creativecommons.org/licenses/by/4.0/。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54359/54359fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="视频1" src="/files/ftp_upload/54359/54359video1.jpg" /> 视频1:C的一组有代表性的游泳线虫成虫。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54359/Video1.mov" target="_blank">请点击此处观看该视频。 （右键点击下载）。</a> <img alt="视频2" src="/files/ftp_upload/54359/54359video2.jpg" /> 视频2:测试动物的游泳表演的各个曲率地图天书软件计算。曲率映射计算的背景;它们不会出现在与用户的软件接口。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54359/Video2.mov" target="_blank">请点击此处观看该视频。 （右键点击下载）。</a> <img alt="视频3" src="/files/ftp_upload/54359/54359video3.jpg" /> 视频3:根据个人的曲率地图游泳办法计算软件。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54359/Video3.mov" target="_blank">请点击此处观看该视频。 （右键点击下载）。</a> <img alt="视频4" src="/files/ftp_upload/54359/54359video4.jpg" /> 视频4:不曲率依靠游泳办法计算软件地图。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54359/Video4.mov" target="_blank">请点击此处观看该视频。 （右键点击下载）。</a>

Watch this Scientific Journal Video about Automated Analysis of C. elegans Swim Behavior Using CeleST Software at JoVE.com

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

对于在液体线虫游泳行为的基于计算机的分析一个有效和简单的方法进行说明。该方法需要很少或几乎没有投资线虫实验室。所使用的硬件是标准的，并且该计算机软件的行为分析（CELEST）是一个开放源码之一。

的自动分析<em&gt;℃。线虫</em&gt;游泳行为使用天书软件

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has ...

automated-analysis-of-c-elegans-swim-behavior-using-celest-software

Universidad San Sebastian

Research

JoVE Journal

Neuroscience

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

12.4K 次观看.  Rutgers University. 对于在液体线虫游泳行为的基于计算机的分析一个有效和简单的方法进行说明。该方法需要很少或几乎没有投资线虫实验室。所使用的硬件是标准的，并且该计算机软件的行为分析（CELEST）是一个开放源码之一。 

视频: Automated Analysis of C. elegans Swim Behavior Using CeleST Software

Single Wavelength Shadow Imaging of Caenorhabditis elegans Locomotion Including Force Estimates

This study demonstrates an inexpensive and straightforward technique that allows the measurement of physical properties such as position, velocity, acceleration and forces involved in the locomotory behavior of nematodes suspended in a column of water in response to single wavelengths of light. We demonstrate how to evaluate the locomotion of a microscopic organism using Single Wavelength Shadow Imaging (SWSI) using two different examples.
The first example is a systematic and statistically viable study of the average descent of C. elegans in a column of water. For this study, we used living and dead wildtype C. elegans. When we compared the velocity and direction of nematode active movement with the passive descent of dead worms within the gravitational field, this study showed no difference in descent-times. The average descent was 1.5 mm/sec &#177; 0.1 mm/sec for both the live and dead worms using 633 nm coherent light.
The second example is a case study of select individual C. elegans changing direction during the descent in a vertical water column. Acceleration and force are analyzed in this example. This case study demonstrates the scope of other physical properties that can be evaluated using SWSI while evaluating the behavior using single wavelengths in an environment that is not accessible with traditional microscopes. Using this analysis we estimated an individual nematode is capable of thrusting with a force in excess of 28 nN.
Our findings indicate that living nematodes exert 28 nN when turning, or moving against the gravitational field. The findings further suggest that nematodes passively descend in a column of water, but can actively resist the force of gravity primarily by turning direction.

Single Wavelength Shadow Imaging of Caenorhabditis elegans Locomotion Including Force Estimates

The technique presented here measures the path of freely swimming microscopic species using single wavelength exposure. C. elegans are used to demonstrate shadow imaging as an inexpensive alternative to costly microscopes. This technique can be adapted to accommodate various orientations, environments and species to measure direction, speed, acceleration and forces.

单波长阴影成像<em&gt;秀丽隐杆线虫</em&gt;步态包括武力在内的估算

This study demonstrates an inexpensive and straightforward technique that allows the measurement of physical properties such as position, velocity, ...

科研

工程学

Quantitative Locomotion Study of Freely Swimming Micro-organisms Using Laser Diffraction

Soil and aquatic microscopic organisms live and behave in a complex three-dimensional environment. Most studies of microscopic organism behavior, in contrast, have been conducted using microscope-based approaches, which limit the movement and behavior to a narrow, nearly two-dimensional focal field.1 We present a novel analytical approach that provides real-time analysis of freely swimming C. elegans in a cuvette without dependence on microscope-based equipment. This approach consists of tracking the temporal periodicity of diffraction patterns generated by directing laser light through the cuvette. We measure oscillation frequencies for freely swimming nematodes.
Analysis of the far-field diffraction patterns reveals clues about the waveforms of the nematodes. Diffraction is the process of light bending around an object. In this case light is diffracted by the organisms. The light waves interfere and can form a diffraction pattern. A far-field, or Fraunhofer, diffraction pattern is formed if the screen-to-object distance is much larger than the diffracting object. In this case, the diffraction pattern can be calculated (modeled) using a Fourier transform.2
C. elegans are free-living soil-dwelling nematodes that navigate in three dimensions. They move both on a solid matrix like soil or agar in a sinusoidal locomotory pattern called crawling and in liquid in a different pattern called swimming.3 The roles played by sensory information provided by mechanosensory, chemosensory, and thermosensory cells that govern plastic changes in locomotory patterns and switches in patterns are only beginning to be elucidated.4 We describe an optical approach to measuring nematode locomotion in three dimensions that does not require a microscope and will enable us to begin to explore the complexities of nematode locomotion under different conditions.

Microscopic organisms like the free-swimming nematode C. elegans, live and behave in a complex three-dimensional environment. We report on a novel approach that provides analysis of C. elegans using diffraction patterns. This approach consists of tracking the temporal periodicity of diffraction patterns generated by directing laser light through a cuvette.

使用激光衍射自由游泳微生物的定量步态研究

Soil and aquatic microscopic organisms live and behave in a complex three-dimensional environment. Most studies of microscopic organism behavior, in ...

生物工程

Microfluidic-based Electrotaxis for On-demand Quantitative Analysis of Caenorhabditis elegans' Locomotion

The nematode Caenorhabditis elegans is a versatile model organism for biomedical research because of its conservation of disease-related genes and pathways as well as its ease of cultivation. Several C. elegans disease models have been reported, including neurodegenerative disorders such as Parkinson's disease (PD), which involves the degeneration of dopaminergic (DA) neurons 1. Both transgenes and neurotoxic chemicals have been used to induce DA neurodegeneration and consequent movement defects in worms, allowing for investigations into the basis of neurodegeneration and screens for neuroprotective genes and compounds 2,3.
Screens in lower eukaryotes like C. elegans provide an efficient and economical means to identify compounds and genes affecting neuronal signaling. Conventional screens are typically performed manually and scored by visual inspection; consequently, they are time-consuming and prone to human errors. Additionally, most focus on cellular level analysis while ignoring locomotion, which is an especially important parameter for movement disorders.
We have developed a novel microfluidic screening system (Figure 1) that controls and quantifies C. elegans' locomotion using electric field stimuli inside microchannels. We have shown that a Direct Current (DC) field can robustly induce on-demand locomotion towards the cathode ("electrotaxis") 4. Reversing the field's polarity causes the worm to quickly reverse its direction as well. We have also shown that defects in dopaminergic and other sensory neurons alter the swimming response 5. Therefore, abnormalities in neuronal signaling can be determined using locomotion as a read-out. The movement response can be accurately quantified using a range of parameters such as swimming speed, body bending frequency and reversal time.
Our work has revealed that the electrotactic response varies with age. Specifically, young adults respond to a lower range of electric fields and move faster compared to larvae 4. These findings led us to design a new microfluidic device to passively sort worms by age and phenotype 6.
We have also tested the response of worms to pulsed DC and Alternating Current (AC) electric fields. Pulsed DC fields of various duty cycles effectively generated electrotaxis in both C. elegans and its cousin C. briggsae 7. In another experiment, symmetrical AC fields with frequencies ranging from 1 Hz to 3 KHz immobilized worms inside the channel 8.
Implementation of the electric field in a microfluidic environment enables rapid and automated execution of the electrotaxis assay. This approach promises to facilitate high-throughput genetic and chemical screens for factors affecting neuronal function and viability.

Microfluidic-based Electrotaxis for On-demand Quantitative Analysis of Caenorhabditis elegans' Locomotion

A semi-automated micro-electro-fluidic method to induce on-demand locomotion in Caenorhabditis elegans is described. This method is based on the neurophysiologic phenomenon of worms responding to mild electric fields (&#x201C;electrotaxis&#x201D;) inside microfluidic channels. Microfluidic electrotaxis serves as a rapid, sensitive, low-cost, and scalable technique to screen for factors affecting neuronal health.

基于微流体Electrotaxis的，为按需定量分析<em&gt;线虫</em&gt;&#39;LOCOMOTION的

The nematode Caenorhabditis elegans is a versatile model organism for biomedical research because of its conservation of disease-related genes and ...

C. elegans Tracking and Behavioral Measurement

We have developed instrumentation, image processing, and data analysis techniques to quantify the locomotory behavior of C. elegans as it crawls on the surface of an agar plate. For the study of the genetic, biochemical, and neuronal basis of behavior, C. elegans is an ideal organism because it is genetically tractable, amenable to microscopy, and shows a number of complex behaviors, including taxis, learning, and social interaction1,2. Behavioral analysis based on tracking the movements of worms as they crawl on agar plates have been particularly useful in the study of sensory behavior3, locomotion4, and general mutational phenotyping5. Our system works by moving the camera and illumination system as the worms crawls on a stationary agar plate, which ensures no mechanical stimulus is transmitted to the worm. Our tracking system is easy to use and includes a semi-automatic calibration feature. A challenge of all video tracking systems is that it generates an enormous amount of data that is intrinsically high dimensional. Our image processing and data analysis programs deal with this challenge by reducing the worms shape into a set of independent components, which comprehensively reconstruct the worms behavior as a function of only 3-4 dimensions6,7. As an example of the process we show that the worm enters and exits its reversal state in a phase specific manner.

C. elegans Tracking and Behavioral Measurement

We have developed a video-rate tracking microscope system that can record and quantify C. elegans behavior at high resolution and high speeds. We have also developed computational methods to reduce the dimensionality of the worm images to a fundamental set of measurements that completely describe the shape of the worm.

线虫跟踪和行为测量

We have developed instrumentation, image processing, and data analysis techniques to quantify the locomotory behavior of C. elegans as it crawls on the ...

神经科学

Swimming Induced Paralysis to Assess Dopamine Signaling in Caenorhabditis elegans

The swimming assay described in this protocol is a valid tool to identify proteins regulating the dopaminergic synapses. Similar to mammals, dopamine (DA) controls several functions in C. elegans including learning and motor activity. Conditions that stimulate DA release (e.g., amphetamine (AMPH) treatments) or that prevent DA clearance (e.g., animals lacking the DA transporter (dat-1) which are incapable of reaccumulating DA into the neurons) generate an excess of extracellular DA ultimately resulting in inhibited locomotion. This behavior is particularly evident when animals swim in water. In fact, while wild-type animals continue to swim for an extended period, dat-1 null mutants and wild-type treated with AMPH or inhibitors of the DA transporter sink to the bottom of the well and do not move. This behavior is termed "Swimming Induced Paralysis" (SWIP). Although the SWIP assay is well established, a detailed description of the method is lacking. Here, we describe a step-by-step guide to perform SWIP. To perform the assay, late larval stage-4 animals are placed in a glass spot plate containing control sucrose solution with or without AMPH. Animals are scored for their swimming behavior either manually by visualization under a stereoscope or automatically by recording with a camera mounted on the stereoscope. Videos are then analyzed using a tracking software, which yields a visual representation of thrashing frequency and paralysis in the form of heat maps. Both the manual and automated systems guarantee an easily quantifiable readout of the animals' swimming ability and thus facilitate screening for animals bearing mutations within the dopaminergic system or for auxiliary genes. In addition, SWIP can be used to elucidate the mechanism of action of drugs of abuse such as AMPH.

Swimming Induced Paralysis to Assess Dopamine Signaling in Caenorhabditis elegans

Swimming induced paralysis (SWIP) is a well-established behavioral assay used to study the underlying mechanisms of dopamine signaling in Caenorhabditis elegans (C. elegans). However, a detailed method to perform the assay is lacking. Here, we describe a step-by-step protocol for SWIP.

游泳诱导麻痹对线虫多巴胺信号的评价

The swimming assay described in this protocol is a valid tool to identify proteins regulating the dopaminergic synapses. Similar to mammals, dopamine (DA) ...

行为学

Comparative Analysis of Experimental Methods to Quantify Animal Activity in Caenorhabditis elegans Models of Mitochondrial Disease

Caenorhabditis elegans is widely recognized for its central utility as a translational animal model to efficiently interrogate mechanisms and therapies of diverse human diseases. Worms are particularly well-suited for high-throughput genetic and drug screens to gain deeper insight into therapeutic targets and therapies by exploiting their fast development cycle, large brood size, short lifespan, microscopic transparency, low maintenance costs, robust suite of genomic tools, mutant repositories, and experimental methodologies to interrogate both in vivo and ex vivo physiology. Worm locomotor activity represents a particularly relevant phenotype that is frequently impaired in mitochondrial disease, which is highly heterogeneous in causes and manifestations but collectively shares an impaired capacity to produce cellular energy. While a suite of different methodologies may be used to interrogate worm behavior, these vary greatly in experimental costs, complexity, and utility for genomic or drug high-throughput screens. Here, the relative throughput, advantages, and limitations of 16 different activity analysis methodologies were compared that quantify nematode locomotion, thrashing, pharyngeal pumping, and/or chemotaxis in single worms or worm populations of C. elegans at different stages, ages, and experimental durations. Detailed protocols were demonstrated for two semi-automated methods to quantify nematode locomotor activity that represent novel applications of available software tools, namely, ZebraLab (a medium-throughput approach) and WormScan (a high-throughput approach). Data from applying these methods demonstrated similar degrees of reduced animal activity occurred at the L4 larval stage, and progressed in day 1 adults, in mitochondrial complex I disease (gas-1(fc21)) mutant worms relative to wild-type (N2 Bristol) C. elegans. This data validates the utility for these novel applications of using the ZebraLab or WormScan software tools to quantify worm locomotor activity efficiently and objectively, with variable capacity to support high-throughput drug screening on worm behavior in preclinical animal models of mitochondrial disease.

Comparative Analysis of Experimental Methods to Quantify Animal Activity in Caenorhabditis elegans Models of Mitochondrial Disease

This study presents protocols for two semi-automated locomotor activity analysis approaches in C. elegans complex I disease gas-1(fc21) worms, namely, ZebraLab (a medium-throughput assay) and WormScan (a high-throughput assay) and provide comparative analysis among a wide array of research methods to quantify nematode behavior and integrated neuromuscular function.

线粒体疾病脑 膜炎基因 模型中量化动物活动的实验方法比较分析

Caenorhabditis elegans is widely recognized for its central utility as a translational animal model to efficiently interrogate mechanisms and therapies of ...

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the development of more effective treatments is the relative lack of understanding of the molecular underpinnings of the effects of ethanol on behavior. The nematode, Caenorhabditis elegans (C.&#160;elegans), provides a useful model in which to generate and test hypotheses about the molecular effects of ethanol. Here, we describe an assay that has been developed and used to examine the roles of particular genes and environmental factors in behavioral responses to ethanol, in which locomotion is the behavioral output. Ethanol dose-dependently causes an acute depression of crawling on an agar surface. The effects are dynamic; animals exposed to a high concentration demonstrate an initial strong depression of crawling, referred to here as initial sensitivity, and then partially recover locomotion speed despite the continued presence of the drug. This ethanol-induced behavioral plasticity is referred to here as the development of acute functional tolerance. This assay has been used to demonstrate that these two phenotypes are distinct and genetically separable. The straightforward locomotion assay described here is suitable for examining the effects of both genetic and environmental manipulations on these acute behavioral responses to ethanol in C.&#160;elegans.

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

C. elegans is a useful model for studying the effects of ethanol on behavior. We present a behavioral assay that quantifies the effects of ethanol on the locomotion speed of crawling worms; both initial sensitivity and the development of acute functional tolerance to ethanol can be measured with this assay.

测量乙醇的影响上的步态速度的测定<em&gt;秀丽隐杆线虫</em

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the ...

Automated Behavioral Analysis of Large C. elegans Populations Using a Wide Field-of-view Tracking Platform

Caenorhabditis elegans is a well-established animal model in biomedical research, widely employed in functional genomics and ageing studies. To assess the health and fitness of the animals under study, one typically relies on motility readouts, such as the measurement of the number of body bends or the speed of movement. These measurements usually involve manual counting, making it challenging to obtain good statistical significance, as time and labor constraints often limit the number of animals in each experiment to 25 or less. Since high statistical power is necessary to obtain reproducible results and limit false positive and negative results when weak phenotypic effects are investigated, efforts have recently been made to develop automated protocols focused on increasing the sensitivity of motility detection and multi-parametric behavioral profiling. In order to extend the limit of detection to the level needed to capture the small phenotypic changes that are often crucial in genetic studies and drug discovery, we describe here a technological development that enables the study of up to 5,000 individual animals simultaneously, increasing the statistical power of the measurements by about 1,000-fold compared to manual assays and about 100-fold compared to other available automated methods.

Automated Behavioral Analysis of Large C. elegans Populations Using a Wide Field-of-view Tracking Platform

We describe protocols for using the wide field-of-view nematode tracking platform (WF-NTP), which enables high-throughput phenotypic characterization of large populations of Caenorhabditis elegans. These protocols can be used to characterize subtle behavioral changes in mutant strains or in response to pharmacological treatment in a highly scalable fashion.

利用广域跟踪平台对大型线虫种群进行自动行为分析

Caenorhabditis elegans is a well-established animal model in biomedical research, widely employed in functional genomics and ageing studies. To assess the ...

免疫与感染

Evaluation of Motor Impairment in C. elegans Models of Amyotrophic Lateral Sclerosis

The neurodegenerative disease amyotrophic lateral sclerosis (ALS) features progressive loss of motor neurons accompanied by muscle weakness and motor impairment that worsens with time. While considerable advances have been made in determining genetic drivers of ALS for a subset of patients, the majority of cases have an unknown etiology. Further, the mechanisms underlying motor neuron dysfunction and degeneration are not well understood; therefore, there is an ongoing need to develop and characterize representative models to study these processes. Caenorhabditis elegans can adapt their movement to the physical constraints of their surroundings, with two primary movement paradigms studied in a laboratory environment- crawling on a solid surface and swimming in liquid. These represent a complex interplay between sensation, motor neurons, and muscles. C. elegans models of ALS can exhibit impairment in one or both of these movement paradigms. This protocol describes two sensitive assays for evaluating motility in C. elegans: an optimized radial locomotion assay measuring crawling on a solid surface and an automated method for tracking and analyzing swimming in liquid (thrashing). In addition to the characterization of baseline motor impairment of ALS models, these assays can detect suppression or enhancement of the phenotypes from genetic or small molecule interventions. Thus, these methods have utility for studying ALS&#160;models and any C. elegans strain that exhibits altered motility.

Evaluation of Motor Impairment in C. elegans Models of Amyotrophic Lateral Sclerosis

This protocol describes two sensitive assays for discriminating among mild, moderate, and severe motor impairment in C. elegans models of amyotrophic lateral sclerosis, with general utility for C. elegans strains, with altered motility.

秀丽隐杆线虫运动障碍评估 肌萎缩性侧索硬化症模型

The neurodegenerative disease amyotrophic lateral sclerosis (ALS) features progressive loss of motor neurons accompanied by muscle weakness and motor ...

Swimming-Induced Paralysis (SWIP) Assay: A Method to Quantify Dopamine-Mediated Locomotion in C. elegans

Source: Kudumala, S., et al. <a href="https://www-jove-com.bdigitaluss.remotexs.co/video/59243/swimming-induced-paralysis-to-assess-dopamine-signaling">Swimming Induced Paralysis to Assess Dopamine Signaling in&#160;Caenorhabditis elegans</a>.&#160;J. Vis. Exp.&#160;(2019).
Here, we introduce an assay to measure SWIP, a behavioral phenotype of&#160;C. elegans&#160;that occurs during vigorous motion like swimming.&#160; The example protocol discusses both manual and automated analysis approaches.

游泳诱导麻痹 （SWIP） 测定:一种量化秀丽隐杆线虫多巴胺介导的运动的方法

Source: Kudumala, S., et al. Swimming Induced Paralysis to Assess Dopamine Signaling in&#160;Caenorhabditis elegans.&#160;J. Vis. Exp.&#160;(2019).
Here, ...

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单波长阴影成像

使用激光衍射自由游泳微生物的定量步态研究

基于微流体Electrotaxis的，为按需定量分析

线虫跟踪和行为测量

游泳诱导麻痹对线虫多巴胺信号的评价

线粒体疾病脑膜炎基因模型中量化动物活动的实验方法比较分析

测量乙醇的影响上的步态速度的测定

利用广域跟踪平台对大型线虫种群进行自动行为分析

秀丽隐杆线虫运动障碍评估肌萎缩性侧索硬化症模型

游泳诱导麻痹（SWIP）测定:一种量化秀丽隐杆线虫多巴胺介导的运动的方法

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摘要

探索更多视频

此视频中的章节

单波长阴影成像

使用激光衍射自由游泳微生物的定量步态研究

基于微流体Electrotaxis的，为按需定量分析

线虫跟踪和行为测量

游泳诱导麻痹对线虫多巴胺信号的评价

线粒体疾病脑 膜炎基因 模型中量化动物活动的实验方法比较分析

测量乙醇的影响上的步态速度的测定

利用广域跟踪平台对大型线虫种群进行自动行为分析

秀丽隐杆线虫运动障碍评估 肌萎缩性侧索硬化症模型

游泳诱导麻痹 （SWIP） 测定:一种量化秀丽隐杆线虫多巴胺介导的运动的方法

线粒体疾病脑膜炎基因模型中量化动物活动的实验方法比较分析

秀丽隐杆线虫运动障碍评估肌萎缩性侧索硬化症模型

游泳诱导麻痹（SWIP）测定:一种量化秀丽隐杆线虫多巴胺介导的运动的方法