我们感谢Deok Joong Lee对手稿的批判性阅读和测试Python代码。本研究由韩国国家研究基金会2017R1A5A2015369（K.-h.Y.）和2019R1C1C1008708（K.-h.Y.）赞助。

1. 设置成像设备（图 1A-E）
<ol><li>确保具有以下最低要求的智能手机摄像头可用： 12 百万像素 （MP） 摄像头 1080p 分辨率视频 5 GB 存储空间（20 分钟视频为 3-4 GB） 来自应用商店的延时视频应用程序（提供免费应用程序）</li>
	<li>将LED灯箱放在将进行测定的25°C培养箱的底部架上。</li>
	<li>要隐藏 LED 灯表面上的虚线图案，请铺两张纸巾以覆盖 LED 盒子的整个表面。</li>
	<li>为标本制作一个升高的载物台（图1A，D）。高架舞台是由空心矩形隧道支撑的透明塑料板。隧道的功能类似于聚光镜来聚焦光线，为标本提供更好的对比度（图 1C）。确保隧道的墙壁有些暗，以尽量减少光散射。这项研究使用了牛皮纸盒。隧道的尺寸为5.5厘米x 17厘米x 4.5厘米（宽x长x高）。LED灯箱最多可容纳五个隧道。</li>
	<li>在载物台上方放置另一个机架以放置电话进行录制（图1B，E）。每部手机将记录三到六个板（一到两排三个板），因此请相应地调整机架高度。这将在标本上方约15厘米处（图1B）。</li>
	<li>在培养箱内放置一个电源板，以便在过夜录制期间插入手机。</li>
</ol>2. 缓冲液和培养基的制备
<ol><li>通过将 3 g KH 2 PO 4、6 g Na 2 HPO4 和 5 g NaCl 加入 1 L 蒸馏的 H2O 中来制备 M9 缓冲液。冷却缓冲液，然后加入 1 mL 的 1 M MgSO4。</li>
	<li>通过将 108.3 g KH 2 PO 4 和 35.6 g K 2 HPO 4 添加到 1 L H2O 中制备 1 M KPO4 缓冲液。通过高压灭菌灭菌。</li>
	<li>通过混合 1 mL 漂白剂、0.4 mL 1 M NaOH 和 2.6 mL H2O 来制备蠕虫漂白溶液。</li>
	<li>准备线虫生长培养基（NGM）琼脂平板。<ol><li>在 3 L 烧瓶中加入 3 g NaCl、2.5 g 杆菌蛋白胨和 17 g 杆菌琼脂。加入 975 mL 蒸馏水并插入搅拌棒。</li>
		<li>高压灭菌，然后冷却至55°C，并加入1mL胆固醇（5mg / mL乙醇溶液），1mL1M CaCl2，1mL1M MgSO4和25mL1M KPO4 缓冲液（pH 6.0）。搅拌均匀。倒入6厘米的盘子中。让板干燥至少 2 天。</li>
	</ol></li>
	<li>通过移液约 1 mL 的 OP50 过夜培养物形成细菌草坪，用 OP50 大肠杆菌 接种 NGM 琼脂平板。在室温 （RT） 下放置，直到准备使用。</li>
</ol>3. 高蛋白胨NGM板的制备（用于PA14）
注意：这些板应在测定前至少5天制作。
<ol><li>制作含有0.35%蛋白胨的NGM。在 250 mL 锥形瓶中混合 0.3 g NaCl、0.35 g 杆菌蛋白胨和 1.7 g 杆菌琼脂。加入 97.5 mL 蒸馏水并插入搅拌棒。</li>
	<li>用铝箔盖住烧瓶口，并在121°C高压灭菌20分钟。</li>
	<li>冷却至 55 °C，加入 0.1 mL 胆固醇（5 mg/mL 乙醇溶液）、0.1 mL 1 M CaCl 2、0.1 mL 1 M MgSO 4 和2.5 mL 1 M KPO4 缓冲液 （pH 6.0）。搅拌均匀。</li>
	<li>将高蛋白胨NGM倒入35毫米培养皿中。</li>
	<li>将盘子干燥至少 2 天。</li>
</ol>4. 通过漂白同步蠕虫
注意：在测定前3天开始此步骤。
<ol><li>取装有妊娠成虫的平板，并通过用 M9 缓冲液洗涤平板将它们收集到 1.7 mL 微管中。</li>
	<li>尽可能多地去除液体，然后加入 400 μL 漂白溶液。等待约4-5分钟，间歇性涡旋，直到成虫体破裂，释放卵。</li>
	<li>加入M9缓冲液以填充微管的其余部分以稀释漂白溶液。以最大速度（12，000 至 13，000 x g）旋转 1-2 秒。除去上清液并用M9缓冲液再洗涤三次。</li>
	<li>将鸡蛋转移到含有M9缓冲液的空35mm培养皿中。让卵在20°C下孵化过夜。 在没有食物的情况下，孵化的蠕虫将在L1幼虫阶段停止，使所有蠕虫的发育阶段同步。 注意：用明胶溶液（高压灭菌水中的0.05%明胶）涂覆35毫米培养皿可以防止鸡蛋粘在底部并最大限度地减少鸡蛋损失。</li>
	<li>第二天，将L1阶段蠕虫转移到OP50播种的NGM板上。</li>
	<li>将蠕虫在20°C孵育53-54小时，直到蠕虫达到L4幼虫阶段。</li>
</ol>5. 细菌的制备（铜绿假单胞菌，PA14）
注意：在测定前4天开始此步骤。
<ol><li>在没有任何抗生素的Luria Bertani（LB）琼脂平板上从-80°C划线解冻的细菌，并在37°C下孵育过夜。 注意：始终使用新鲜细菌。条纹板应在4°C下储存不超过1周。</li>
	<li>将单个菌落接种到3mL的King's肉汤中，并在37°C振荡培养箱中生长过夜。</li>
	<li>第二天，将7μL过夜培养物接种到高蛋白胨NGM板上，并在37°C下孵育24小时。</li>
	<li>将接种板移至室温，并在使用前再孵育24小时。准备好后，在接下来的24小时内使用板。</li>
</ol>6. 准备录制
注意：在测定之前执行此操作。
<ol><li>将智能手机插入连接到电源插座的配电盘。确保禁用自动锁定设置，以防止手机在录制时返回锁定屏幕。</li>
	<li>打开延时相机应用程序并将延时摄影间隔设置为 2 秒。将视频质量设置为 1080 fps 的 30p。</li>
	<li>将智能手机屏幕朝上放置，以使用后置摄像头进行录制。检查屏幕以确保纸盒隧道适合视野。</li>
</ol>7. 草坪回避测定
<ol><li>使用铂线镐将30个同步的L4级蠕虫（从L1开始53-54小时）转移到PA14板上。将蠕虫放在细菌草坪的中间。对于本研究中的每个条件，测试了两个板（即每个条件60个蠕虫）。</li>
	<li>将两个板放在录音设备的升高舞台上，盖子朝下。琼脂的一侧将朝上朝向相机。</li>
	<li>在智能手机屏幕上，点击板的位置，以便相机可以聚焦在检测板上。在板上贴上标签或文字会有所帮助，因为相机可以使用它来正确对焦。 注意：在板底部书写不会干扰蠕虫的成像，只要它朝向边缘。幸运的是，蠕虫即使在离开后也会留在草坪附近，因此只需要一览无余的草坪周围区域。</li>
	<li>开始录制。</li>
	<li>录制开始后，向舞台添加更多板。由于通过拾取转移蠕虫所需的时间，板之间可能存在明显的滞后时间。请注意之后的滞后时间，以便每个条件都可以在开始时计算。</li>
	<li>从放置在舞台上的最后一组板记录20小时。在最后的延时视频中，20小时的录制将产生20分钟长的视频。 注意：在测定后直接从平板上计数蠕虫可能是值得的，至少在最初几次开始时。这可以与通过视频成像获得的值进行比较，以确保它们产生相似的数字。</li>
</ol>8. 使用Python脚本处理视频
<ol><li>将电影文件传输到计算机进行处理。扩展名将是MOV（iPhone）或MP4文件（Android）。</li>
	<li>使用 Python 代码处理视频。代码可以在 github.com/khyoon201/wormavoid 找到。</li>
	<li>要运行 Python 脚本，请确保计算机上预安装了以下内容：ffmpeg，一种用于转换视频文件的工具（安装说明可以在其网站上找到，ffmpeg.org/download），以及 Python 包 os、pandas、tkinter 和 ffmpeg-python。</li>
	<li>使用 extract_frame.py 脚本查找每个板的尺寸和坐标。<ol><li>运行 extract_frame.py 脚本。将出现一个窗口以选择存储在计算机上的视频文件。运行完成后，同一目录中会出现一个同名的 jpeg 文件。</li>
		<li>在 ImageJ （imagej.org） 中打开 jpeg 文件。</li>
		<li>从菜单中选择 “分析>设置测量值”。确保选中 “显示标签 ”框（图2A）。关闭窗口。</li>
		<li>使用“ 直线” 工具，通过在板上画一条线来测量板的直径，然后从菜单中选择 “分析>测量 ”。如果视频为 1080p，则每个板的宽度约为 480 像素。记下此信息并关闭 “结果 ”窗口。</li>
		<li>使用多点工具，标记每个板左上角的 点 。这些点将成为裁剪视频的左上角（图 2B）。顺序很重要;按板启动时间的顺序标记。为所有板创建一个点后，从菜单中选择 分析>测量 。测量值（包括点的 X 和 Y 坐标）将显示在“结果”窗口中。</li>
		<li>要处理多个视频，请在 ImageJ 中对其他 jpeg 文件重复此过程。所有 X 和 Y 坐标将列在同一 “结果 ”窗口中。</li>
		<li>将 “结果 ”窗口保存到 csv 文件中。该文件应保存到与影片文件相同的目录中。</li>
	</ol></li>
	<li>找到每个盘子的开始时间。<ol><li>在电脑或手机上播放电影，并记下放置在相机下方的每组板的开始时间。</li>
		<li>打开包含坐标的 Results.csv 文件并添加“开始”列。对于与单个板对应的每一行，在“开始”列下输入适当的开始时间（以秒为单位）（例如，如果开始时间为 0：00：08，请输入 8）。救。 注意：列名称必须为“开始”（小写，不带引号），以便下一个脚本识别以进行裁剪和修剪。</li>
	</ol></li>
	<li>裁剪和修剪视频。<ol><li>运行 crop_n_trim.py 脚本。</li>
		<li>出现提示时，选择 “结果.csv ”文件。 注意：确保 结果.csv 文件和所有影片文件位于同一目录中。</li>
		<li>输入板尺寸。输入前面记下的像素值。 注意：脚本现在将读取 Results.csv 文件的每一行，通过读取“标签”列中的文件名来查找正确的电影文件，并根据“X”和“Y”列中指示的坐标进行裁剪。每个板块的开始时间将由“开始”列中指示的时间确定。脚本完成运行后，将出现一个与电影同名的文件夹，后跟开始时间（例如“Movie1_8”），其中将保存与测定的每个小时时间点对应的 10 秒视频。</li>
	</ol></li>
</ol>9. 使用 ImageJ 手动计数
<ol><li>在 ImageJ 中打开每个 AVI 文件。</li>
	<li>数一数草坪外可见的蠕虫。在一个帧中重叠的蠕虫通常会在另一个帧中分开，以便可以正确计数。</li>
	<li>计算每个时间点的入住率： 占用率=（蠕虫总数-草坪外蠕虫数量）/蠕虫总数 注意：在视频播放过程中，蠕虫会进出草坪，但这不会显著改变结果。尝试使用似乎是平均值的数字，或者确切的每小时时间点（视频中的 5 秒）的蠕虫数量。</li>
</ol>

用于 秀丽隐杆线 虫草坪回避测定的基于智能手机的成像

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未声明利益冲突。

成像动物行为，而不是依靠直接观察，不仅方便，而且具有留下视觉文档的优点。这允许客观的第三人进行盲分析，甚至可以使用图像识别技术进行自动分析。尽管有这些优点，但通常提供的标准设备成本很高，因此一旦购买，就会致力于设置。
使用智能手机收集简单 秀丽隐杆线虫 行为的视频记录有几个优点。它需要对技术知识的最小熟悉程度，并且非常容易设置，...

在研究秀丽隐杆线虫的众多优点中，其简单的神经系统为研究遗传和细胞水平的变化如何影响网络功能和行为输出提供了机会。尽管神经元数量有限，秀丽隐杆线虫表现出广泛的复杂行为。其中之一是避免草坪，其中细菌线虫通过离开细菌草坪来响应有害食物来源。秀丽隐杆线虫避开致病细菌1，2，3 的草坪、产生毒素或掺有毒素的细菌草坪1，4，甚至是表达 RNAi 的细菌，其靶基因敲低对蠕虫的健康有害4，5。研究表明，蠕虫对外部线索做出反应，例如致病菌产生的代谢物1，6，或表明食物使它们生病的内部线索4，7。这些线索通过保守的信号通路处理，例如丝裂原活化蛋白激酶（MAPK）通路和转化生长因子β（TGFβ）通路，并且需要肠道和神经系统之间的通信4，6，7，8。
虽然测定很简单，但学习行为会在几个小时内发展，通常是在一夜之间。虽然有些突变体无法离开，在这种情况下，仅在一个时间点回避得分就足以证明缺陷，但许多突变体最终会离开，但出来的速度较慢。对于这些，需要每隔几个小时跟踪一次蠕虫的运动，这可能很难在一夜之间完成。计数本身也需要时间，在板之间产生滞后时间，从而限制了可以同时测试的板的数量。在整个测定过程中使用成像设置同时记录许多板将非常有用，但设置成本可能令人望而却步，具体取决于研究实验室的资金情况。
为了解决这个问题，我们设计了一种非常简单的方法，使用智能手机记录回避测定。每部手机可以录制多达六个检测板的延时视频。为了提供透射光，我们使用可以在线轻松购买的发光二极管（LED）灯箱。检测板放置在高架平台上，由空心矩形隧道支撑，聚焦入射光，产生对比。我们还提供了一个 Python 脚本，可将视频转换为音频视频交错 （AVI） 文件，显示每小时 10 秒的剪辑。然后将视频裁剪到单独的板上，并保存在单独的文件中，以用于手动计数。
该方法提供了一种低成本的程序，也非常易于使用，使用大多数人都容易获得的物品。在这里，我们描述了使用成熟的草坪避免测定法针对人类病原体 铜绿假单胞 菌（PA14）的方法，其方案先前已描述过2，9。最后，我们还回顾了成像方法的注意事项和局限性，适用于那些希望将其应用于其他 秀丽隐杆线虫 行为实验的人。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>35 mm Petri dish</td>
			<td>SPL</td>
			<td>#10035</td>
			<td></td>
		</tr>
		<tr>
			<td>Bacto agar</td>
			<td>BD</td>
			<td>#214010</td>
			<td></td>
		</tr>
		<tr>
			<td>Bacto Peptone</td>
			<td>BD</td>
			<td>#211677</td>
			<td></td>
		</tr>
		<tr>
			<td>CaCl2</td>
			<td>DAEJUNG</td>
			<td>2507-1400</td>
			<td></td>
		</tr>
		<tr>
			<td>Cholesterol</td>
			<td>BioBasic</td>
			<td>CD0122</td>
			<td></td>
		</tr>
		<tr>
			<td>Dipotassium hydrogen phosphate (K2HPO4)</td>
			<td>JUNSEI</td>
			<td>84120-0350</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycerol</td>
			<td>BioBasic</td>
			<td>GB0232</td>
			<td></td>
		</tr>
		<tr>
			<td>King B Broth</td>
			<td>MB cell</td>
			<td>MB-K0827</td>
			<td></td>
		</tr>
		<tr>
			<td>LED light box multi-pad</td>
			<td>Artmate</td>
			<td>N/A</td>
			<td>This is a USB powered, LED light pad for tracing and drawing purposes. Artmate is a Korean brand, but searching for &#34;LED light box for tracing&#34; in any search engine should yield numerous options from other brands. Overall dimension is around 9&#34; x 12&#34; (A4 size). For example, from amazon US: https://www.amazon.com/LITENERGY-Ultra-Thin-Adjustable-Streaming-Stenciling/dp/B07H7FLJX1/ref=sr_1_5?crid=YMYU0VYY226R&#38;keywords= 
			LED%2Blight%2Bbox&#38;qid=1674183224&#38;sprefix 
			=led%2Blight%2Bbo%2Caps%2C270&#38;sr=8-5&#38;th=1</td>
		</tr>
		<tr>
			<td>MgSO4</td>
			<td>DAEJUNG</td>
			<td>5514-4400</td>
			<td></td>
		</tr>
		<tr>
			<td>Plastic paper sleeve (clear)</td>
			<td>Smead</td>
			<td>#85753</td>
			<td>Any clear plastic sheet with a bit of stiffness can be used as stage. For example, from Amazon US: https://www.amazon.com/Smead-Organized-Translucent-Project-85753/dp/B07HJTRCT7/ref=psdc_1069554_t3_B09J48GXQ 
			8</td>
		</tr>
		<tr>
			<td>Potassium dihydrogen phosphate (KH2PO4)</td>
			<td>JUNSEI</td>
			<td>84185-0350</td>
			<td></td>
		</tr>
		<tr>
			<td>Power strip&#160;</td>
			<td></td>
			<td></td>
			<td>To accommodate 3 phones and one LED box, you need at least 4 outlets.</td>
		</tr>
		<tr>
			<td>Smartphone</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Minimum requirement: 12MP wide camera, 1080p HD video recording at 30fps</td>
		</tr>
		<tr>
			<td>Sodium chloride(NaCl)</td>
			<td>DAEJUNG</td>
			<td>#7548-4100</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium phosphate dibasic anhydrous (Na2HPO4)</td>
			<td>YAKURI</td>
			<td>#31727</td>
			<td></td>
		</tr>
	</tbody>
</table>

a smartphone-based imaging method for c. elegans lawn avoidance assay

当暴露于有毒或致病细菌时，线虫秀 丽隐杆线 虫表现出习得的草坪回避行为，其中蠕虫逐渐离开其食物来源并宁愿留在细菌草坪之外。该测定是测试蠕虫感知外部或内部线索以正确应对有害条件的能力的简单方法。虽然检测方法简单，但计数非常耗时，尤其是对于多个样品，并且跨夜的检测持续时间对研究人员来说很不方便。可以长时间对许多板进行成像的成像系统很有用，但成本很高。在这里，我们描述了一种基于智能手机的成像方法来记录 秀丽隐杆线虫的草坪回避。该方法只需要一部智能手机和一个发光二极管（LED）灯箱作为透射光源。使用免费的延时相机应用程序，每部手机最多可以成像六个板，具有足够的清晰度和对比度来手动计数草坪外的蠕虫。生成的电影在每个小时时间点被处理成 10 秒的音频视频交错 （AVI） 文件，然后裁剪以显示每个单板，使其更适合计数。对于那些希望检查回避缺陷的人来说，这种方法是一种具有成本效益的方法，并且有可能扩展到其他 秀丽隐杆线虫 测定。

脚本产生的第一个视频是从测定开始1小时。0小时的视频不会被保存，因为蠕虫在草坪内开始测定，因此占用率始终为100%。
将野生型N2蠕虫与npr-1突变体进行比较，其草坪回避缺陷在文献中已得到充分证实6，10（图3A-E）。在野生类型中可以看出，蠕虫逐渐离开细菌草坪并留在室...

Watch this Scientific Journal Video about A Smartphone-Based Imaging Method for C. elegans Lawn Avoidance Assay at JoVE.com

作者聚焦：一种基于智能手机的秀丽隐杆线虫草坪回避测定成像方法  

本文介绍了一种简单、低成本的方法来记录 秀丽隐杆线虫的草坪回避行为，使用智能手机和发光二极管 （LED） 灯箱等现成的物品。我们还提供了一个 Python 脚本，将视频文件处理成更适合计数的格式。

一种基于智能手机的秀 丽隐杆线虫 草坪回避测定成像方法

When exposed to toxic or pathogenic bacteria, the nematode Caenorhabditis elegans displays a learned lawn avoidance behavior, in which the worms gradually ...

在秀丽隐杆线虫表现出的众多行为中，避免致病草坪是蠕虫的防御策略，以尽量减少其与有害病原体的接触。在我们的实验室中，我们使用这种行为作为工具来识别介导这种行为的神经元正常运作所涉及的基因。草坪回避行为是一种简单的测定，但它会在数小时内发展。因此，如果可以记录测定以观察长时间的行为，则很方便。此外，如果可以同时记录多个分析，那就更好了。尽管文献中描述了许多成像解决方案，但即使是一些价格适中的相机，对于资源有限的实验室来说也可能遥不可及。此外，有些人可能想尝试这种测定，但不打算长期进行。对于这种情况，诸如此协议之类的短期解决方案可能会有所帮助。我们惊人的协议提供了一种简单、低成本的方式来记录蠕虫行为。我们使用智能手机记录测定，然后通过Python脚本处理视频，使其适合计数。

a-smartphone-based-imaging-method-for-c-elegans-lawn-avoidance-assay

Research

JoVE Journal

Behavior

A Smartphone-Based Imaging Method for C. elegans Lawn Avoidance Assay

9.6K 次观看.  Yonsei University Wonju College of Medicine. 在秀丽隐杆线虫表现出的众多行为中，避免致病草坪是蠕虫的防御策略，以尽量减少其与有害病原体的接触。在我们的实验室中，我们使用这种行为作为工具来识别介导这种行为的神经元正常运作所涉及的基因。草坪回避行为是一种简单的测定，但它会在数小时内发展。因此，如果可以记录测定以观察长时间的行为，则很方便。此外，如果可以同时记录多个分析，那就?...

视频: 作者聚焦：一种基于智能手机的秀丽隐杆线虫草坪回避测定成像方法  

Evaluation of a Smartphone-based Human Activity Recognition System in a Daily Living Environment

An evaluation method that includes continuous activities in a daily-living environment was developed for Wearable Mobility Monitoring Systems (WMMS) that attempt to recognize user activities. Participants performed a pre-determined set of daily living actions within a continuous test circuit that included mobility activities (walking, standing, sitting, lying, ascending/descending stairs), daily living tasks (combing hair, brushing teeth, preparing food, eating, washing dishes), and subtle environment changes (opening doors, using an elevator, walking on inclines, traversing staircase landings, walking outdoors). 
To evaluate WMMS performance on this circuit, fifteen able-bodied participants completed the tasks while wearing a smartphone at their right front pelvis. The WMMS application used smartphone accelerometer and gyroscope signals to classify activity states. A gold standard comparison data set was created by video-recording each trial and manually logging activity onset times. Gold standard and WMMS data were analyzed offline. Three classification sets were calculated for each circuit: (i) mobility or immobility, ii) sit, stand, lie, or walking, and (iii) sit, stand, lie, walking, climbing stairs, or small standing movement. Sensitivities, specificities, and F-Scores for activity categorization and changes-of-state were calculated.
The mobile versus immobile classification set had a sensitivity of 86.30% &#177; 7.2% and specificity of 98.96% &#177; 0.6%, while the second prediction set had a sensitivity of 88.35% &#177; 7.80% and specificity of 98.51% &#177; 0.62%. For the third classification set, sensitivity was 84.92% &#177; 6.38% and specificity was 98.17 &#177; 0.62. F1 scores for the first, second and third classification sets were 86.17 &#177; 6.3, 80.19 &#177; 6.36, and 78.42 &#177; 5.96, respectively. This demonstrates that WMMS performance depends on the evaluation protocol in addition to the algorithms. The demonstrated protocol can be used and tailored for evaluating human activity recognition systems in rehabilitation medicine where mobility monitoring may be beneficial in clinical decision-making.

A standardized evaluation method was developed for Wearable Mobility Monitoring Systems (WMMS) that includes continuous activities in a realistic daily living environment. Testing with a series of daily living activities can decrease activity recognition sensitivity; therefore, realistic testing circuits are encouraged for valid evaluation of WMMS performance.

在日常生活环境评价一个智能手机为基础的人类活动识别系统

An evaluation method that includes continuous activities in a daily-living environment was developed for Wearable Mobility Monitoring Systems (WMMS) that ...

科研

行为学

Novel Object Exploration as a Potential Assay for Higher Order Repetitive Behaviors in Mice

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals. RRBs are commonly split into lower-order and higher-order components, with lower order RRBs consisting of motor stereotypies and higher order RRBs consisting of perseverative and sequencing behaviors. Higher order RRBs are challenging to model in mice. Current assays for RRBs in mice focus primarily on the lower order components, making basic biomedical research into potential treatments or interventions for higher-order RRBs difficult. Here we describe a new assay, novel object exploration. This assay uses a basic open-field arena with four novel objects placed around the perimeter. The test mouse is allowed to freely explore the arena and the order in which the mouse investigates the novel objects is recorded. From these data, patterned sequences of exploration can be identified, as can the most preferred object for each mouse. The representative data shared here and past results using the novel object exploration assay illustrate that inbred mouse strains do demonstrate different behavior in this assay and that strains with elevated lower order RRBs also show elevated patterned behavior. As such, the novel object exploration assay appears to possess good face validity for higher order RRBs in humans and may be a valuable assay for future studies investigating novel therapeutics for ASD.

Higher order restricted, repetitive behaviors (RRBs) disrupt the lives of affected individuals. These behaviors are challenging to model in rodents, making basic biomedical research into potential treatments or interventions for RRBs difficult. Here we describe novel object exploration as a potential assay for higher order RRBs in mice.

新对象的探索在小鼠电位测定高阶重复的行为

Restricted, repetitive behaviors (RRBs) are a core feature of autism spectrum disorder (ASD) and disrupt the lives of affected individuals.  RRBs are ...

High-Throughput Method for Measuring Alcohol Sedation Time of Individual Drosophila melanogaster

Drosophila melanogaster provides an excellent model to study the genetic underpinnings of alcohol sensitivity. In contrast to studies in human populations, the Drosophila model allows strict control over genetic background, and virtually unlimited numbers of individuals of the same genotype can be reared rapidly under well-controlled environmental conditions without regulatory restrictions and at relatively low cost. Flies exposed to ethanol undergo physiological and behavioral changes that resemble human alcohol intoxication, including loss of postural control, sedation, and development of tolerance. Here, we describe a simple, low-cost, high-throughput assay for assessing alcohol sedation sensitivity in large numbers of single flies. The assay is based on video recording of single flies introduced without anesthesia in 24-well cell culture plates in a set-up that enables synchronous initiation of alcohol exposure. The system enables a single person to collect individual ethanol sedation data on as many as 2,000 flies within an 8 h work period. The assay can, in principle, be extended to assess the effects of exposure to any volatile substance and applied to measure effects of acute toxicity of volatiles on other insects, including other fly species.

High-Throughput Method for Measuring Alcohol Sedation Time of Individual Drosophila melanogaster

Current methods to measure alcohol sensitivity in Drosophila are designed to test groups of flies. We present a simple, low-cost, high-throughput assay for assessing alcohol sedation sensitivity in large numbers of single flies. The method does not require specialized tools and can be performed in any laboratory using common materials.

测量个体果蝇黑色素气器酒精镇分时间的高通量方法

Drosophila melanogaster provides an excellent model to study the genetic underpinnings of alcohol sensitivity. In contrast to studies in human ...

Calcium Imaging in Freely Behaving Caenorhabditis elegans with Well-Controlled, Nonlocalized Vibration

Nonlocalized mechanical forces, such as vibrations and acoustic waves, influence a wide variety of biological processes from development to homeostasis. Animals cope with these stimuli by modifying their behavior. Understanding the mechanisms underlying such behavioral modification requires quantification of neural activity during the behavior of interest. Here, we report a method for calcium imaging in freely behaving Caenorhabditis elegans with nonlocalized vibration of specific frequency, displacement, and duration. This method allows the production of well-controlled, nonlocalized vibration using an acoustic transducer and quantification of evoked calcium responses at single-cell resolution. As a proof of principle, the calcium response of a single interneuron, AVA, during the escape response of C. elegans to vibration is demonstrated. This system will facilitate understanding of neural mechanisms underlying behavioral responses to mechanical stimuli.

Calcium Imaging in Freely Behaving Caenorhabditis elegans with Well-Controlled, Nonlocalized Vibration

Reported here is a system for calcium imaging in freely behaving Caenorhabditis elegans with well-controlled, nonlocalized vibration. This system allows researchers to evoke nonlocalized vibrations with well-controlled properties at nano-scale displacement and to quantify calcium currents during responses of C. elegans to the vibrations.

自由行为 秀丽隐杆线虫 的钙成像，具有良好控制，非局部化振动

Nonlocalized mechanical forces, such as vibrations and acoustic waves, influence a wide variety of biological processes from development to homeostasis. ...

Investigating Pain-Related Avoidance Behavior using a Robotic Arm-Reaching Paradigm

Avoidance behavior is a key contributor to the transition from acute pain to chronic pain disability. Yet, there has been a lack of ecologically valid paradigms to experimentally investigate pain-related avoidance. To fill this gap, we developed a paradigm (the robotic arm-reaching paradigm) to investigate the mechanisms underlying the development of pain-related avoidance behavior. Existing avoidance paradigms (mostly in the context of anxiety research) have often operationalized avoidance as an experimenter-instructed, low-cost response, superimposed on stimuli associated with threat during a Pavlovian fear conditioning procedure. In contrast, the current method offers increased ecological validity in terms of instrumental learning (acquisition) of avoidance, and by adding a cost to the avoidance response. In the paradigm, participants perform arm-reaching movements from a starting point to a target using a robotic arm, and freely choose between three different movement trajectories to do so. The movement trajectories differ in probability of being paired with a painful electrocutaneous stimulus, and in required effort in terms of deviation and resistance. Specifically, the painful stimulus can be (partly) avoided at the cost of performing movements requiring increased effort. Avoidance behavior is operationalized as the maximal deviation from the shortest trajectory on each trial. In addition to explaining how the new paradigm can help understand the acquisition of avoidance, we describe adaptations of the robotic arm-reaching paradigm for (1) examining the spread of avoidance to other stimuli (generalization), (2) modeling clinical treatment in the lab (extinction of avoidance using response prevention), as well as (3) modeling relapse, and return of avoidance following extinction (spontaneous recovery). Given the increased ecological validity, and numerous possibilities for extensions and/or adaptations, the robotic arm-reaching paradigm offers a promising tool to facilitate the investigation of avoidance behavior and to further our understanding of its underlying processes.

Avoidance is central to chronic pain disability, yet adequate paradigms for examining pain-related avoidance are lacking. Therefore, we developed a paradigm that allows investigating how pain-related avoidance behavior is learned (acquisition), spreads to other stimuli (generalization), can be mitigated (extinction), and how it may subsequently re-emerge (spontaneous recovery).

使用机械臂接触范式调查疼痛相关避免行为

Avoidance behavior is a key contributor to the transition from acute pain to chronic pain disability. Yet, there has been a lack of ecologically valid ...

Iterative Development of an Innovative Smartphone-Based Dietary Assessment Tool: Traqq

To collect dietary intake data in a fast and reliable manner, a flexible and innovative smartphone application (app) called Traqq was developed (iOS/Android). This app can be used as a food record and&#160;24-h recall (or shorter recall periods). Different sampling schemes can be created on either prespecified or random days/times within a predetermined period for both methods, with&#160;push notifications to urge the participants to register their food intake. In case of non-response, notifications are automatically rescheduled to ensure complete data collection. For use as a food record, respondents can access the app and log their food intake throughout the day. Food records close automatically at the end of the day; recalls close after submission of the consumed items. The recall as well as the food record module provide access to an extensive food list based on the Dutch food composition database (FCDB), which can be accustomed to fit different research purposes. When selecting a food item, respondents are simultaneously prompted to insert portion size, i.e., in household measures (e.g., cups, spoons, glasses), standard portion sizes (e.g., small, medium, large), or weight in grams, and eating occasion/time of consumption. Portion size options can be adjusted, e.g., only entry in grams in case of a weighed food record or time of consumption instead of eating occasion). The app also includes a My Dishes function, which allows the respondent to create their own recipes or product combinations (e.g., a daily breakfast) and only report the total quantity consumed. Subsequently, the app accounts for yield and retention factors. The data are stored on a secure server. If desired, additional questions, i.e., in general or those related to specific food items or eating occasions can be incorporated. This paper describes the development of the system (app and backend), including expert evaluations and usability testing.

This article describes the protocol for the development of an innovative smartphone-based dietary assessment application Traqq, including expert evaluations and usability testing.

创新智能手机饮食评估工具的迭倍开发：Traqq

To collect dietary intake data in a fast and reliable manner, a flexible and innovative smartphone application (app) called Traqq was developed ...

使用主动位置回避测试了解小鼠的空间学习

The Active Place Avoidance (APA) Test, an Effective, Versatile and Repeatable Spatial Learning Task for Mice

Hippocampus-dependent spatial learning in rodents has been tested using a variety of methods. These include the Morris water maze (MWM), Y-maze, and novel object location (NOL) tasks. More recently, the active place avoidance (APA) task has been developed as an alternative to these more traditional approaches. In the APA task, mice must use spatial cues placed around a rotating arena to avoid a stationary shock zone. Due to the multiple parameters that can be adjusted, the APA task has been demonstrated to be a very versatile approach. It lends itself to being used longitudinally and repeatedly for the same cohort of mice. Here, we provide a detailed protocol to successfully conduct the APA task. We also highlight alternative APA approaches that can be used to examine different components of spatial learning. We describe the data collection and analysis processes. Critical steps during the APA task are discussed to increase the likelihood of successfully conducting the test. The APA task has several advantages over more traditional spatial navigation tests. It is appropriate to use with aged mice or those with disease phenotypes such as Alzheimer's disease. The complexity of the task can be easily altered, allowing a wide range of mouse strains to be tested. Further, the APA task is suitable for testing animals that have undergone surgery or experimental interventions that may have affected motor or neural function, such as stroke or traumatic brain injury.

作者聚焦：研究衰老对海马依赖性空间学习的影响

Here, we present a protocol for the active place avoidance test, a hippocampus-dependent spatial learning paradigm designed for rodents. Altering key parameters allows for re-testing of animals before and after treatments or over time.

主动位置回避 （APA） 测试，一种有效、多功能且可重复的小鼠空间学习任务

Hippocampus-dependent spatial learning in rodents has been tested using a variety of methods. These include the Morris water maze (MWM), Y-maze, and novel ...

<meta charset="utf8"></head><body>秀丽隐杆线虫中挥发性信息素反应的提取和定量

Volatile Sex Pheromone Extraction and Chemoattraction Assay in Caenorhabditis elegans

Chemical communication is vital in organismal health, reproduction, and overall well-being. Understanding the molecular pathways, neural processes, and computations governing these signals remains an active area of research. The nematode Caenorhabditis elegans provides a powerful model for studying these processes as it produces a volatile sex pheromone. This pheromone is synthesized by virgin females or sperm-depleted hermaphrodites and serves as an attractant for males.
This protocol describes a detailed method for isolating the volatile sex pheromone from several C. elegans strains (WT strain N2, daf-22, and fog-2) and C. remanei. We also provide a protocol for quantifying the male chemotaxis response to the volatile sex pheromone. Our analysis utilizes measurements such as chemotaxis index (C.I.), arrival time (A.T.), and a trajectory plot to compare male responses under various conditions accurately. This method allows for robust comparisons between males of different genetic backgrounds or developmental stages. Furthermore, the experimental setup outlined here is adaptable to investigating other chemoattraction chemicals.

<meta charset="utf8"></head><body>作者聚焦：检查挥发性信息素对雄性 秀丽隐杆线 虫行为的影响

This protocol establishes methods for extracting and quantifying responses to the volatile sex pheromone in C. elegans, providing tools to study chemical communication and navigation trajectory.

秀丽隐杆线虫中的挥发性信息素提取和趋化试验

Chemical communication is vital in organismal health, reproduction, and overall well-being. Understanding the molecular pathways, neural processes, and ...

一种基于智能手机的秀 丽隐杆线虫 草坪回避测定成像方法

一种基于智能手机的秀丽隐杆线虫草坪回避测定成像方法