Alfredo Attisano was supported by a European Social Fund studentship. James T. Murphy is supported by USDA-NIFA Award 2013-34103-21437.

1.构建飞行磨<ol><li> 构建亚克力塑料支撑结构 ： <ol><li>切成3毫米厚的透明压克力板成两个外垂直壁中，一个中央垂直壁和所指定的图 1中所示的设计的五个水平架。 </li><li>成垂直壁（ 图1和2; OW和CW），以形成所述支撑结构（图2A）;通过插入所述货架（HS 图1和图2）装配。 </li><li>通过在外角插入聚苯乙烯列在该装置（图2A和图2C）的背面加强的结构。如果需要的话，胶短件直角边缘保护器沿中心垂直壁结到用于水平搁板提供额外的支撑。 </li></ol></li><li> 构建pivotin摹臂总成： <ol><li>胶的1厘米直径的塑料管5厘米长成每个电池单元的顶部中心。胶的1厘米直径的塑料管有2厘米长到每个细胞的底部中心，并确保每个单元中的顶部和底部管道对准。用热胶，贴上两个10毫米×4毫米N42钕磁铁给每个支持的结束，形成磁轴承工厂的手臂。 </li><li>插入一个昆虫针到20微升的吸头和到位用热胶固定。定位销，使得两个端延伸出来的枪头，以形成飞行轧机的电枢。 注意：在飞行试验中，所述销的顶部被代替由顶组磁体保持。底组磁体是保持衔铁处于垂直位置，使其能够围绕其轴线旋转。 </li><li>切的19计非磁性皮下钢管24厘米长。用热胶，贴上中心点枪头f的顶部ROM步1.2.2。管在2厘米从端至95°的角度，而使从中心点12cm的长臂和短臂从中心向弯曲（图2B）10厘米半径的弯曲的一端。 注意：半径长度可以适应不同的革命的距离而改变。 </li></ol></li><li> 设置红外传感器和数据记录器 ： <ol><li>固定的红外线传感器使用可重复使用的粘合剂腻子每个电池单元的永恒两侧，允许传感器通过切入外部垂直壁支撑件（ 图2C）中的开口延伸到细胞中。 </li><li>通过建立在一个焊试验电路板（图3）一个非常基本的电子电路连接的红外线传感器到数据记录器。在实验电路板（ 图3A，B），红外连接的输入和输出连接180Ω和2.2kΩ的两个电阻分别。发生在阿尔特电阻沿着线路板到来自多个传感器的记录期间减少在电压信号滴rnate行（参见图3C）。 </li></ol></li></ol> 2.飞行试验<ol><li> 间接通过昆虫针系绳昆虫飞行磨臂 ： <ol><li>放置一个小箔标志在枢转臂的未弯曲端的端部以最大化在传感器中的红外线光束的中断和充当平衡重。 </li><li>根据昆虫的大小和角质层区域可用于连接，连接实验昆虫昆虫引脚可重复使用的粘合剂腻子或无毒的皮胶。如果有必要，无论是冷冻或用二氧化碳麻醉昆虫。 </li><li>塑造少量的昆虫学销的圆形尖端周围粘合剂腻子并覆盖它用一滴无毒皮胶。轻轻涂抹于前胸背板区，等待5-10s，直到胶水变干。 注：亲cedure步骤2.1.3适合于昆虫坚硬（甲虫，臭虫）或软（黄蜂，苍蝇）角质层。昆虫与毛茸茸的角质层（蛾，蝶）将需要与圈养之前很细的画笔轻轻去除毛发。 </li><li>插入附加到枢转臂组件的弯曲端的昆虫的管脚。 </li><li>试飞结束后，取出栓系用细钳子。 注意：数据记录器设置和获取已被优化如下在材料表中列出的具体的设备和应调整为与另一种设备用。 </li></ol></li><li> 启动与免费提供WinDAQ精简版软件记录会话 <ol><li>下载并安装免费软件WinDAQ精简版（见设备清单）。 </li><li>打开仪器硬件管理器中，选择数据记录器，从弹出的列表，然后按“开始Windaq”。一个新的窗口将打开，输入标志人从每个传感器将被显示。 </li><li>选择所需要的采样频率在该数据记录器中读取并显示传感器的输出。 注意：采样频率将取决于昆虫的飞行速度，但是采样频率30-45赫兹之间的范围内将是足够快，以捕捉中小规模的昆虫的飞行。 </li><li>按Ctrl-F4开始记录会话。选择第一个弹出窗口中的录音文件的目标路径。选择一段适当的长度，记录飞行的特定昆虫和实验。定义录音时间，在第二个弹出式窗口。一旦录音时间结束后按Ctrl-S来完成录制的文件。 </li></ol></li><li> 检查录音质量。 <ol><li>打开记录的飞行轨迹，并选择一个电压通道。按Ctrl-T与电压的统计数据为每个通道打开一个弹出窗口。 </li><li>确保没有大滴在最低值导致从整个电路（图4）的电压降。丢弃在其中信道平均和最小电压之间的差大于0.1V。任何频道</li></ol></li><li> 将文件保存在一个* .csv格式 ：选择File&gt;另存为，然后在弹出的窗口中选择“电子表格打印（CSV）”。在“电子表格注释”，弹出的窗口中选择“相对时间”，并取消选择所有其他选项。点击确定保存文件。 </li></ol>飞行数据的使用Python 3.4.x 3.分析<ol><li> 安装最新的Python 3.4.x版本 ，下载档案<a href="https://www.jove.com/files/ftp_upload/53377/Python_scripts.zip">Python_scripts.zip（</a>补充文件），打开它，并保存standardize_peaks.py和flight_analysis.py到桌面上。 </li><li> 规范和选择记录信号的峰如下<ol><li>在standardize_peaks.py图标上单击右键。选择“打开方式IDLE”。 注：空闲的默认编辑器为Python，但任何文本编辑器，可用于这一目的。 </li><li>在线路18-19，指定周围用来执行为每个信道的电压信号的标准化的平均电压的阈值。 注意：默认值被设置为提供一个微调信号标准化，但用户可以根据平均电压为每个信道的值定义任何所需的阈值。这些都可以在电压统计窗口中找到（见步骤2.3）。 </li><li>在第45行，键入路径，其中记录的* .csv文件被保存的文件夹。 </li><li>在91行，键入路径到要在其中记录* .TXT峰值文件的文件夹。 </li><li>在管线61和管线72中，指定所需要的信道数。添加或删除＃在行61-63和72-74的开始到一个最大删除频道imum的8个通道。 </li><li>保存文件，然后按F5键启动脚本。 </li><li>在该弹出窗口，然后按返回输入* .csv文件（以及任何额外的子文件夹）的名称保存一个新的* .TXT文件中指定的文件夹中的标准化信号。 注意：根据使用的n个信道的数量，该文件包含n + 1个列：第一列是取样事件的相对时间，其他的n列表示从用于记录的n个通道的基极和峰事件。值0表示的基极电压，而值1表示从标志通过IR传感器的通路的峰。 </li></ol></li><li> 使用标准化文件分析航迹 ：编辑flight_analysis.py脚本，以适应用户的实验条件： <ol><li>右键单击flight_analysis.py图标。选择“打开方式IDLE”。 </li><li>在线路39和线路80调整长度根据臂的半径的圆形的飞行路径。 </li><li>如果需要，通过删除＃中的行50-52激活可选的速度校正循环。相应地更改速度值。 </li><li>在管线77和管线85，编辑速度阈值和时间间隔值来校正在飞行轨道假速度读数和占两个连续长时间不间断飞行较量之间发生非常短的时间间隙。 </li><li>在198行中，指定以秒为单位的总记录时间。改变从线287起，输出线的数值范围。 注意：默认范围可以根据用户的实验要求进行修改。为了做到这一点，在该函数内的所有数值（包括那些在变量名，例如在变量“flight_300_900”）需要被改变成所需的值。 </li><li>在线路248类型的文件夹路径，其中，* .TXT标准化文件被保存。 </li><li>指定号码渠道。添加或通过在线257-259，270-272线条和线条279-281最多8个信道的开头添加或删除＃删除频道。 </li><li>在304行中键入路径，要在其中保存输出文件的文件夹。 </li><li>一旦指定用户的所有设置，保存文件，然后按F5键启动脚本。 </li><li>输入* .TXT文件的名称来分析（以及任何额外的子文件夹），在弹出的窗口中，然后按返回。 </li></ol></li></ol>

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The authors have nothing to disclose.

经济实惠，灵活，可调的飞行工厂设计 。 昆虫飞行的行为感兴趣的是一系列的科学家，从那些在变量的环境中专家生物防治谁需要了解的条件如何影响的害虫驱散倾向感兴趣的昆虫的基本行为。飞行行为，可以通过其范围从飞行“跑步机”和风洞是近似的现场条件，以静态的系留飞行装置的各种方法进行研究。系留飞行轧机，像一个这里提出，在飞行的某些方面，例如高度变化的限制，不能测量14。然而，系留飞行造纸厂并允许昆虫飞不间断，从而使研究人员量化参数，如飞行速度，距离和周期性和这些参数与环境条件的，生理学和米关联orphology。 飞行磨这里介绍的目的是让研究人员没有电子建造和使用系留飞行磨为了研究昆虫的飞行行为的专门知识。这种设计的一个优点是，与其它设计相比飞行轧机的总成本是低的。总成本可保持300远低于美元。塑料压克力板是最昂贵的项目。第二个优点是，该飞行轧机可适于在许多实验室中可用的有限控制条件的工作区，而不是一个专门风洞。采用3毫米厚的透明的丙烯酸塑料片意味着该结构既透明的，以允许容易观察昆虫的，并且还具有重量轻，从而使飞行轧机被移动到适当的位置用于飞行试验。飞行磨单元的堆叠配置最大化，同时尽量减少脚的样本数执行打印装置的。另外，该装置可容易地拆卸用于存储。此外，飞行磨机被设计为允许对大量个人可以相对容易地采样。每次飞行磨包含8个细胞，使研究人员能够同时记录多个个体的飞行活动。通过昆虫销间接附着昆虫的枢转臂允许个别昆虫被迅速地放在并除去从飞行轧机。最后，数据记录电子是简单，易于使用，具有进行数据分析可免费获得的软件。一旦组装好，飞行磨机使用简单的IR传感器来记录飞行活动。箔标志在通过红外光束的臂的端部的通道允许被记录的臂的每一转。旋转速率可以像速度数据，行驶距离，总飞行时间和飞行模式，以被记录为输入到一个数据记录器。 飞行磨能够适于许多不同种类的昆虫的。因为，即使重，所产生的阻力由窄直径减小，允许甚至小昆虫是飞行试验的枢转臂的使用皮下注射钢管比其它选项，如木棍或吸管更有效。最近，小件光纤已被用于在飞行磨小昆虫25。臂的弯曲结局可以粘到衔铁在相对于支撑轴不同的角度，以在其天然飞行方向定位实验昆虫。在提出的设计，其中，半径为10厘米的长度，在一个旋转行进的整个距离是62.8厘米。除去中央纵壁将允许飞行轧机，其中所述臂的半径的长度可以增加一倍，以容纳较大的昆虫和公转距离可达1.20米的另一种结构。在这种情况下，强磁体是recommenDED，以适应和稳定再磨的手臂。 如所陈述在整个的飞行工厂设计的灵活性和适应性为感兴趣的昆虫物种和研究人员能够定制他们的特殊需要。这包括昆虫，不仅身体的需要，包括参数，如尺寸，功率，角质层的结构，而且物种之间的生物学差异。一个潜在的缺点，以所有飞行钢厂是缺乏跗支持“分裂势力”昆虫飞行，也许是为了耗尽。虽然这是事实在一些物种中，例如，我们观察到的自动飞行的反应与我们的马利筋错误的试验，它是不是适用于所有我们已经测试（例如N. vespilloides）的昆虫。但是，即使有自动响应，我们从来没有观察昆虫飞行劳累或死亡，部分是因为录制时间，我们选择以适应昆虫的生物学。因此，为了做到这一点是很重要在感兴趣的昆虫初步观察，以了解，为了优化数据采集其在飞行磨机行为。另外一个，著名的问题，飞行厂，是惯性能保持甚至在昆虫已经停止积极飞行运动。该脚本提供了占误读由于飞行磨的惯性，在飞行速度特点是快速减少和增加峰之间的距离。该脚本flight_analysis.py“放弃这些”假峰“，并构造分析的新信号。用户可以选择校正的速度阈值，因为在脚本中提供的说明解释。 5 V的电源是足以获得一个可读电压信号，但是具有可变输出电压的电源装置可被用作动力源，以允许输入功率是变化的，从而优化工作电压为每个传感器。这种解决方案还可以帮助提高可视化的质量Ø˚F高峰信号在软件的录音界面。传感器的输出显示在软件接口由碱和峰值电压，其中基准电压表示来自传感器在休息的最低输出电压（当红外线光束不被中断），而峰值电压是从基极电压的上升所形成当作为手臂穿过光束红外光束被中断时才会发生。 5伏的输入电压提供了大约100 mV的上升而增加输入到7V增加了峰值的上升到300mV，允许为基础，峰值电压更清晰的歧视。所选择的焊面包板的大小决定了许多飞行细胞如何能得到满足。为了从多个传感器在记录期间最小化在电压信号下降时，建议以放置在沿着线路板交替行的电阻器（参见图3C）。 可定制的信号标准化和analysi对开放存取的编程语言Python的s要写的脚本。 电压信号的标准化和分析，通过使用Python中，这是一个免费的，广泛使用的通用型，高层次的编程语言定制编写的脚本进行的。最终用户可以轻松地自定义脚本与自己指定的设置工作。定制是通过简单地改变数值或变量名来实现。关于如何自定义参数的注意事项可以在脚本本身中找到。脚本中的默认值设定为提供一个微调信号标准化，但用户可以根据平均电压为每个信道的值定义任何所需的阈值。在飞行分析剧本，从线105功能flying_bouts计算持续时间最长秒，最短的飞行较量，时间在飞行花了的总记录时间的百分比和飞来飞去事件的数量s的指定时间范围内。该范围可以根据用户的实验要求进行修改。为了做到这一点，在该函数内的所有数值（包括那些在变量名，例如在变量“flight_300_900”）需要被改变成所需的值。范围和它们的持续时间的数目简单地取决于用户的规格。该脚本将打印在屏幕上的分析对每个信道的结果。这些措施包括：平均飞行速度，总飞行时间，行驶距离，最短和最长飞行较量和飞行组成。此外，脚本返回* .DAT文件为每个信道，并将其保存在由用户指定的输出文件夹。每* .DAT文件包含两列：第一个代表峰值事件的相对时间，第二次是两个连续峰值之间的事件的详细速度变化。此文件可以在Excel或R导入到生产速度变化的图表里时间和可视化的飞行活动模式。 总之，这些结果表明，该飞机厂的设计可以很容易地和成功地实施，以聚集行为研究观察飞行活动模式在不同的昆虫模型的数据。这样的数据可以被用来研究在运动模式那样依赖例如在生理学和形态学的个体差异。这提供了极大分析上市公司潜在的生理和形态特征确定的运动模式，如觅食或迁徙活动，最终影响整个人口的个体差异。随着时间的推移详细速度变化可与详细的生理学和形态学测量组合使用，将提供来研究资源消费模式或变化的飞行活动身体部分的形态的效果的工具。

几个实验室技术已经开发了用于昆虫飞行行为1,2-研究。这些范围从简单的静态圈养3,4，使运动的栓昆虫5更大的自由先进的设备。迄今为止飞行室6-9所代表的设备允许的飞行在受控条件下的自由的最高水平。这种技术具有两个主要的缺点：它是难以用于大型的昆虫的研究和数据收集的手动过程是耗时的。 飞行钢厂表示为昆虫飞行实验室条件下10月12日的研究中最常见和负担得起的技术之一。这种技术是最好的静态束缚，因为它提供了运动刺激的13，但它不同于自由飞行的行为反应14-16。上的轧机和在野外的飞行特性的某些方面similař5,17所以尽管有一些限制，飞行钢厂表示一个可行的选择，调查关于特定飞行行为应答的发生的问题，由于是迁徙飞行型的情况下。此外，飞行钢厂更容易实现比风洞或飞行室和数据收集可以很容易地实现自动化。因此，研究人员感兴趣的飞行行为常常会发现飞行钢厂是最好的选择，但应该意识到该方法的潜在限制。在这里，一个灵活，可定制的飞行工厂设计，提出对于那些已经选择了利用飞行钢厂进行调查飞行行为的研究。 几位作者描述的替代航班轧机的设计。在飞行轧机系统， 即 ，枢转磨机的臂的一般的主要部分，是非常简单的实现。少是直截了当的飞行轧机系统，它允许数据的记录的电子部件。与埃尔处理ectronic电路的设计可以是具有挑战性的，特别是对于昆虫学家或行为生态学家缺乏电子的背景知识。有些作者描述了他们的飞行工厂设计18-21，或飞行机的电子部件的缺失22,23说明一个复杂的或过时的电子电路元件。其他设计描述结构复杂actographs，这是相当复杂的实现，但可以帮助研究人员进行更复杂的行为观察5。 本文设计了一个简单的打造，价格相对低廉的飞行磨的系留飞行昆虫的研究描述。连同极其简单的电子部件，该设计具有许多优点。飞行轧机设计在通常在标准昆虫生态实验室中可用的约束空间中使用。其结构是做出来的透明亚克力的P拉斯蒂克使得单个光源可以均匀地达到在该轧机的单独的腔室每一个人。给定的物质和小尺寸的透明度，飞行磨机可以在培养箱中标准化的光照和温度条件下使用。最后，整个结构可以组装和拆卸容易，一旦拆卸，它可以储存在一个小的空间。另一个优点与结构的设计是，飞行轧机可以被修改，以允许不同大小的昆虫，并使用不同的公转距离的研究。这次飞行磨已被用于收集昆虫等不同大小的数据和形状马利筋错误，Oncopeltus暗纹东方鲀 24，葛根错误， 筛豆龟蝽和埋葬甲虫，Nicrophorus vespilloides。飞行工厂设计还允许通过量高的要求，需要大样本量的研究。数据可以使用8个通道同时为每个数据记录器ü收集SED使大量的个体可以同时分析和大量的样品可以在同一天进行处理。不需要昂贵的软件是必要的记录和可视化的数据和用于数据分析的定制编写脚本可以下列实验设计的具体需要来修改。逃反应是在不同的昆虫种类变化很大。因此，进行全面的飞行磨实验前，建议对焦点昆虫模型的飞行响应初步测试。这些将提供行为变化在飞行的反应，这将用于在飞行分析微调方面，例如记录时间或飞行速度范围的程度的理解。

<table border="0" cellpadding="0" cellspacing="0" width="854">
	<tbody>
		<tr height="147">
			<td height="147" style="height:147px;width:248px;">Data Logger</td>
			<td style="width:137px;">DATAQ Instruments, Ohio, USA</td>
			<td style="width:137px;">DI-149</td>
			<td style="width:332px;">These particular data loggers were chosen because they can be easily connected via USB to a computer and come with free proprietary software (WinDaq/Lite, DATAQ Instruments, Ohio, USA) to visualize and record the sensor&#39;s output, increasing the affordability of the flight mill design.</td>
		</tr>
		<tr height="126">
			<td height="126" style="height:126px;width:248px;">Data Logger - potential alternative</td>
			<td style="width:137px;"></td>
			<td style="width:137px;"></td>
			<td style="width:332px;">A potential alternative to the DATAQ data loggers&#160; is an RS232 to USB adaptor, readily available through office or electronic supply stores.&#160; These should be able to read data directly from the serial port via the pyserial module.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:248px;">Entomological pins</td>
			<td style="width:137px;">BioQuip</td>
			<td style="width:137px;"></td>
			<td style="width:332px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:248px;">Hypodermic steel tubing 19 guage</td>
			<td style="width:137px;">Small Parts</td>
			<td style="width:137px;">B000FN5Q3I</td>
			<td style="width:332px;">Available through Amazon.com; other suppliers are available but be sure to purchase austenitic steel tubing to ensure the arm in non-magnetic</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:248px;">IR Sensors</td>
			<td style="width:137px;">Optek Technology Inc., Texas USA</td>
			<td style="width:137px;">OPB800W</td>
			<td style="width:332px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:248px;">N42 neodymium magnets</td>
			<td style="width:137px;"></td>
			<td style="width:137px;"></td>
			<td style="width:332px;">Readily available; can be purchased through specialized magnet suppliers, hobby stores or Amazon</td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:248px;">Plexiglass/perspex</td>
			<td style="width:137px;"></td>
			<td style="width:137px;"></td>
			<td style="width:332px;">Readily available at any hardware store</td>
		</tr>
		<tr height="105">
			<td height="105" style="height:105px;width:248px;">Polystyrene columns for support</td>
			<td></td>
			<td></td>
			<td style="width:332px;">Any polystyrene or styrofoam packing materials that might otherwise be discarded or recycled can be used to fashion the support columns for the flight mill.&#160; Otherwise, styrofoam insulation sheets are available at any hardware store.</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:248px;">Solderless Breadboard Power Supply Module</td>
			<td style="width:137px;">Arrela</td>
			<td style="width:137px;">MB102</td>
			<td style="width:332px;">The 5V power unit, breadboard and solderless male-male jumper wires can be easily purchased as a kit.</td>
		</tr>
	</tbody>
</table>

a simple flight mill for the study of tethered flight in insects

Flight in insects can be long-range migratory flights, intermediate-range dispersal flights, or short-range host-seeking flights. Previous studies have shown that flight mills are valuable tools for the experimental study of insect flight behavior, allowing researchers to examine how factors such as age, host plants, or population source can influence an insects' propensity to disperse. Flight mills allow researchers to measure components of flight such as speed and distance flown. Lack of detailed information about how to build such a device can make their construction appear to be prohibitively complex. We present a simple and relatively inexpensive flight mill for the study of tethered flight in insects. Experimental insects can be tethered with non-toxic adhesives and revolve around an axis by means of a very low friction magnetic bearing. The mill is designed for the study of flight in controlled conditions as it can be used inside an incubator or environmental chamber. The strongest points are the very simple electronic circuitry, the design that allows sixteen insects to fly simultaneously allowing the collection and analysis of a large number of samples in a short time and the potential to use the device in a very limited workspace. This design is extremely flexible, and we have adjusted the mill to accommodate different species of insects of various sizes.

图5示出了可以使用在上一节中所描述的脚本来获得图形的类型的代表性例子。飞行数据来自使用埋葬甲虫Nicrophorus vespilloides为模型（Attisano， 未发表的数据）的动物学系在剑桥大学进行的试点工作获得的。约20日龄两个年轻未交配的雄性都拴飞行钢厂和摆在14时10大号控制的环境条件：D和21℃。甲虫被留在飞行轧机连续8小时，并在飞行活动的记录。屏幕上的分析和图形输出使得有可能解决在飞行中的活动模式的个体差异。例如，第一公（图5A）表现出强烈的飞行活动记录的第一个小时之内，其特征在于，高速和连续飞行，历时约3小时。钍被延长活性相从约1.6米/秒的特征在于在速度逐渐降​​低到约1米/秒其中。最初的飞行回合后，个人表现出相对较短的飞行几乎周期模式的较量每次约10-15分钟时间。第二阳显示出非常不同的飞行模式与飞行较量从未超过15-20分钟（图5B）的持续时间。在此个人飞行活动的特征在于一个宽的飞行较量的传播中的前4个小时的记录，之后，其活性变得几乎周期性。这个人也提出了非常低的飞行速度，只有偶尔超过0.4米/秒。 使用不同的昆虫模型，马利筋错误Oncopeltus鲀得到另一个有代表性的例子。在迁徙行为和马利筋错误女性24生理反应的食物应力研究过程中收集的数据。在这项研究中的录音时间设定为一小时，以刻画女性为移民或居民。这些行为类型的特征在于“全有或全无”的响应。迁徙女性从事持续不断航班通常持续几个小时，而女性居民从来没有表现出飞行活动超过几分钟。因此，农民女性将呈现飞行图案像在图6A中，而居民女性的特点将是像在图6B的运动模式。 <img alt="图1" src="/files/ftp_upload/53377/53377fig1.jpg" /> 丙烯酸塑料支撑结构图1.设计配置。的丙烯酸塑料支撑结构为飞行轧机是从三个不同的部件构成。有含有插槽的货架和一个开口，以容纳红外SENS两头在外垂直壁（OW）口服补液盐（A）。有一个单一的中央纵壁（CW）与货架槽。并且有5个水平货架（HS）与墙壁插槽。磁轴是粘在水平货架上的位置B. <a href="https://www.jove.com/files/ftp_upload/53377/53377fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/53377/53377fig2.jpg" /> 图2.组装飞机厂。 （A）中的丙烯酸塑料支撑结构通过滑动在两个外壁（OW）和中央壁（CW）的五个水平搁板（HS）到槽，所获得的结构具有8个独立的细胞各包含磁力组装枢轴和IR传感器，允许8个人在同一时间飞行。 （B）中的枢轴臂到的昆虫被拴系可构造成accomm<html>大馆各种尺寸和昆虫的形貌。 （三）由于拴昆虫移动转臂悬挂磁铁之间，箔标志在手臂的另一端激活红外传感器（箭头）。 <a href="https://www.jove.com/files/ftp_upload/53377/53377fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/53377/53377fig3.jpg" /> 图3.设计中的红外传感器连接到数据记录器的电路 。 （一）一个简单的电路由红外传感器数据记录器连接的输入。 （B）中的每个数据记录器可以被供电并经由使用图一焊试验电路板连接到数据记录器。 （C）的多个传感器可以连接到使用相同的试验板的单个数据记录器。large.jpg“目标=”_空白“&gt;点击此处查看该图的放大版本。 <img alt="图4" src="/files/ftp_upload/53377/53377fig4.jpg" /> 图中记录的飞行活动4.示例。电压峰代表飞行工厂的手臂彻底革命。 （A）飞行事件的高品质录音，无压降所记录的信号。 （B）的飞行事件，在记录信号的电压降。 <a href="https://www.jove.com/files/ftp_upload/53377/53377fig4large.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/53377/53377fig5.jpg" /> 图从埋葬甲虫Nicrophorus vespilloides代表性的飞行数据 。在飞行中的个体变异行为很容易在飞行记录确认。 （ 一）持续一个单独飞行了大约三个小时的庭审开始后，然后在整个审判的其余部分周期性地扑到高速。 （ 二）个人的行为是不同的这种甲虫飞只是偶尔在整个审判，永远飞在看到在A组（注意在Y轴上规模的差异）个人高的速度。 <a href="https://www.jove.com/files/ftp_upload/53377/53377fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图6" src="/files/ftp_upload/53377/53377fig6.jpg" /> 图从乳草错误Oncopeltus鲀代表性的飞行数据 。行为的两个不同的图案的飞行数据记录之间进行清楚地观察。 （A </strong&gt;）这种记录是典型的飞行特性见于洄游个体的类型。迁徙个人在一个相对稳定的速度在长时间飞行。 （二）在A组的行为对比与居民个人的典型飞行的行为。居民在较低的速度和飞行较量飞行只持续很短的时间（注意A和B的X轴在规模上的差异）。 <a href="https://www.jove.com/files/ftp_upload/53377/53377fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about A Simple Flight Mill for the Study of Tethered Flight in Insects at JoVE.com

A Simple Flight Mill for the Study of Tethered Flight in Insects

Flight in insects is influenced by a number of factors and the propensity to disperse is an important variable in understanding insect ecology and biological control strategies. We describe the construction and use of a simple, relatively inexpensive, and flexible flight mill for measuring parameters of tethered flight in insects.

一个简单的飞行磨的系留飞行的昆虫研究

Flight in insects can be long-range migratory flights, intermediate-range dispersal flights, or short-range host-seeking flights. Previous studies have ...

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Research

JoVE Journal

Neuroscience

16.8K 次观看.  Polish Academy of Sciences. Flight in insects is influenced by a number of factors and the propensity to disperse is an important variable in understanding insect ecology and biological control strategies. We describe the construction and use of a simple, relatively inexpensive, and flexible flight mill for measuring parameters of tethered flight in insects.

视频: A Simple Flight Mill for the Study of Tethered Flight in Insects

A Simple Composite Phenotype Scoring System for Evaluating Mouse Models of Cerebellar Ataxia

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebella ataxia. It is derived from previously published phenotype assessments in several disease models, including spinocerebellar ataxias, Huntington s disease and spinobulbar muscular atrophy. Measures include hind limb clasping, ledge test, gait and kyphosis. Each measure is recorded on a scale of 0-3, with a combined total of 0-12 for all four measures. The results effectively discriminate between affected and non-affected individuals, while also quantifying the temporal progression of neurodegenerative disease phenotypes. Measures may be analyzed individually or combined into a composite phenotype score for greater statistical power. The ideal combination of the four described measures will depend upon the disorder in question. We present an example of the protocol used to assess disease severity in a transgenic mouse model of spinocerebellar ataxia type 7 (SCA7).
Albert R. La Spada and Gwenn A. Garden contributed to this manuscript equally.

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebellar ataxia. Measures include hind limb clasping, ledge test, gait and kyphosis. This protocol effectively discriminates between affected and non-affected individuals, and detects the progression of affected individuals over time.

一个简单的综合评分系统评估小脑共济失调的小鼠模型的表型

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebella ataxia. It is derived from previously ...

科研

神经科学

The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice

Previous characterizations of rodent eating behavior have revealed that they use coordinated forepaw movements to manipulate food pieces. We have extended upon this work to develop a simple quantitative measure of forepaw dexterity that is sensitive to lateralized impairments and age-dependent changes. Rodents learn skillful forepaw and digit movements to manage thin pasta pieces, which they eagerly consume. We have previously described methods for quantifying vermicelli handling in rats and showed that the measures are very sensitive to forelimb impairments resulting from unilateral ischemic lesions, middle cerebral artery occlusions and unilateral striatal dopamine depletion [Allred, R.P., Adkins, D.L., Woodlee, M.T., Husbands, L.C., Maldonado M.A., Kane, J.R., Schallert, T. &amp; Jones, T.A. The Vermicelli Handling Test: a simple quantitative measure of dexterous forepaw function in rats. J. Neurosci. Methods 170, 229-244 (2008)]. Here we present a more detailed protocol for this test in rats and compare it with a newly developed version for mice, the Capellini Handling Test. Rats and mice are videotaped while handling short lengths of uncooked vermicelli or capellini pasta, respectively, with a camera positioned to optimize the view of paw movements. Slow motion video playback allows for the identification of forepaw adjustments, defined as any distinct removal and replacement of the paw, or of any number of digits, on the pasta piece after eating commences. Forepaw adjustments per piece are averaged over trials per each testing session. Repeated testing permits sensitive quantitative analysis of changes in forepaw dexterity over time. Protocols for pre-testing habituation and handling practice, as well as procedures for characterizing atypical handling patterns, are described. Because rats and mice perform the pasta handling tests slightly differently, species-specific differences in administration and scoring of these tests are highlighted. All animal use was in accordance with protocols approved by the University of Texas at Austin Animal Care and Use Committee.

The Vermicelli and Capellini Handling Tests of forepaw dexterity take advantage of the natural inclination of rodents to manipulate food items using skillful forepaw and digit movements. Animals are videotaped while handling short strands of uncooked dry pasta. Slow motion video playback allows for the quantification of forepaw adjustments.

粉条和Capellini处理测试：在大鼠和小鼠的灵巧的前爪功能简单的定量措施

Previous characterizations of rodent eating behavior have revealed that they use coordinated forepaw movements to manipulate food pieces. We have extended ...

Studying the Neural Basis of Adaptive Locomotor Behavior in Insects

Studying the neural basis of walking behavior, one often faces the problem that it is hard to separate the neuronally produced stepping output from those leg movements that result from passive forces and interactions with other legs through the common contact with the substrate. If we want to understand, which part of a given movement is produced by nervous system motor output, kinematic analysis of stepping movements, therefore, needs to be complemented with electrophysiological recordings of motor activity. The recording of neuronal or muscular activity in a behaving animal is often limited by the electrophysiological equipment which can constrain the animal in its ability to move with as many degrees of freedom as possible. This can either be avoided by using implantable electrodes and then having the animal move on a long tether (i.e. Clarac et al., 1987; Duch &amp; Pfl&uuml;ger, 1995; B&ouml;hm et al., 1997; Gruhn &amp; Rathmayer, 2002) or by transmitting the data using telemetric devices (Kutsch et al, 1993; Fischer et al., 1996; Tsuchida et al. 2004; Hama et al., 2007; Wang et al., 2008). Both of these elegant methods, which are successfully used in larger arthropods, often prove difficult to apply in smaller walking insects which either easily get entangled in the long tether or are hindered by the weight of the telemetric device and its batteries. In addition, in all these cases, it is still impossible to distinguish between the purely neuronal basis of locomotion and the effects exerted by mechanical coupling between the walking legs through the substrate. One solution for this problem is to conduct the experiments in a tethered animal that is free to walk in place and that is locally suspended, for example over a slippery surface, which effectively removes most ground contact mechanics. This has been used to study escape responses (Camhi and Nolen, 1981; Camhi and Levy, 1988), turning (Tryba and Ritzman, 2000a,b; Gruhn et al., 2009a), backward walking (Graham and Epstein, 1985) or changes in velocity (Gruhn et al., 2009b) and it allows the experimenter easily to combine intra- and extracellular physiology with kinematic analyses (Gruhn et al., 2006).
We use a slippery surface setup to investigate the timing of leg muscles in the behaving stick insect with respect to touch-down and lift-off under different behavioral paradigms such as straight forward and curved walking in intact and reduced preparations.

We describe a method to record motor activity, timed to the electrically recorded tarsal contact signal in a tethered insect, walking on a slippery surface. This is used to study the neural basis of adaptive behavior under reduced influence of mechanical interaction between legs through the substrate.

研究昆虫的自适应运动能力行为的神经基础

Studying the neural basis of walking behavior, one often faces the problem that it is hard to separate the neuronally produced stepping output from those ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy and cheap methods to distinguish between individuals in a trial. A standard technique for this is CO2 anaesthesia and then labelling or wing clipping each fly. However, this is invasive and has been shown to affect behavior. Other techniques have used coloration to identify flies. This article presents a simple and non-invasive method for labelling Drosophila that allows them to be individually identified within experiments, using food coloring. This method is used in trials where two males compete to mate with a female. Dyeing allowed quick and easy identification. There was, however, some difference in the strength of the coloration across the three species tested. Data is presented showing the dye has a lower impact on mating behavior than CO2 in Drosophila melanogaster. The impact of CO2 anaesthesia is shown to depend on the species of Drosophila, with D. pseudoobscura and D. subobscura showing no impact, whereas D. melanogaster males had reduced mating success. The dye method presented is applicable to a wide range of experimental designs.

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

This protocol describes a simple, cost effective way to individually identify Drosophila or other insects. Demonstration data investigating mating success across three species of Drosophila show that this method is comparable or better than the use of CO2 anaesthesia.

染色昆虫行为检测：麻醉的交配行为<em&gt;果蝇</em

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy ...

In-depth Physiological Analysis of Defined Cell Populations in Acute Tissue Slices of the Mouse Vomeronasal Organ

In most mammals, the vomeronasal organ (VNO) is a chemosensory structure that detects both hetero- and conspecific social cues. Vomeronasal sensory neurons (VSNs) express a specific type of G protein-coupled receptor (GPCR) from at least three different chemoreceptor gene families allowing sensitive and specific detection of chemosensory cues. These families comprise the V1r and V2r gene families as well as the formyl peptide receptor (FPR)-related sequence (Fpr-rs) family of putative chemoreceptor genes. In order to understand the physiology of vomeronasal receptor-ligand interactions and downstream signaling, it is essential to identify the biophysical properties inherent to each specific class of VSNs.
The physiological approach described here allows identification and in-depth analysis of a defined population of sensory neurons using a transgenic mouse line (Fpr-rs3-i-Venus). The use of this protocol, however, is not restricted to this specific line and thus can easily be extended to other genetically modified lines or wild type animals.

Here, we describe a physiological approach that allows identification and in-depth analysis of a defined population of sensory neurons in acute coronal tissue slices of the mouse vomeronasal organ using whole-cell patch-clamp recordings.

在小鼠犁鼻急性组织切片定义细胞群的深入生理学分析

In most mammals, the vomeronasal organ (VNO) is a chemosensory structure that detects both hetero- and conspecific social cues. Vomeronasal sensory ...

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We demonstrate a simple way to tap into the fly reward system, modify experiences related to natural reward, and use voluntary ethanol consumption as a measure for changes in reward states. The approach serves as a relevant tool to study the neurons and genes that play a role in experience-mediated changes of internal state. The protocol is composed of two discrete parts: exposing the flies to rewarding and nonrewarding experiences, and assaying voluntary ethanol consumption as a measure of the motivation to obtain a drug reward. The two parts can be used independently to induce the modulation of experience as an initial step for further downstream assays or as an independent two-choice feeding assay, respectively. The protocol does not require a complicated setup and can therefore be applied in any laboratory with basic fly culture tools.

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for inducing rewarding and nonrewarding experiences in fruit flies (Drosophila melanogaster) using voluntary ethanol consumption as a measure for changes in reward states.

一个简单的方法来衡量在改建悬赏寻求行为使用<em&gt;果蝇</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the molecular and cellular basis of higher brain functions. A whole-mount preparation of adult brains with well-preserved morphology is critical for such whole brain-based studies, but can be technically challenging and time-consuming. This protocol describes an easy-to-learn, one-step dissection approach of an adult fly head in less than 10 s, while keeping the intact brain attached to the rest of the body to facilitate subsequent processing steps. The procedure helps remove most of the eye and tracheal tissues normally associated with the brain that can interfere with the later imaging step, and also places less demand on the quality of the dissecting forceps. Additionally, we describe a simple method that allows convenient flipping of the mounted brain samples on a coverslip, which is important for imaging both sides of the brains with similar signal intensity and quality. As an example of the protocol, we present an analysis of dopaminergic (DA) neurons in adult brains of WT (w1118) flies. The high efficacy of the dissection method makes it particularly useful for large-scale adult brain-based studies in Drosophila.

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

The adult Drosophila brain is a valuable system for studying neuronal circuitry, higher brain functions, and complex disorders. An efficient method to dissect whole brain tissue from the small fly head will facilitate brain-based studies. Here we describe a simple, one-step dissection protocol of adult brains with well-preserved morphology.

一个简单的一步解剖协议成人全安装准备<em&gt;果蝇</em&gt;脑

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the ...

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

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

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

时间依赖性增加对微电极阵列刺激神经元细胞培养物的网络响应

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

Application of Automated Image-guided Patch Clamp for the Study of Neurons in Brain Slices

Whole-cell patch clamp is the gold-standard method to measure the electrical properties of single cells. However, the in vitro patch clamp remains a challenging and low-throughput technique due to its complexity and high reliance on user operation and control. This manuscript demonstrates an image-guided automatic patch clamp system for in vitro whole-cell patch clamp experiments in acute brain slices. Our system implements a computer vision-based algorithm to detect fluorescently labeled cells and to target them for fully automatic patching using a micromanipulator and internal pipette pressure control. The entire process is highly automated, with minimal requirements for human intervention. Real-time experimental information, including electrical resistance and internal pipette pressure, are documented electronically for future analysis and for optimization to different cell types. Although our system is described in the context of acute brain slice recordings, it can also be applied to the automated image-guided patch clamp of dissociated neurons, organotypic slice cultures, and other non-neuronal cell types.

This protocol describes how to conduct automatic image-guided patch-clamp experiments using a system recently developed for standard in vitro electrophysiology equipment.

自动图像引导贴片夹在脑切片神经元研究中的应用

Whole-cell patch clamp is the gold-standard method to measure the electrical properties of single cells. However, the in vitro patch clamp remains a ...