This work was supported by NIH P20GM103643 to I. Meng.

1.准备幼虫<ol><li>为了分析运动或立场偏好所需的幼虫，在标准条件下生长的幼虫到所需的年龄标准使用飞食品11至10测定。 </li><li>请通过拉伸丝印级尼龙网在漏斗网状过滤器。固定网在漏斗颈部橡皮筋。放置在一个烧杯中的漏斗。 </li><li>为了收集幼虫进行分析，舀一铲满包含从培养瓶觅食幼虫的食物，并用RT自来水冲洗过的筛网过滤，收集直接个人幼虫网用画笔10。 </li></ol> 2.准备测定管<ol><li>要准备堵塞试管，精心熬了4％琼脂凝胶倒入培养皿中，以1.5厘米的深度。这将提供一个插头用于管的任一侧时动物被封闭。在4％的溶液是足够致密以防止由对幼虫enetrating插头。 </li><li>确保管是干净，清晰之前，插入幼虫。如果不是，这可能会阻止从被记录的动作。 </li><li>以保证幼虫有足够的水分运动，制备含有少量的热水挤压瓶。倒置的酒瓶，仔细定位走向水槽出口和压榨轻轻地从摄像管排出的水。 </li><li>将瓶子口进入测试管。挤压瓶子提供水汽进入测试管，直到薄层冷凝出现在管的壁。视频1示出了幼虫移动在管与水分的适当的量。 </li><li>以密封幼虫在管，取出从培养皿1.5厘米厚的琼脂凝胶，并放置在一网状表面，以允许气流琼脂下方以使得琼脂塞可插入到管。按该测定管的一端插入的凝胶两次，以插入两个插头凝胶成一端。 </li><li>随着paintb匆忙，将一个幼虫进入管约1.5英寸深，按最近的幼虫年底到琼脂凝胶密封管。所产生的压力将迫使出琼脂凝胶的第二插头从相对端和在管内密封该动物。 </li><li>地方管在MB5多波束果蝇活动监视器（DAM）装置，并调整管&#39;的位置，以使动物不能超越传感器的范围内。 </li><li>为了确保测定管在运输过程中不滑出设备的红外线读取帧时，通过紧固油灰的环每个管在此其接触记录装置周围将灯管保持到位。 </li></ol> 3.测量活动<ol><li>在培养箱中记录活动，以防止可能由于阴影，荧光灯或温度变化在实验室发生的不准确的读数。参见图5为系统的一个建议的布置。 </li><li>设置一个孵化至20℃（ 见图1）。为了避免虚假记载，由于从白炽灯光源的干扰录制过程中，关掉日光灯灯光孵化器，并使用单独的LED光源，同时记录在孵化器。没有任何动物进行试验，以确保光线条件并不异常触发的传感器。应该有这个测试后无记录的移动数据。 </li><li>让幼虫以适应到20℃培养箱设置开始试验前5分钟。 </li><li>要设置DAM系统以所需记录间隔（ 如 1分钟），确保大坝系统录音软件下载到电脑主机12和前打开连接到录音室到PSIU接口DAM系统文件。 </li><li>设置所需的记录频率数据将被保存，选择首选项，然后单击上方或读取间隔选项下方，选择不同的时间框架中的数据将被存储。例如，选择阅读的间隔时间从1秒到1小时。 </li><li>要选择不同的参数记录数据，选择首选项，然后选择相应的框中下输出数据类型（计数，移动，位置和纠缠，见表1）。每个参数提供在管内的幼虫活性的独特的分析。 </li><li>要开始录制，六硼化钙用电话线将显示器连接到电源接口单元（PSIU）。该PSIU连接到电源插座。绿灯会显示相应的连接。 </li><li>通过通用串行总线（USB）电缆到Macintosh或Windows PC进行数据记录连接PSIU。在电脑上打开DAM SystemMB1v6x程序自动开始记录数据。 </li><li>后所需采集时间完成后，选择退出，那么现在退出DAM活动屏幕上;数据将在DAM系统数据自动保存。借此数据文件（ 例如 ，监控1）并拖动<html>到一个单独的文件夹。这将从坝软件存储原始数据，使他们可以在以后进行处理。 </li></ol> 4.准备用于数据处理（DAM FileScan） <ol><li>为了处理原始数据转换成可以理解的格式打开DAM FileScan应用程序并选择选择输入数据的文件夹。然后选择数据文件夹和所需文件，并选择扫描选项。最后选择的bin长到所需的读取周期。这将原始数据组织成由操作者设定的参数，程序将报告收集在该特定范围内的数据。 </li><li>选择输出的数据类型来分析所需要的运动设定（监控计数，移动，住）。下的额外读数菜单选择总和成箱（这是如果仓长度大于所选择的系统的读取间隔）。 </li><li>适当命名的文件并保存。该文件将被存储在相同的位置，从步骤3.9的文件夹。 </li></ol>_title“&gt; 5。访问数据的分析<ol><li>为了收集平均移动，过程从第4步生成为了生成一个电子表格程序表中的原始数据，打开以前保存的文件（步骤4.3），并显示文本导入向导窗口时，选择完成打开在电子表格格式的数据文件。 注：根据分析的数据类型，电子表格将被读取不同。通常情况下测量，研究的时间段将数字记录监视器移动或计数，而每一个人的阅读管会收到一个指定的字母。当在电子表格中查看数据，列KZ表示用于分别测定管1-16的槽。该行表示所收集的数据点。 </li><li>例如，如果收集在1分钟的间隔20分钟的移动，平均的20行来计算移动/分钟等测量的平均数目。 </li></ol>

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The authors declare that they have no competing financial interests.

果蝇幼虫的活性是由多种因素，包括基因型8,13年龄和环境温度2的影响。虽然能够非常详细的分析，强大的录像制品的方法已经开发了那些谁学习运动5，这种详细程度可能是多余的为那些谁希望确定活动的基本参数。这里介绍的方法采用的设备是许多实验室使用，操作方便，产生高重现性的结果，即使是对于那些主要的研究重点是不运动管理。的例子的结果表明，...

The use of Drosophila as a genetic tool has transformed scientific knowledge of biological systems. Drosophila larvae have been used in a variety of studies including nociception1, development2 and as a model for the study of human disease genes3. Drosophila activity encompasses a range of behaviors that vary under different conditions including temperatures2, exposure to drugs4 and amongst different genotypes. Yet, despite the significant use of the larva as a model organism, a simple, standardized method to analyze larval activity has not been available. Presently, many larval locomotion studies employ sophisticated video analysis software5. While powerful, the complexity of such automated tools may discourage labs that are not already equipped to study locomotion from including analysis of informative activity parameters in their studies. In other current non-automated methods, such as the grid crawling analysis, motion is scored by a human observer, which introduces the possibility of subjectivity and limits throughput to one animal at a time6-7. A similar study used a 5-lane crawling assay, which measured the time it took larvae to travel a certain linear distance8. In such non-automated assays, displacement is measured but this does not account for non-linear travel between the start and end points. As discussed below, the method described here accounts for more of the actual larval movement, is objective, easy to operate, and offers robust throughput.
To easily study larval activity behavior without the compromise of accuracy, efficiency, or cost, this method employs the TriKinetics Drosophila Activity Monitor (DAM), a device often used to study adult activity. Using one device to study both adult and larval activity is cost-effective, and allows direct comparison of motion by animals at these two life stages. The system, featuring the manufacturer&#8217;s highest level of resolution, makes use of 17 infrared detection beams per assay tube, which record larval activity when sensor beams are broken within the 16 individual tubes. The system then automatically saves recorded information to a computer, making it available for manipulation with basic graphing software. The data obtained represents the beams that were broken by individual larvae (which can be converted into a rate), movement when a larva stays within a detector beam and the position of the animals within the assay chambers during a recording period (allowing one to calculate position preference). The system is efficient and relatively simple to operate, and brings highly reproducible basic activity analysis within the reach of any laboratory studying Drosophila larvae.
To demonstrate the power of this assay, data are presented that show its use to verify differences in activity resulting from varying ambient temperatures, as well as through the comparison of a mutant previously described as hypoactive (iav1)9 with a widely analyzed control (w1118).

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Drosophila Activity Monitor, Multibeam, 16 tubes, including wires</td>
			<td>TriKinetics Inc.&#160;</td>
			<td>MB5</td>
			<td></td>
		</tr>
		<tr>
			<td>Power Supply Interface for Activity Monitor&#160;</td>
			<td>TriKinetics Inc.&#160;</td>
			<td>PSIU24</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass 80 x 5 mm tubes for Activity Monitor (100)</td>
			<td>TriKinetics Inc.&#160;</td>
			<td>PGT 5x80</td>
			<td></td>
		</tr>
		<tr>
			<td>DAMsystemMB1v6x Data Acquisitions Software for Macintonsh OSX (Intel)</td>
			<td><a href="http://www.trikinetics.com/">www.trikinetics.com</a></td>
			<td></td>
			<td>free download</td>
		</tr>
		<tr>
			<td>DAMFileScan 108x software for Macintosh</td>
			<td><a href="http://www.trikinetics.com/">www.trikinetics.com</a></td>
			<td></td>
			<td>free download</td>
		</tr>
		<tr>
			<td>USB software (PSIUdrivers.zip).</td>
			<td><a href="http://www.trikinetics.com/">www.trikinetics.com</a></td>
			<td></td>
			<td>free download</td>
		</tr>
		<tr>
			<td>DAMSystem Notes 308</td>
			<td><a href="http://www.trikinetics.com/">www.trikinetics.com</a></td>
			<td></td>
			<td>free download</td>
		</tr>
		<tr>
			<td>Zeiss Stemi 2000C- Stereo Microscope</td>
			<td>Spectra Services</td>
			<td>SP-STEMI2000C-BS</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbon Dioxide</td>
			<td>Maine Oxy</td>
			<td></td>
			<td>anaesthesia</td>
		</tr>
		<tr>
			<td>Fly Pad</td>
			<td>Genesee</td>
			<td>59-114</td>
			<td>surface for sorting anaesthetized flies</td>
		</tr>
		<tr>
			<td>Small paint brush&#160;</td>
			<td>Winsor &#38; Newton</td>
			<td>#2 ROUND</td>
			<td>or similar, used for sorting anaesthetized flies</td>
		</tr>
		<tr>
			<td>Silk Screen Printing Mesh (160)</td>
			<td>msj-gallery.com</td>
			<td>SM160W63-3YD</td>
			<td>pore sized used in this protocol was ~ 0.1 mm</td>
		</tr>
		<tr>
			<td>Tegosept</td>
			<td>Genesee</td>
			<td>20-258</td>
			<td>preservative</td>
		</tr>
		<tr>
			<td>Ethanol (190proof)</td>
			<td>Pharmco</td>
			<td>111000190</td>
			<td>used to dissolve Tegosept</td>
		</tr>
		<tr>
			<td>6 oz Square Bottom Bottle (PP)</td>
			<td>Genesee</td>
			<td>32-130</td>
			<td></td>
		</tr>
		<tr>
			<td>&#34;Flugs&#34; for Plastic Fly bottles</td>
			<td>Genesee</td>
			<td>49-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Drosophila Vials, Wide (PS)</td>
			<td>Genesee</td>
			<td>32-117</td>
			<td></td>
		</tr>
		<tr>
			<td>Flugs for wide plastic vials</td>
			<td>Genesee</td>
			<td>49-101</td>
			<td></td>
		</tr>
		<tr>
			<td>Yellow Degerminated Corn Meal</td>
			<td>Gold Medal</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Drosophila agar</td>
			<td>LabScientific</td>
			<td>FLY 8020</td>
			<td></td>
		</tr>
		<tr>
			<td>Baker&#39;s Yeast - Red Star</td>
			<td>King Arthur Flour</td>
			<td>1270</td>
			<td></td>
		</tr>
		<tr>
			<td>Granulated Sugar - Extra Fine</td>
			<td>Domino</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

measurement of larval activity in the drosophila activity monitor

Drosophila larvae are used in many behavioral studies, yet a simple device for measuring basic parameters of larval activity has not been available. This protocol repurposes an instrument often used to measure adult activity, the TriKinetics Drosophila activity monitor (MB5 Multi-Beam Activity Monitor) to study larval activity. The instrument can monitor the movements of animals in 16 individual 8 cm glass assay tubes, using 17 infrared detection beams per tube. Logging software automatically saves data to a computer, recording parameters such as number of moves, times sensors were triggered, and animals&#8217; positions within the tubes. The data can then be analyzed to represent overall locomotion and/or position preference as well as other measurements. All data are easily accessible and compatible with basic graphing and data manipulation software. This protocol will discuss how to use the apparatus, how to operate the software and how to run a larval activity assay from start to finish.

图1示出了从控制第三龄幼虫的温度下反应的研究中，瓦特1118的结果，使用的监控装置，以检测在七个不同的温度下在幼虫运动的差异。幼虫洗涤并如上所述放入DAM活动装置，并放置在培养箱设置为所需的温度。该装置然后使其适应环境5分钟记录开始之前。每个幼虫单独为运动在20分钟内和每分钟移动平均数分析，计算为每一个动物，平均为每组32只动物。数据进行分析，并?...

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Measurement of Larval Activity in the Drosophila Activity Monitor

This report describes a method for measuring Drosophila larval activity using the TriKinetics Drosophila Activity Monitor. The device employs infrared beams to detect movements of up to 16 individual animals. Data can be analyzed to represent motion parameters including rates and the positions of the animals within the assay chambers.

幼虫活动的测量<em&gt;果蝇</em&gt;活动监视器

Drosophila larvae are used in many behavioral studies, yet a simple device for measuring basic parameters of larval activity has not been available. This ...

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Measurement of Larval Activity in the Drosophila Activity Monitor

9.2K 次观看.  University of New England. This report describes a method for measuring Drosophila larval activity using the TriKinetics Drosophila Activity Monitor. The device employs infrared beams to detect movements of up to 16 individual animals. Data can be analyzed to represent motion parameters including rates and the positions of the animals within the assay chambers.