Tracking movement behavior of multiple worms on food.
Tracking movement behavior of multiple worms on food.
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DOI：
10.1101/pdb.prot067025
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发表时间：
2011-12-01
影响因子：
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通讯作者：
Schafer, William R
中科院分区：
文献类型：
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作者：
Yemini, Eviatar;Kerr, Rex A;Schafer, William R
遗传易驯化生物的神经生物学研究在很大程度上依赖于对行为表型的可靠分析。线虫线虫简单的身体结构使其特别适合使用自动显微镜和图像分析来定量描述行为模式。正向遗传筛选和药物库筛选需要高通量的表型分析，这项任务传统上与人工对数量变化的行为进行评分不兼容。现在，通过几个不同的跟踪软件包，高通量自动分析线虫的运动行为是可能的。这里介绍的多蠕虫跟踪器(MWT)是为高通量分析而设计的：它可以同时以每秒30帧的速度记录数十个蠕虫，一次持续数小时或数天。这是通过实时执行所有图像分析来实现的，仅将蜗杆质心、轴承和轮廓数据保存到磁盘。为了简化图像处理，该系统只关注已经移动的蠕虫，并检测并丢弃接触到的蠕虫，而不是试图通过计算将它们隔离。由于该软件是完全自动化的，一旦放置了蠕虫并启动了软件，协议就可以在无人值守的情况下运行。MWT不会保存图像以供以后分析，但可以使用附带的分析工具手动验证行为，该工具可以回放记录的身体姿势。该方案描述了使用MWT对食物进行基本的基本运动分析；类似的方案适用于相关的分析和类似的多种动物追踪器。该方案可以扩展到从TAP反应到趋化作用的各种检测。
Neurobiological research in genetically tractable organisms relies heavily on robust assays for behavioral phenotypes. The simple body plan of the nematode Caenorhabditis elegans makes it particularly amenable to the use of automated microscopy and image analysis to describe behavioral patterns quantitatively. Forward genetic screens and screens of drug libraries require high-throughput phenotyping, a task traditionally incompatible with manual scoring of quantitatively varying behaviors. High-throughput automated analysis of C. elegans movement behavior is now possible with several different tracking software packages. The Multiworm Tracker (MWT) described here is designed for high-throughput analysis: it can record dozens of worms simultaneously at 30 frames per second for hours or days at a time. This is accomplished by performing all image analysis in real time, saving only the worm centroid, bearing, and outline data to the disk. To simplify image processing, the system focuses only on worms that have moved, and detects and discards worms that are touching rather than trying to isolate them computationally. Because the software is entirely automated, protocols can run unattended once the worms have been placed and the software has been started. The MWT does not save images for later analysis, but behavior can be validated manually with a companion analysis tool that replays recorded body postures. This protocol describes a basic basal movement assay on food using the MWT; similar protocols apply to related assays and to similar multiple animal trackers. The protocol can be extended to a variety of assays ranging from tap response to chemotaxis.