Ramazani Roseanna B, Krishnan Harish R, Bergeson Susan E, Atkinson Nigel S
Section of Neurobiology and Institute for Neuroscience, University of Texas at Austin, 1 University Station, Austin, TX 78712, USA.
J Neurosci Methods. 2007 May 15;162(1-2):171-9. doi: 10.1016/j.jneumeth.2007.01.005. Epub 2007 Jan 16.
Currently, measuring ethanol behaviors in flies depends on expensive image analysis software or time intensive experimental observation. We have designed an automated system for the collection and analysis of locomotor behavior data, using the IEEE 1394 acquisition program dvgrab, the image toolkit ImageMagick and the programming language Perl. In the proposed method, flies are placed in a clear container and a computer-controlled camera takes pictures at regular intervals. Digital subtraction removes the background and non-moving flies, leaving white pixels where movement has occurred. These pixels are tallied, giving a value that corresponds to the number of animals that have moved between images. Perl scripts automate these processes, allowing compatibility with high-throughput genetic screens. Four experiments demonstrate the utility of this method, the first showing heat-induced locomotor changes, the second showing tolerance to ethanol in a climbing assay, the third showing tolerance to ethanol by scoring the recovery of individual flies, and the fourth showing a mouse's preference for a novel object. Our lab will use this method to conduct a genetic screen for ethanol-induced hyperactivity and sedation, however, it could also be used to analyze locomotor behavior of any organism.
目前，测量果蝇的乙醇行为依赖于昂贵的图像分析软件或耗时的实验观察。我们设计了一个用于收集和分析运动行为数据的自动化系统，使用IEEE 1394采集程序dvgrab、图像工具包ImageMagick和编程语言Perl。在所提出的方法中，将果蝇放置在一个透明容器中，一台计算机控制的相机定期拍照。数字减法去除背景和静止的果蝇，在发生运动的地方留下白色像素。对这些像素进行计数，得出一个与图像之间移动的动物数量相对应的值。Perl脚本使这些过程自动化，从而与高通量基因筛选兼容。四个实验证明了该方法的实用性，第一个实验展示了热诱导的运动变化，第二个实验在攀爬试验中展示了对乙醇的耐受性，第三个实验通过对单个果蝇的恢复情况评分展示了对乙醇的耐受性，第四个实验展示了小鼠对新物体的偏好。我们实验室将使用这种方法对乙醇诱导的多动和镇静进行基因筛选，然而，它也可用于分析任何生物体的运动行为。
