High throughput behavior monitoring and screening station for zebrafish larvae

描述（由申请人提供）：斑马鱼是一种广泛用于发育生物学的小型昼行硬骨鱼，作为一种廉价、快速繁殖的神经系统疾病研究动物模型，正受到越来越多的关注。成对的成虫通常在一夜之间产下50到200个卵。幼鱼表现出足够宽的行为谱，可用于大脑和行为研究的高通量工具所需的基于行为的遗传或药理学筛选。到目前为止，由于幼鱼非常小（约7毫米长）和透明，进展速度受到阻碍;目前可用的监测工具价格昂贵，精度有限，并且需要将个体鱼隔离在单独的单元中。基于使用消费级摄像机捕获图像的原始方法，我们设计了一个概念（昵称为“Zebratrack”），并实现了视频记录站的8罐原型（罐是培养皿形状的30 mL容器），以监测、筛选和分析成群的斑马鱼，而不需要隔离个体鱼。Zebratrack能够长时间（几天）连续记录多个视频流，并具有足够的分辨率来跟踪和分析个体鱼，从而可以检测和分析行为趋势以及罕见的瞬时行为。该系统非常易于使用;在其当前版本中，设置一个包含8个60个鱼缸的屏幕只需不到10分钟。一个真实的时间模式可以实时监控鱼的活动，一个分析模块在一个关系数据库中生成数据;这个数据可以使用标准的网络技术来访问，以检索数据，运行统计数据，并重放图像序列。拟议的努力旨在扩大Zebratrack的初始能力，提供增强的数据分析支持，并提供一个能够并行筛选2400条鱼的80个水箱系统。基于原型验证的概念，我们打算建立一个高带宽，功能强大，强大，易于使用，开放，可扩展，高度并行的筛选站，以支持高通量的斑马鱼行为筛选和监测。我们相信，由此产生的系统将是大脑和行为研究的一个出色的高通量工具，也是神经科学和行为科学界的一个宝贵的筛选平台。 公共卫生相关性：我们已经设计了一个原型的视频记录站，以监测，屏幕和分析离合器的年轻斑马鱼在组中，而不需要单独的鱼隔离。基于原型验证的概念，我们建议建立一个高带宽，功能强大，鲁棒，易于使用，开放，可扩展，高度并行的筛选站，以支持高通量的斑马鱼行为筛选和监测。我们相信，由此产生的系统将是大脑和行为研究的一个出色的高通量工具，以及神经科学和行为科学界的一个宝贵的遗传和药理学筛选平台。

DESCRIPTION (provided by applicant): Zebrafish, a small diurnal teleost fish extensively used in developmental biology, is receiving increasing interest as an inexpensive, fast-reproducing animal model for the study of neurological disorders. Paired adults typically produce clutches of 50 to 200 eggs overnight. Young fish exhibit a spectrum of behavior wide enough to be used for behavior-based genetic or pharmacological screens of the type needed by high throughput tools for brain and behavior research. To date, the pace of progress has been hindered the fact that young fish are very small (approximately 7mm in length) and transparent; currently available monitoring tools are expensive, have limited precision, and require isolating individual fish in separate cells. Based on an original approach to capturing images using consumer grade video cameras, we have designed a concept (nicknamed "Zebratrack") and implemented an 8-tank prototype (tanks are Petri dish - shaped 30mL containers) of a video-recording station to monitor, screen and analyze clutches of zebrafish in group without requiring the isolation of individual fish. Zebratrack is capable of continuously recording multiple video streams in parallel over extended periods of time (several days) with sufficient resolution to track and analyze individual fish, so that behavioral trends as well as rare transient behaviors can be detected and analyzed. The system is extremely easy to use; in its current version, setting up a screen of eight 60-fish tanks takes just under 10 minutes. A real time mode enables live monitoring of the fish activity, and an analysis module generates data in a relational database; this data can be accessed using standard web technologies to retrieve data, run statistics, and replay image sequences. The proposed effort aims at expanding the initial capabilities of Zebratrack, at offering enhanced data analysis support, and at delivering an 80-tank system capable of screening in the order of 2400 fish in parallel. Based on the concepts proven with the prototype, we intend to build a high-bandwidth, powerful, robust, easy-to- use, open, scalable, highly-parallel screening station to support high throughput behavior screening and monitoring of zebrafish. We believe the resulting system will be an outstanding high throughput tool for brain and behavior research, as well as an invaluable screening platform for the neuroscience and behavioral science communities at large. PUBLIC HEALTH RELEVANCE: We have designed a prototype of a video-recording station to monitor, screen and analyze clutches of young zebrafish in group without requiring the isolation of individual fish. Based on the concepts proven with the prototype, we propose to build a high- bandwidth, powerful, robust, easy-to-use, open, scalable, highly-parallel screening station to support high throughput behavior screening and monitoring of zebrafish. We believe the resulting system will be an outstanding high throughput tool for brain and behavior research, as well as an invaluable genetic and pharmacological screening platform for the neuroscience and behavioral science communities at large.