Automated Screening System for Modifiers of Cardiac Output in Zebrafish

描述（由申请人提供）：斑马鱼，Danio rerio，已被广泛接受为人类疾病的模型。已经进行了小分子筛选，以确定正常心血管过程的调节剂和心血管疾病的调节剂。正在开发的高通量系统旨在测量心率，但不解决心血管功能的其他关键参数，如血流速度和分布。我们建议开发一种高通量系统，它可以同时：1)测量心率；2)生成主要血管的血流图；3)测量血管中的血流速度。基于显微镜的系统将利用多普勒光谱域光学相干断层扫描，在几秒钟内生成斑马鱼胚胎和幼虫的三维结构图像，并测量血液流量。在第一阶段，我们将开发一个面包板系统，该系统将能够从动物的背部和腹部视图收集图像。大脑和尾巴的血管图像将被收集。在面包板系统中，动物将被定位，扫描角度将被手动调整。在第二阶段，将开发一个完整的自动化筛选系统，包括：1)软件算法，快速定位动物进行成像，并沿其轴向长度调整扫描角度；2)自动图像采集；3)自动图像处理。二期试验将包括一个先导化合物筛选，以确定心输出量的调节剂。

DESCRIPTION (provided by applicant): The zebrafish, Danio rerio, has become widely accepted as a model for human disease. Small molecule screens have been performed to identify both modifiers of normal cardiovascular processes and modifiers of cardiovascular disease. High-throughput systems under development aim to measure heart rate but do not address other critical parameters of cardiovascular function such as blood velocity and distribution. We propose to develop a high-throughput system that would simultaneously: 1) measure heart rate; 2) generate a flow map of the major blood vessels; and 3) measure blood velocity in those vessels. The microscope-based system will utilize spectral domain optical coherence tomography with Doppler to generate, within a matter of seconds, a three dimensional structural image of zebrafish embryos and larvae and measure blood flow. In Phase I we will develop a breadboard system that will enable collection of images from both the dorsal and ventral views of the animals. Images of the vasculature both of the brain and of the tail will be collected. In the breadboard system the animals will be located and the scan angle will be adjusted manually. In Phase II a complete, automated screening system will be developed that includes: 1) software algorithms to rapidly locate the animals for imaging and adjust the scan angle along their axial length; 2) automated image collection; and 3) automated image processing. Phase II testing will include a pilot compound screen to identify modifiers of cardiac output. This research will develop technologies that will enable the use of zebrafish models of human disease in high-throughput screens for active pharmaceutical compounds. In particular, such screens are expected to accelerate discovery of new pharmaceuticals to treat cardiovascular diseases.