Novel System for Automation of Zebrafish Lipid Assays

描述（由申请人提供）：斑马鱼，Danio rerio，已被广泛接受为脊椎动物发育的有用模型。人们对开发斑马鱼作为一种模式生物用于高通量药物发现分析有浓厚的兴趣。到目前为止，真正的高通量筛选斑马鱼仅限于胚胎和非常早期的幼虫。这是因为研究人员最感兴趣的许多器官最好是从动物的侧面观察。一旦鱼鳔膨胀，幼虫通常直立漂浮，很难产生动物侧面的清晰图像。在多孔微孔板格式中尤其如此。解决这一重要问题的努力集中在使用共聚焦成像技术或强大的图像反卷积软件来组装动物的三维重建。这些努力并没有达到预期的空间分辨率，通常需要1到2分钟才能对单井进行成像。物理科学公司（PSI）发明了一种新技术，该技术将适用于已经在药物发现实验室使用的标准图像分析系统。PSI的技术将允许研究人员在微孔阵列中生成老斑马鱼幼虫的清晰侧面视图。该技术不需要共聚焦显微镜技术或复杂的图像反卷积算法，因此可以在几秒钟内生成高质量的图像，而其他技术每口井需要几分钟。在第一阶段，我们将使用Zygogen公司正在开发的Z-Lipotrack检测来证明我们技术的可行性。之所以选择脂道法，是因为它通常是在年龄较大的幼虫上进行的，并且需要收集胆囊和肠道的图像。在第二阶段，我们将进一步开发PSI的创新成像技术，并将该技术整合到一个完全集成的自动化斑马鱼幼虫发现系统中。PSI的技术将利用Z-Lipotrack检测技术实现生物活性化合物的高通量发现，从而降低胆固醇和脂质水平。这些药物满足了一项重要的卫生保健需求。

DESCRIPTION (provided by applicant): The zebrafish, Danio rerio, has become widely accepted as a useful model for vertebrate development. There is strong interest in developing zebrafish as a model organism for use in high throughput drug discovery assays. Thus far, true high throughput screening in zebrafish has been limited to embryos and very early larvae. This is because many of the organs that are most interesting to researchers are best viewed from the side of the animal. Once the swim bladder inflates and the larvae generally float upright it is difficult to generate a clear image of the side of the animal. This is especially true in multiwell microplate format. Efforts to resolve this important problem have centered on use of confocal imaging techniques or powerful image deconvolution software to assemble 3-D reconstructions of the animal. These efforts have not achieved the desired spatial resolution and typically require 1 to 2 minutes to image a single well. Physical Sciences Inc. (PSI) has invented a novel technology that will be adaptable to standard image analysis systems that are already in use in drug discovery laboratories. PSI's technology will allow researchers to generate clear side- views of older zebrafish larvae in microwell arrays. The technology does not require confocal microscopy techniques or sophisticated image deconvolution algorithms and will therefore generate quality images in seconds per well versus minutes per well for those other techniques. In Phase I we will use the Z-Lipotrack assay under development at Zygogen Corp to demonstrate the feasibility of our technology. The Lipotrack assay was chosen because it is typically performed on older larvae and requires collection of images of the gall bladder and intestine. In Phase II we will further develop PSI's innovative imaging technology and integrate that technology into a fully integrated, automated discovery system for zebrafish larvae. PSI's technology will enable high throughput discovery using the Z-Lipotrack assay of bioactive compounds that reduce cholesterol and lipid levels. Those drugs meet an important health care need.