A multiwell plate format microfluidic immobilization chip for high-content imaging of whole animals

用于整个动物高内涵成像的多孔板微流控固定芯片

• 批准号: 9901648
• 负责人: Evan Hegarty
• 金额: $ 5.5万
• 依托单位: VIVOVERSE, LLC
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2019-09-13
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9901648
• 关键词: Aging; Algorithmic Analysis; Algorithms; Animal Disease Models; Animal Model; Animals; Automation; Biological; Biological Assay; Caenorhabditis elegans; Cardiovascular system; Cells; Chemicals; Chronic Disease; Clinical; Collaborations; Custom; Development; Devices; Disease; Drug Costs; Drug Industry; Drug Screening; Failure; Functional disorder; Genes; Genetic; Glass; Glues; Health; Human; Human Biology; Image; Imaging Device; Immobilization; In Vitro; Injections; Lateral; Lead; Legal patent; Libraries; Liquid substance; Location; Longevity; Manuals; Methods; Microfluidic Microchips; Microfluidics; Modeling; Molds; Nerve Degeneration; Nervous system structure; Neuraxis; Neurons; Neurotoxins; Organ; Organoids; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phenotype; Plasma; Population; Positioning Attribute; Probability; Procedures; Process; Production; Proteins; Publishing; Reproducibility; Research; Resolution; Safety; Screening procedure; Secure; Signal Transduction; Small Business Innovation Research Grant; Speed; Standardization; System; Technology; Testing; Texas; Thinness; Time; Toxic effect; Translating; Translations; Universities; Variant; Whole Organism; age related; austin; automated image analysis; base; cost; data acquisition; density; design; drug discovery; experimental study; fluorescence imaging; genetic manipulation; high resolution imaging; high throughput screening; human disease; image processing; imaging system; improved; in vitro Assay; in vivo; industry partner; instrument; light weight; lithography; neurotoxicity; neurotoxicology; new technology; new therapeutic target; novel; off-patent; operation; polyglutamine; prevent; prototype; scale up; screening; seal; submicron; tool; user-friendly

Advancements in precise genetic manipulation have helped biologists to identify new drug-targets and in vivo disease mechanisms using small animal models, such as C. elegans. Human diseases pathophysiologies are reproduced in C. elegans expressing human disease genes inside the animal. Great opportunities are provided by the recent surge is genetic tools for animal models, but the lack of high-content screening (HCS) technologies precluded these models from screening for subtle phenotypes that better recapitulate the human disease situations. Development of novel technologies will enable the use of such models, at the same cost and speed of in vitro assays, to discover new drug targets and understand mode-of-action for new compounds in vivo. Dr. Ben-Yakar Lab at The University of Texas at Austin has developed a novel large-scale microfluidic chip that can image ~4,000 animals from 96 populations using a proprietary channel design. An efficient image acquisition and analysis algorithms can screen a whole chip within 16 min, a record speed that is 10,000× faster than manual imaging. To translate this lab prototype into marketplace, this SBIR Phase I application proposes to develop a beta model vivoChip-96x that will be with SBS format, compatible with automation, lighter weight, less expensive, and user-friendly to operate with the new top-gasket design. The new chip design will be bonded to a thin glass substrate at the bottom to enable improved imaging using high-resolution objectives. The proposed microfluidic chip will incorporate a machined top-plastic with custom-designed wells in a micro-titer format for easy integration to liquid handling systems for the high-throughput screen (HTS), and avoid substrate bending, and chip-handling errors. In Aim 1, we plan to develop a beta design of the vivoChip-96x device with the top- plastic piece, fabricate a thin PDMS layer using soft-lithography, and bond the interfaces using appropriate UV- cured glue and plasma treatment. The beta model will be operated with a new top-gasket with an improved sealing mechanism. C. elegans populations will be trapped inside the channels to characterize the variability in the trapping efficiency using four chips and following the standard operational procedures (SOPs). In Aim 2, we will develop an automated acquisition algorithm with BioTek to image C. elegans with high reslution objectives having a sub-cellular expression of fluorescent proteins and achieve an assay quality Z’~0.8. Achieving these milestones in Phase I, we will be able to reduce the current cost of the chip by 3 folds and standardize the vivoChip-96x for all commercially available HCS instruments. In Phase II, we will develop a fully automated vivoLoader to replace our current semi-automated worm handling procedures of liquid handling and an automated image-analysis platform (vivoAnalyzer) that will identify subtle fluorescent phenotype in low expressing C. elegans. Using our ongoing research collaboration, we plan to apply our screening technology to develop neurotoxicity and neurodegeneration assays to be able to screen novel compounds from large pharmaceutical companies. Support from industry partners will help us to translate the prototype into a product.

精确遗传操作的进步帮助生物学家识别新的药物靶点和体内 疾病机制，使用小动物模型，如C.优雅的人类疾病的病理生理学是 复制于C.在动物体内表达人类疾病基因的线虫。提供了巨大的机会 最近激增的是用于动物模型的遗传工具，但缺乏高含量筛选（HCS）技术 排除了这些模型筛选更好地概括人类疾病的微妙表型 situations.新技术的发展将使这些模型能够以同样的成本和速度使用 体外分析，以发现新的药物靶点，并了解新化合物在体内的作用模式。 德克萨斯大学奥斯汀分校的Ben-Yakar实验室开发了一种新型的大规模微流体芯片 它可以使用专有的通道设计对来自96个种群的约4,000只动物进行成像。高效的图像 采集和分析算法可以在16分钟内筛选出整个芯片，记录速度快10，000倍 而不是手动成像。为了将该实验室原型转化为市场，SBIR第一阶段应用程序建议 开发一个测试模型vivoChip-96 x，将与SBS格式，兼容自动化，重量更轻， 更便宜，并且使用新的顶部垫圈设计操作方便。新的芯片设计将结合 在底部的薄玻璃基板，以实现使用高分辨率物镜改善成像。拟议 微流控芯片将包括一个机加工的顶部塑料与定制设计的威尔斯在微量滴定格式， 易于集成到用于高通量筛选（HTS）的液体处理系统中，并避免基板弯曲， 和芯片处理错误。在目标1中，我们计划开发vivoChip-96 x设备的测试版设计， 塑料片，使用软光刻制作薄PDMS层，并使用适当的UV- 固化胶和等离子体处理。测试版车型将采用新的顶部垫圈， 密封机制C.线虫种群将被困在通道内，以表征 使用四个芯片并遵循标准操作程序（SOP）的捕获效率。在目标2中， 将与BioTek一起开发图像C的自动采集算法。具有高分辨率目标的elegans 具有荧光蛋白的亚细胞表达，并达到测定质量Z ′ ~ 0.8。 在第一阶段实现这些里程碑，我们将能够将芯片的当前成本降低3倍， 标准化vivoChip-96 x用于所有市售HCS仪器。在第二阶段，我们将全面发展 自动化vivoLoader取代我们目前的半自动化液体处理程序， 一个自动化的图像分析平台（vivoAnalyzer），将在低水平的细胞中识别微妙的荧光表型。 表达C.优美的利用我们正在进行的研究合作，我们计划将我们的筛选技术应用于 开发神经毒性和神经变性试验，以便能够从大规模筛选新化合物。 制药公司。来自行业合作伙伴的支持将帮助我们将原型转化为产品。