Development of an acoustofluidic device for high-throughput 3D imaging and sorting of C. elegans

摘要 C.秀丽线虫是一种毫米长的透明蠕虫，广泛用于生物医学研究和药物 的发现由于它们的基因组完全测序，与人类的遗传同源性， 解剖结构和易于实验室培养，C.线虫是研究基础生物学的理想模式生物 这些研究为各种人类疾病的病理机制提供了关键的见解。许多研究工作 涉及C.秀丽线虫需要基于图像的分析和随后的排序的基础上的因素，如年龄的 蠕虫或表型差异。尽管存在几种基于成像的分析平台，但它们 缺乏高分辨率3D成像能力，过于昂贵，生物相容性低，和/或 太费时费力，不适合高通量研究。这些缺点有损于 C的整体质量。线虫的研究，特别是在药物发现，在成千上万的蠕虫必须 根据只有高分辨率成像才能检测到的微小差异进行快速分类。客观 这个SBIR项目的目的是克服现有C. elegans成像和分选技术， 通过开发和商业化生物相容性蠕虫分析平台，满足市场上未满足的需求 高分辨率、快速C.使用声流体的秀丽线虫3D成像和分选（即，的融合 声学和微流体）技术。在第一阶段的工作中，我们成功地证明了 该技术的实用性和可行性，通过开发能够高， 通量（155蠕虫/分钟）、高存活率（~99.2%）和高分辨率C. elegans 3D成像和分选。在 在第二阶段，我们的商业化活动将集中于显著提高 基于声流体的3D成像和分选芯片，开发独立的，beta测试就绪的原型， 并与最终用户一起验证和优化beta原型。通过优化我们的声流体的设计 芯片，我们将提高吞吐量（>1,000蠕虫/分钟），旋转分辨率（<1°），和分拣精度 （>99%），同时保留了我们声流技术的高生物相容性（>99%）。拟议 声流体技术将是唯一能够快速3D成像的自动蠕虫分析平台， 在单个设备上进行排序。通过精确、快速的3D成像和C. elegans在一个自动的 我们的声流体技术可以满足许多领域的关键需求，包括制药， 发展，其中技术限制迄今为止阻止了C.在高- 药物筛选。由于其上级速度、精度、准确性和功能，我们相信， 我们的声流技术将成为C. 优雅，将有助于简化工作流程，扩大能力，并加快在许多领域的进展， 生物医学研究

Abstract C. elegans are millimeter-long, transparent worms that are widely used in biomedical research and drug discovery. Due to their fully sequenced genome, genetic homology with humans, extremely well characterized anatomy, and ease of lab cultivation, C. elegans are an ideal model organism for studying fundamental biological processes and have provided key insights into the pathologies of various human diseases. Many research efforts involving C. elegans require image-based analysis and subsequent sorting based on factors such as the age of the worms or phenotypic differences. Although several platforms for imaging-based analysis exist, they either lack high-resolution 3D imaging capabilities, are prohibitively expensive, have low biocompatibility, and/or are far too time and labor intensive to be suitable for high-throughput studies. These drawbacks detract from the overall quality of C. elegans research, particularly in drug discovery, where tens of thousands of worms must be rapidly sorted based on minute differences that can only be detected with high-resolution imaging. The objective of this SBIR project is to overcome the limitations of existing C. elegans imaging and sorting technologies and address unmet needs in the market by developing and commercializing a biocompatible worm analysis platform capable of high-resolution, rapid C. elegans 3D imaging and sorting using acoustofluidic (i.e., the fusion of acoustics and microfluidics) technologies. During our work on the Phase I project, we successfully demonstrated the utility and feasibility of the proposed technology by developing acoustofluidic chips capable of high- throughput (155 worms/min), high-viability (~99.2%), and high-resolution C. elegans 3D imaging and sorting. In Phase II, our commercialization activities will focus on significantly improving the performance of the acoustofluidic-based 3D imaging and sorting chips, developing self-contained, beta-testing-ready prototypes, and validating and optimizing the beta prototypes with end users. By optimizing the design of our acoustofluidic chips, we will improve the throughput (>1,000 worms/min), rotational resolution (<1°), and sorting accuracy (>99%), while retaining the high biocompatibility (>99%) of our acoustofluidic technology. The proposed acoustofluidic technology will be the only automated worm analysis platform capable of rapid 3D imaging and sorting on a single device. By enabling precise, rapid 3D imaging and sorting of C. elegans in an automated manner, our acoustofluidic technology can address critical needs in many fields including pharmaceutical development, where technological limitations have thus far prevented the widespread use of C. elegans in high- throughput drug screening. Due to its superior speed, precision, accuracy, and functionalities, we believe that our acoustofluidic technology will become the platform of choice for the imaging, analysis, and sorting of C. elegans and will help to streamline workflows, expand capabilities, and accelerate progress in many areas of biomedical research.