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

摘要 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 发现号。由于他们的全基因组测序，与人类的基因同源性，非常好的特征 线虫的解剖学和实验室培养的简便性是研究基础生物学的理想模式生物。 并对各种人类疾病的病理机制提供了重要的见解。许多研究成果 涉及线虫需要基于图像的分析和随后基于年龄等因素的分类 蠕虫或表型差异。尽管存在几种基于成像的分析平台，但它们或 缺乏高分辨率3D成像能力，价格高得令人望而却步，生物兼容性低，和/或 时间和劳动密集度太高，不适合高通量研究。这些缺点减损了 线虫研究的整体质量，特别是在药物发现方面，那里必须有数以万计的蠕虫 根据只有高分辨率成像才能检测到的细微差异进行快速分类。目标是 这个SBIR项目的目的是克服现有线虫成像和分选技术的限制，并 通过开发和商业化生物兼容蠕虫分析平台来解决市场上未得到满足的需求 能够使用声流控技术进行高分辨率、快速的线虫3D成像和分选(即融合 声学和微流体)技术。在我们第一阶段项目的工作中，我们成功地展示了 通过开发高性能的声流控芯片，验证了该技术的实用性和可行性。 吞吐量(155个蠕虫/分钟)、高存活率(~99.2%)以及高分辨率线虫3D成像和分选。在……里面 第二阶段，我们的商业化活动将集中于显著提高 基于声流控的3D成像和分类芯片，开发独立的、可进行Beta测试的原型， 并与最终用户一起验证和优化测试版原型。通过优化我们的声流控系统的设计 芯片，我们将提高吞吐量(&gt；1,000蠕虫/分钟)、旋转分辨率(&lt；1°)和分拣精度 (&gt；99%)，同时保持我们声流控技术的高生物兼容性(&gt；99%)。建议数 声流技术将成为唯一能够快速3D成像和 在单个设备上进行排序。通过实现对线虫的精确、快速的3D成像和自动分选， 以这种方式，我们的声流控技术可以满足包括制药在内的许多领域的关键需求 发展方面，到目前为止，由于技术限制，线虫在高科技领域的广泛使用受到阻碍。 吞吐量药物筛选。由于其优越的速度、精度、精确度和功能，我们相信 我们的声流控技术将成为C。 并将有助于简化工作流程、扩展功能并加快在多个领域的进展 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.