Development of a digital acoustofluidic system for automating liquid handling in biomedical research

项目摘要 此R 01应用程序响应NIH倡议PAR-19-253“重点技术研究和 发展”。自动化液体处理技术在生物医学研究的许多领域都很有价值。为 例如，机器人移液系统已被广泛用于自动化测定，从而消除错误 与手动移液相关，并显著提高了再现性。然而，大多数自动化 液体处理技术受到一个基本的限制：它们依赖于与固体的物理接触 结构，以操纵液体试剂。因此，痕量的试剂不可避免地吸附到接触件上 表面，并且可能稍后溶解到另一种液体样品中。因此，交叉污染的风险，由于 这种不希望的“表面结垢”将输送表面限制为单一类型的工作液体加上试剂 组合.最近，我们发明了数字声流控技术（Digital acoustofluidics，简称ACF），这是一种基于声学的、可编程的接触式传感器， 免费的液体处理技术，克服了与现有液体处理相关的关键障碍 方法.在这个R 01项目中，我们将开发和验证一个具有以下功能的可再生流体处理系统： 特点：（1）可重写性，可编程性，以及执行复杂级联反应的能力：我们将 证明了微流控器传输和混合“流体比特”的能力（即，液滴）沿着规定的任意路线 无交叉污染，导致允许的试剂组合数量增加104倍 在单个设备上输入（与传统平台相比）;（2）生物相容性： 直接受到强声压或高电场的作用，液滴被操纵在 非接触式的温和方式我们的初步结果表明，该平台没有显着的影响， （3）多功能性：生物传感器不限于具有特定声学、电学、流体动力学或生物学特性的流体。 磁性这种多功能性使其适合处理各种液体，即使是具有挑战性的 样品如低极性流体（例如，有机溶剂），粘性或粘稠的样品（例如，血和痰）， 和固体（例如，粪便样品和模式生物）;（4）小型化和方便的集成：我们的 与现有技术相比， 机器人液体处理系统此外，它被设计为与各种多孔板集成， 使其能够无缝集成到现有的生物医学研究实验室中。与上述 优势，拟议的技术有可能超过目前的行业标准，解决 在外地未得到满足的需求，并提供一个引人注目的平台，发展一个强大的，可持续的，高 吞吐量和数字可编程流体处理器。我们将在两个已建立的 生物医学应用：蛋白质晶体化学和高通量药物筛选。在这方面，我们的目标是 展示了生物医学的深远潜力，以改善从临床到临床的研究领域， 从化学到基础生物学

PROJECT SUMMARY This R01 application is responsive to the NIH initiative PAR-19-253 “Focused Technology Research and Development”. Automated liquid handling technologies are valuable in many areas of biomedical research. For example, robotic pipetting systems have been extensively utilized to automate assays, thereby eliminating errors associated with manual pipetting and significantly improving reproducibility. However, the majority of automated liquid handling technologies suffer from a fundamental constraint: they rely on physical contact with a solid structure in order to manipulate liquid reagents. Therefore, traces of a reagent inevitably adsorb onto the contact surface and can possibly later dissolve into another liquid sample. Thus, the risk of cross-contamination due to this undesirable “fouling of the surface” limits the transport surfaces to a single type of working liquid plus reagent combination. Recently, we invented digital acoustofluidics (DAF), an acoustic-based, programmable, contact- free, liquid handling technology, which overcomes the key obstacles associated with the existing liquid handling methods. In this R01 project, we will develop and validate a DAF fluid processing system with the following features: (1) Rewritability, programmability, and ability to perform complex, cascade reactions: We will demonstrate the ability of DAF to transport and mix ‘fluidic bits’ (i.e., droplets) along prescribed, arbitrary routes without cross-contamination, leading to a 104-fold increase in the number of allowable combinations of reagent inputs on a single device (as compared with conventional platforms); (2) Biocompatibility: Instead of being directly subjected to strong acoustic pressure or high electric fields, the droplets are manipulated in a contactless, gentle manner. Our preliminary results show that the DAF platform has no significant effect on the viability of cells; (3) Versatility: DAF is not restricted to fluids with specific acoustic, electrical, hydrodynamic, or magnetic properties. This versatility makes DAF suitable for handling a wide range of liquids, even for challenging samples such as low-polarity fluids (e.g., organic solvents), sticky or viscous samples (e.g., blood and sputum), and solids (e.g., fecal samples and model organisms); (4) Miniaturization and convenient integration: Our DAF platform provides an unprecedented level of miniaturization and cost-effectiveness compared with existing robotic liquid handling systems. In addition, it is designed to be integrated with a variety of multi-well plates, enabling it to be seamlessly integrated into existing biomedical research laboratories. With the aforementioned advantages, the proposed DAF technology has the potential to exceed current industry standards, address unmet needs in the field, and provide a compelling platform for the development of a robust, rewritable, high- throughput, and digitally-programmable fluidic processor. We will validate its performance across two established biomedical applications: protein crystal chemistry, and high-throughput drug screening. In this regard, we aim to demonstrate the far-reaching potential of DAF to enable improved research in areas ranging from clinical chemistry to fundamental biology.