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

项目总结 此R01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化验，从而消除误差 与手动移液相关，并显著提高了重复性。然而，大多数自动化的 液体处理技术受到一个基本限制：它们依赖于与固体的物理接触 结构，以便操纵液体试剂。因此，微量的试剂不可避免地吸附在触点上。 并可能在以后溶解到另一个液体样本中。因此，由于以下原因造成交叉污染的风险 这种不受欢迎的“表面污垢”将输送表面限制为单一类型的工作液体和试剂。 组合。最近，我们发明了数字声流控(DAF)，一种基于声学的，可编程的，接触式的 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：研究方法。在这个R01项目中，我们将开发和验证具有以下功能的DAF流体处理系统 特点：(1)可重写性、可编程性和执行复杂、级联反应的能力：我们将 演示DAF沿规定的任意路线运输和混合“射流位”(即液滴)的能力 没有交叉污染，导致允许的试剂组合数量增加104倍 单一设备上的输入(与传统平台相比)；(2)生物兼容性：而不是 直接受到强大的声压或高电场的影响，液滴在 非接触式的，温柔的举止。我们的初步结果表明，DAF平台对 细胞的活力；(3)多功能性：DAF不限于具有特定声学、电学、流体力学或 磁性。这种多功能性使DAF适合处理广泛的液体，甚至是具有挑战性的 诸如低极性流体(例如，有机溶剂)、粘性或粘性样品(例如，血液和痰)的样品， 和固体(例如，粪便样本和模式生物)；(4)小型化和方便集成：我们的 与现有的DAF平台相比，DAF平台提供了前所未有的小型化和成本效益 机器人液体处理系统。此外，它还设计成与各种多井板集成在一起， 使其能够无缝地集成到现有的生物医学研究实验室中。使用前面提到的 优势，建议的DAF技术有可能超过当前的行业标准，解决 ，并为开发健壮、可重写、高性能的 吞吐量和数字可编程射流处理器。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DAF的深远潜力，以改进从临床到临床等领域的研究 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.