Microcyclone arrays for high resolution bioaerosol fractionation and viable virus collection

项目总结 我们的目标是开发一种多级微尺度旋风技术，作为一种高效和可扩展的 生物气溶胶分级平台，使基础和应用病毒空气生物学的研究更有效。这个 微型旋风分离器将被设计成分离从呼气样本中收集的选定的气溶胶尺寸组分， 使呼吸道病毒在这些样本中的分布能够以高尺寸分辨率进行评估。值得注意的是， 这项技术的设计将克服现有有氧生物仪器的局限性，通过增强 动态大小范围、收集的最大分级数、分辨率、吞吐量和生物效率。站台 将利用基于高分辨率立体平版印刷的3D打印技术来为大型阵列 单块衬底中的复杂微旋风结构，多个阵列串联放置，以允许选择 气雾剂的大小范围要与样品流隔离。该系统将进一步支持水凝胶的集成 每个微旋风阵列内的一层，以允许捕获用于传染性研究的活病毒。微旋风分离器阵列 将与已建立的呼气收集系统相结合，并用于研究 来自受感染对象的流感病毒。为了支持这些目标，将设计单独的微旋风元件， 制造、表征和优化分离至少五种不同的气溶胶粒度组分，范围从200 nm到 10微米，随后开发了集成数百个独立分离元件的全微旋风阵列 在一台设备中。一组级联阵列将组装成可重复使用的盒式磁带，以实现对所有目标的收集 单个集成单元内的馏分，所产生的盒将与现有系统对接，以实现高- 呼气量采样器(弥尔顿实验室开发的G-II采样器)。集成的仪器将 用于收集至少15名活动性流感感染者的呼出呼气样本进行评估 病毒分布在所有大小的组分上。预计研究结果将验证该技术是一种强大的 增强我们对空气生物学的理解并改进建模、风险分析和缓解策略的工具 一系列通过空气传播的疾病。成功完成这些目标将使我们清楚地看到 用于收集和高分辨率分离呼气生物气溶胶的微旋风阵列技术 样本。对于未来的步骤，我们设想优化该技术的生物效率，以支持收集的病毒的培养， 扩展阵列以支持更高流速下的环境采样，并扩展该技术的应用 气雾化细菌和真菌孢子的特性。

PROJECT SUMMARY Our objective is to develop a multi-stage microscale cyclone technology that will serve as an efficient and scalable platform for bioaerosol fractionation, enabling more effective studies of fundamental and applied viral aerobiology. The microcyclones will be designed to isolate selected aerosol size fractions collected from exhaled breath samples, enabling the distribution of respiratory virus within these samples to be evaluated with high size resolution. Significantly, the technology will be designed to overcome the limitations of existing aerobiological instruments by enhancing the dynamic size range, maximum number of collected fractions, resolution, throughput, and bioefficiency. The platform will take advantage of a high resolution stereolithography-based 3d printing technique to pattern large arrays of complex microcyclone structures in a monolithic substrate, with multiple arrays placed in series to allow selected aerosol size ranges to be isolated from the sample flow. The system will further support the integration of a hydrogel layer within each microcyclone array to allow the capture of live virus for infectivity studies. The microcyclone arrays will be combined with an established exhaled breath collection system and employed to study the distribution of influenza virus from infected subjects. To support these goals, individual microcyclone elements will be designed, fabricated, characterized, and optimized for isolating at least five distinct aerosol size fractions ranging from 200 nm to 10 µm, followed by the development of full microcyclone arrays integrating hundreds of individual separation elements in a single device. A set of cascaded arrays will be assembled into a reusable cartridge to enable collection of all target fractions within a single integrated unit, and the resulting cartridge will be interfaced with the existing system for high- volume exhaled breath collection (the G-II sampler developed in the Milton laboratory). The integrated instrument will be used to collect exhaled breath samples from at least 15 individuals with active influenza infection for the evaluation of virus distribution across all size fractions. The study results are expected to validate the technology as a powerful tool for enhancing our understanding of aerobiology and improving modeling, risk analysis, and mitigation strategies for a wide range of airborne diseases. Successful completion of these aims will offer a clear view of the potential of the microcyclone array technology for collection and high resolution fractionation of bioaerosols from exhaled breath samples. For future steps we envision optimizing bioefficiency of the technology to support culture of collected virus, scaling the arrays to support environmental sampling at higher flow rates, and extending application of the technology to the characterization of aerosolized bacteria and fungal spores.