Microcyclone arrays for high resolution bioaerosol fractionation and viable virus collection

项目摘要 我们的目标是开发一种多级微尺度旋风分离器技术， 生物气溶胶分离平台，使基础和应用病毒空气生物学的研究更加有效。的 微旋风分离器将被设计成分离从呼出气样品收集的选定的气溶胶尺寸级分， 使得能够以高尺寸分辨率评估这些样品中呼吸道病毒的分布。值得注意的是， 该技术的设计将克服现有空气生物学仪器的局限性， 动态尺寸范围、收集的级分的最大数量、分辨率、通量和生物效率。平台 将利用基于高分辨率立体光刻的3D打印技术来图案化大阵列， 在单片基底中的复杂微旋流器结构，具有串联放置的多个阵列，以允许选择 气溶胶尺寸范围从样品流中分离出来。该系统将进一步支持水凝胶的整合 在每个微旋流器阵列内设置一层，以捕获活病毒用于感染性研究。微旋流器阵列 将与已建立的呼出气收集系统相结合，并用于研究 流感病毒从受感染的受试者。为了支持这些目标，将设计单独的微旋流器元件， 制造、表征和优化用于分离至少五种不同的气溶胶尺寸级分， 10 µm，随后开发了集成数百个独立分离元件的完整微旋流器阵列 在单个设备中。一组级联阵列将组装到可重复使用的盒中，以收集所有目标 在一个单一的集成单元内的馏分，并产生的墨盒将与现有的系统接口， 呼出气体积采集（米尔顿实验室开发的G-II采样器）。综合文书将 用于采集至少15名活动性流感感染者的呼出气样本，以进行评估 病毒在所有粒级中的分布。研究结果预计将验证该技术作为一种强大的 一种提高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 一系列的空气传播疾病。成功地完成这些目标将使人们清楚地看到 从呼出气中收集和高分辨率分离生物气溶胶的微旋流器阵列技术 样品对于未来的步骤，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， 扩大阵列的规模，以支持更高流速的环境采样，并扩大该技术的应用 气溶胶化的细菌和真菌孢子的表征。

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.