High-Throughput Cell Mechanical Property Testing for Label-Free Assaying

项目总结：我们引入了一种独特的基于微流控的方法来实现高通量 细胞的非破坏性分析，不需要特定的标记或试剂。基于 使用施加的光学力测量静态和动态单元机械特性， 我们将把这种技术(称为“光学拉伸”)应用于高速、高通量的 举止。到目前为止，光学拉伸仅用于小细胞数量；然而，高- 强度、微尺度激光光源及其在动态微流控中的集成 系统使我们提出的方法成为可能。在这方面，完全集成的基于光学的传感器和 将使用机械担架来识别并根据需要分离单个细胞。一次 这样的靶细胞然后可以在芯片上被输送到 或用于分配到标准生物实验室仪器中进行芯片外分析。 尽管有广泛的需求，我们提出的技术将进行测试和开发，使用 疟疾寄生虫以感染红细胞为靶细胞。这项工作将在 与NIAID疟疾和病媒研究实验室合作。我们的目标 包括：目标1：力学性能检测与解释。我们将采用光纤 集成在微流控系统中的用于无标记、非破坏性细胞的操作方法 机械性能测量。将为这两种情况开发建模方法 外力/变形实验数据的解释和设备设计。这里, 感染疟疾的红细胞将提供一个很好的模型靶点，因为细胞僵硬会发生变化 在寄生虫发育过程中戏剧性地。演示极大简化的设备设计和 相关联的易用性，我们将在NIH活跃的实验室中安装一台仪器。目标2：光学 用于细胞鉴定和分离的操作。我们将把光学方法整合到 用于单细胞检测和操作的微流控系统。在这里，两种芯片上的方法 细胞分离和芯片外分离将被开发和使用，以改善我们安装的NIH 原型。目标3：高吞吐量机械测试。为了实现高吞吐量， 将需要改进的微流控技术和更快的检测技术。在这个阶段，耦合 将探索流体动力和光学力的组合，以提高设备的性能。此外, 将使用时变的光学力来识别最佳信号响应和动态 物理属性。

Project Summary: We introduce a unique microfluidic-based approach for the high-throughput non-destructive assaying of cells without the need for specific labels or reagents. Based on measurement of both static and dynamic cell mechanical properties using applied optical forces, we will apply this technique (known as "optical stretching") in a high-speed high-throughput manner. To date, optical stretching has been used only on small cell numbers; however, high- intensity, microscale laser sources and the integration of these within dynamic microfluidic systems has enabled our proposed approach. In this, fully integrated optical-based sensors and mechanical stretchers will be used to identify and, upon demand, isolate single cells. Once identified, such targeted cells can then be transported on-chip to culture chambers within the device or for dispensing into standard bio-laboratory instrumentation for off-chip analysis. Though there is broad need, our proposed technology will be tested and developed using malaria parasite infected red blood cells as the target cell. This work will be done in collaboration with the Laboratory of Malaria and Vector Research at the NIAID. Our aims include: Aim 1: Mechanical Property Detection and Interpretation. We will employ optical manipulation methods integrated within microfluidic systems for label-free, non-destructive cell mechanical property measurement. Modeling approaches will be developed for both interpretation of applied force/deformation experimental data and for device design. Here, malaria-infected red blood cells will provide a good model target since cell stiffness changes dramatically during parasite development. Demonstrating greatly simplified device designs and associated ease-of-use, we will install an instrument in an active NIH laboratory. Aim 2: Optical Manipulation for Cell Identification and Isolation. We will integrate optical methods within microfluidic systems for single cell detection and manipulation. Here, methods for both on-chip cell isolation and off-chip isolation will be developed and used to improve our installed NIH protototype. Aim 3: High Throughput Mechanical Testing. To achieve high-throughputs, modified microfluidic and faster detection techniques will be required. In this phase, the coupling of hydrodynamic and optical forces will be explored to improve device performance. In addition, time-varying optical forces will be employed to identify optimal signal response and dynamic physical properties.