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

用于无标记测定的高通量细胞机械特性测试

基本信息

批准号：
8305759
负责人：
CHARLES Dionisio EGGLETON
金额：
$ 31.73万
依托单位：
COLORADO SCHOOL OF MINES
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-15 至 2014-07-31
项目状态：
已结题

项目摘要

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.

项目概述：我们介绍了一种独特的基于微流体的高通量方法，不需要特定标记或试剂的细胞的非破坏性测定。基于使用施加的光学力测量静态和动态细胞机械性能，我们将把这种技术（称为“光学拉伸”）应用于高速高通量方式迄今为止，光学拉伸仅用于小细胞数;然而，高- 强度、微尺度激光源以及这些在动态微流体中的集成系统实现了我们提出的方法。在这方面，完全集成的光学传感器和将使用机械拉伸器来鉴定并根据需要分离单细胞。一旦鉴定后，然后可以将这样的靶细胞在芯片上运输到培养室内的培养室。设备或用于分配到标准生物实验室仪器中以进行芯片外分析。虽然有广泛的需求，我们提出的技术将进行测试和开发，疟疾寄生虫感染的红细胞作为靶细胞。这项工作将在与NIAID的疟疾和病媒研究实验室合作。我们的目标包括：目标1：机械性能检测和解释。我们将采用光学集成在微流控系统内的无标记、非破坏性细胞的操作方法机械性能测试将为这两个方面制定建模方法解释所施加的力/变形实验数据和用于装置设计。在这里，感染疟疾的红细胞将提供一个很好的模型目标，因为细胞硬度的变化在寄生虫发育过程中发生了巨大变化。展示了大大简化的设备设计，相关的易用性，我们将在一个活跃的NIH实验室安装一台仪器。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体系统。在这里，用于片上细胞隔离和芯片外隔离将被开发和用于改善我们安装的NIH 原型目标3：高吞吐量机械测试。为了实现高吞吐量，将需要改进的微流体和更快的检测技术。在这个阶段，耦合将探索流体动力学和光学力的作用，以提高器件性能。此外，本发明还提供了一种方法，时变光学力将用于识别最佳信号响应和动态物理性质