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.

项目摘要：我们引入了一种独特的基于微流体的方法无需特定标签或试剂的细胞无损分析。基于使用施加的光学力测量静态和动态细胞机械性能，我们将在高速通量中应用此技术（称为“光伸展”）方式。迄今为止，光伸展仅用于小单元格数。但是，高 - 强度，显微镜激光源和这些集成在动态微流体中系统已使我们提出的方法。在此，完全集成的基于光学的传感器和机械担架将用于识别和根据需要隔离单细胞。一次鉴定出，可以将这种靶向细胞在芯片上运输到培养室内设备或分配到标准生物实验室仪器中进行片外分析。尽管有广泛的需求，但我们提出的技术将经过测试和开发疟疾寄生虫感染了红细胞作为靶细胞。这项工作将在与NIAID的疟疾实验室和媒介研究实验室合作。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体系统中的操纵方法，用于无标签，无损细胞机械性能测量。两者都将开发建模方法对应用力/变形实验数据和设备设计的解释。这里，疟疾感染的红细胞将提供一个良好的模型靶标，因为细胞刚度会发生变化在寄生虫发展期间急剧。展示了大量简化的设备设计和相关的易用性，我们将在活跃的NIH实验室中安装一种乐器。目标2：光学操纵细胞识别和分离。我们将在其中集成光学方法用于单细胞检测和操纵的微流体系统。在这里，两种芯片的方法细胞隔离和片外隔离将被开发并用于改善我们的NIH 原型。 AIM 3：高通量机械测试。为了实现高通量，需要修改的微流体和更快的检测技术。在此阶段，耦合将探索流体动力和光学力量以提高设备性能。此外，时变的光电将用于识别最佳信号响应和动态物理特性。