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

• 批准号: 7916769
• 负责人: CHARLES Dionisio EGGLETON
• 金额: $ 31.12万
• 依托单位: COLORADO SCHOOL OF MINES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-15 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7916769
• 关键词: Biological Assay; Cell Count; Cell Separation; Cell Survival; Cells; Chemicals; Collaborations; Coupling; Data; Detection; Development; Device Designs; Devices; Erythrocytes; Health; Image; Individual; Infection; Investigation; Label; Laboratories; Laboratory Research; Lasers; Lead; Life; Malaria; Measurement; Measures; Mechanics; Methods; Microfluidics; Modeling; Molecular; Monitor; National Institute of Allergy and Infectious Disease; Optical Methods; Optics; Parasites; Performance; Phase; Physiology; Property; Reagent; Research; Rheology; Scheme; Signal Transduction; Source; Speed; Stretching; Suspension substance; Suspensions; System; Techniques; Technology; Testing; Time; United States National Institutes of Health; Work; base; cell type; cost; high throughput analysis; high throughput screening; improved; instrument; instrumentation; optical traps; physical property; public health relevance; response; sensor; vector

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: We propose to develop new methods based on physical property measurement for the high-throughput analysis of cells. Such techniques that avoid the need for labels can be not only simpler and less expensive, they can be less harmful to the cell for applications where cell viability post-assaying is desirable.

描述（由申请人提供）：我们介绍了一种独特的基于微流体的方法，用于高通量非破坏性细胞测定，而无需特定的标记或试剂。基于使用施加的光学力测量静态和动态细胞机械特性，我们将以高速高通量的方式应用这种技术（称为“光学拉伸”）。迄今为止，光学拉伸仅用于小细胞数量;然而，高强度、微尺度激光源以及这些在动态微流体系统内的集成已经使我们提出的方法成为可能。在这种情况下，完全集成的光学传感器和机械拉伸器将用于识别并根据需要分离单细胞。一旦被鉴定，这样的靶向细胞然后可以在芯片上运输到装置内的培养室，或者分配到标准生物实验室仪器中用于芯片外分析。虽然有广泛的需求，我们提出的技术将使用疟疾寄生虫感染的红细胞作为靶细胞进行测试和开发。这项工作将与NIAID的疟疾和病媒研究实验室合作完成。我们的目标包括：目标1：机械性能检测和解释。我们将采用集成在微流控系统中的光学操作方法，用于无标记，非破坏性的细胞机械特性测量。建模方法将被开发用于解释施加的力/变形实验数据和设备设计。在这里，疟疾感染的红细胞将提供一个很好的模型目标，因为细胞刚度变化显着寄生虫发展。为了展示大大简化的设备设计和相关的易用性，我们将在一个活跃的NIH实验室安装一台仪器。目的2：用于细胞鉴定和分离的光学操作。我们将在微流控系统中集成光学方法，用于单细胞检测和操作。在这里，将开发用于芯片上细胞隔离和芯片外隔离的方法，并用于改进我们安装的NIH原型。目标3：高吞吐量机械测试。为了实现高通量，将需要改进的微流体和更快的检测技术。在这一阶段，将探索流体动力学和光学力的耦合，以提高器件性能。此外，将采用时变光学力来识别最佳信号响应和动态物理特性。公共卫生相关性：我们建议开发基于物理性质测量的新方法，用于细胞的高通量分析。避免对标记的需要的此类技术不仅可以更简单和更便宜，对于其中细胞活力后测定是期望的应用，它们对细胞的伤害可以更小。