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)：我们引入了一种独特的基于微流控技术的方法，用于高通量、非破坏性的细胞分析，而不需要特定的标记或试剂。在使用外加光学力测量静态和动态细胞力学特性的基础上，我们将以高速、高通量的方式应用这项技术(称为“光学拉伸”)。到目前为止，光学拉伸仅用于小细胞数量；然而，高强度、微尺度的激光光源以及这些光源在动态微流控系统中的集成使我们提出的方法成为可能。在这方面，将使用完全集成的基于光学的传感器和机械担架来识别并根据需要隔离单个细胞。一旦确定，这些靶细胞就可以在芯片上传输到设备内的培养室内，或者分配到标准的生物实验室仪器中进行芯片外分析。虽然有广泛的需求，但我们提出的技术将以疟疾寄生虫感染的红细胞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。我们将使用集成在微流控系统中的光学操作方法来进行无标记、非破坏性的细胞机械性能测量。将开发用于解释外加力/变形实验数据和用于装置设计的建模方法。在这里，感染疟疾的红细胞将提供一个很好的模型靶点，因为细胞硬度在寄生虫发育过程中会发生巨大变化。通过展示极大简化的设备设计和相关的易用性，我们将在NIH活跃的实验室中安装一台仪器。目的2：光学操作用于细胞鉴定和分离。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，我们将开发芯片内细胞隔离和芯片外隔离的方法，并将其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，将需要改进的微流控技术和更快的检测技术。在这一阶段，将探索流体动力和光学力的耦合，以提高器件的性能。此外，还将使用时变的光学力来识别最佳的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特性测量的新方法，用于细胞的高通量分析。这种不需要标记的技术不仅更简单、更便宜，而且对于需要细胞活力后检测的应用来说，它们对细胞的危害更小。