Microwave Devices for Biosensors and Single Cell Dielectric Spectroscopy

用于生物传感器和单细胞介电谱的微波设备

• 批准号: RGPIN-2019-05859
• 负责人: Bridges, Greg
• 金额: $ 3.35万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=752047
• 关键词: Microwave; Devices; Biosensors; Single; Cell

Studying the physiology of biological cells and monitoring their response to different stimuli or drugs is essential in bioprocess and biomedical research. Many currently used instruments for cellular analysis utilize chemical markers, sophisticated equipment and time-consuming preparation techniques. Microfluidic devices employing dielectric analysis are a promising alternative as they are label-free, can be miniaturized and use nanoliter fluid volumes. The objectives of the proposed research program is to develop microfluidics devices integrated with microwave dielectric sensors and actuators for in-flow single cell analysis, use these to study cell response to different types of stress, develop new cell dielectric models, and relate cell dielectric properties to physiology. Tissues, cells and other biomedia have unique dielectric properties that can be used to provide a wealth of information, such as a cell's growth phase, whether it is healthy or cancerous, or if drug-induced changes have occurred. Most current methods for measuring single cell dielectric properties employ impedance or dielectrophoresis at low frequencies using only one or two frequencies. Much additional information can be gained by measuring the complete spectrum and extending this to microwave frequencies, a regime where fields can penetrate the cell interior and the polarizability of large molecules can be detected. Dielectric spectroscopy of single cells is extremely difficult due to their small size. We will explore new microwave dielectrophoresis sensing techniques to measure single cells over a wide frequency spectrum and use this to develop multi-parameter cell dielectric models. Microfliudic devices will be used to study cell response to starvation, heat shock and microwave electroporation (EP). Detection of starvation-induced apoptosis is important in biopharmaceutical bioprocessing and biomedical research. Dielectric changes to cells under starvation will be related to cell physiology and the developed cell models will be used to understand bulk-suspension dielectric measurements. Heat shock and exposure to high-intensity electric fields (inducing EP) can permanently or temporarily permeabilize cell membranes. This is employed for transfection, enhancing drug uptake in cancer therapy, inducing apoptosis, or in the extreme case, lysis. Microfluidic devices will be developed to apply and simultaneously measure the dielectric changes to cells associated with these two stressors. Of particular interest is CW microwave EP, where we will explore use of low-intensity fields to induce EP effects. The outcomes of the research will be new microfluidic single cell dielectric spectroscopy tools integrated with stimulus capabilities, providing new non-invasive label-free single cell analysis instruments. The research program will provide both theoretical and experimental training to highly qualified personnel in an interdisciplinary environment.

在生物过程和生物医学研究中，研究生物细胞的生理学并监测其对不同刺激或药物的反应至关重要。许多目前使用的仪器用于细胞分析，利用化学标记，复杂的设备和耗时的准备技术。使用介电分析的微流体设备是一种有前途的选择，因为它们不含标签，可以小型化并使用纳米液液体积。拟议的研究计划的目标是开发与微波介电传感器和执行器集成的微流体设备进行流量单细胞分析，使用这些设备来研究对不同类型的压力的细胞响应，开发新的细胞介电模型，并将细胞介电模型与生理学相关。组织，细胞和其他生物膜具有独特的介电特性，可用于提供大量信息，例如细胞的生长阶段，无论是健康的还是癌性的，或者发生了药物诱导的变化。测量单细胞介电特性的大多数当前方法仅使用一个或两个频率在低频下采用阻抗或介电性。可以通过测量完整的频谱并将其扩展到微波频率来获得许多其他信息，该机制可以穿透细胞内部，并且可以检测到大分子的极化性。由于其小尺寸，单细胞的介电光谱非常困难。我们将探索新的微波电介胶感应技术，以测量广泛的频谱上的单个细胞，并使用它来开发多参数细胞介电模型。 微升状设备将用于研究细胞对饥饿，热休克和微波电穿孔（EP）的反应。检测饥饿诱导的凋亡在生物药物生物处理和生物医学研究中很重要。饥饿下细胞的介电变化将与细胞生理有关，并且开发的细胞模型将用于了解散装悬浮介质测量。热休克和暴露于高强度电场（诱导EP）可以永久或暂时渗透细胞膜。这是用于转染，增强癌症治疗中的药物摄取，诱导凋亡或极端情况下的裂解。将开发微流体设备以应用和同时测量与这两个应激源相关的细胞的介电变化。 CW微波EP尤其令人感兴趣，我们将在其中探索使用低强度场来诱导EP效应的使用。该研究的结果将是与刺激能力集成的新的微流体单细胞介电光谱工具，从而提供了新的非侵入性无标签单细胞分析仪器。该研究计划将在跨学科环境中为高素质的人员提供理论和实验培训。