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: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=738551
• 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并监测其对不同刺激或药物的反应是必不可少的。许多目前使用的细胞分析仪器使用化学标记、复杂的设备和耗时的准备技术。使用介电分析的微流控器件是一种很有前途的替代方案，因为它们是无标签的，可以微型化，并且使用纳升液体体积。该研究计划的目标是开发集成了微波介电传感器和致动器的微流体设备，用于流动单细胞分析，利用这些设备研究细胞对不同类型压力的响应，开发新的细胞介电模型，并将细胞介电特性与生理联系起来。组织、细胞和其他生物介质具有独特的介电特性，可用于提供丰富的信息，如细胞的生长阶段，无论它是健康的还是癌症的，或者是否发生了药物诱导的变化。目前测量单电池介电性能的大多数方法只使用一个或两个频率，在低频下使用阻抗或介电泳法。通过测量完整的光谱并将其扩展到微波频率，可以获得许多额外的信息，在微波频率下，场可以穿透细胞内部，并可以检测到大分子的极化率。由于单电池体积小，其介电谱测量非常困难。我们将探索新的微波介电传感技术，在较宽的频谱范围内测量单个细胞，并利用该技术开发多参数细胞介电模型。微流体装置将用于研究细胞对饥饿、热休克和微波电穿孔(EP)的反应。饥饿诱导的细胞凋亡检测在生物制药、生物加工和生物医学研究中具有重要意义。细胞在饥饿状态下的介电变化将与细胞生理学有关，所开发的细胞模型将被用来理解整体悬浮介电测量。热休克和暴露于高强度电场(诱发EP)可以永久或暂时地使细胞膜通透性。在癌症治疗中，它被用于转染法、提高药物摄取率、诱导细胞凋亡，或者在极端情况下用于裂解。微流控设备将被开发用于应用并同时测量与这两个应激源相关的细胞的介电变化。特别令人感兴趣的是连续波微波EP，在那里我们将探索使用低强度场来诱导EP效应。这项研究的成果将是集成了刺激功能的新型微流控单细胞介电光谱工具，提供新的非侵入性、无标签的单细胞分析仪器。该研究计划将为跨学科环境中的高素质人员提供理论和实验培训。