High throughput and accuracy single cell dielectric analysis and dielectric dyes

测量电池的介电性能在研究、生物医学和工业领域有着重要的应用。例如，生物制药公司Biogen在其所有制造过程中，使用批量介电分析来控制哺乳动物细胞培养物的喂养，最大限度地提高了药物生产率。由于细胞和细胞群体的不均匀介电性质，整体介电分析具有局限性。单电池介电分析在确定悬浮液中电池的真实介电性能方面要优越得多。我的计划将集中在：1)开发一种集吞吐量、准确度、分析时间和频率范围于一体的光学过渡介电(DEP)细胞仪。光学过境DEP细胞仪分析来自细胞的过境光曲线的信号，类似于行星在恒星前面经过，以测量粒子速度并推断介电性质。使用差分信号方法将提高精度。电极将被设计成允许分析频率高达500兆赫，以收集关于细胞核和细胞器的信息。通道和检测器的几何形状将被设计为显著增加吞吐量和减少分析时间。每个细胞的吞吐量超过1,000个，能够在10‘秒内捕获多频谱S。2)开发反映细胞生理状态的独特介电染料。介电染料是一种化合物，与未处理的细胞相比，产生可测量的介电对比度，这取决于特定的细胞属性(例如，生化过程)。例如，细胞暴露于低聚阿霉素会抑制线粒体ATP的产生。较低的ATP会减慢膜离子泵的速度，并导致细胞质电导率的下降，这可以用来估计线粒体ATP的产生水平。我建议测试一些最有希望的候选者，使用时间拟合来双壳层介电模型。这项独特的研究将导致单电池介电分析的新模式。这项研究将提供一种新的单电池介电分析工具，具有多种特性，包括简单的组件，使其成为单电池介电分析的首选方法之一。介电染料有可能扩大可用单电池介电分析进行的分析类型。例如，对线粒体ATP进行简单的检测将在生物制药控制和癌症检测中得到应用，在癌症检测中，线粒体ATP的缺乏被认为是癌症的标志。光学传输细胞仪单细胞介电分析将最大限度地提高哺乳动物细胞培养的生产率，从而使加拿大的生物制药行业，如急诊、治疗、赛诺菲等受益。有了HQP和该项目的技术，Micralyne和Dalsa等微流控仪器公司将能够提供新的产品和市场。

Measuring the dielectric properties of cells has important applications in research, biomedical and industrial sectors. For example, in all its manufacturing the biopharma company Biogen maximizes drug productivity using bulk dielectric analysis to control feeding of mammalian cell cultures. Bulk dielectric analysis has limitations due to the inhomogeneous dielectric nature of cells and cell populations. Single cell dielectric analysis is far superior for determining the true dielectric properties of cells in suspension. My program will focus on: 1) Development of an optical transit dielectrophoresis (DEP) cytometer with a highly beneficial combination of throughput, accuracy, analysis time and frequency range. The optical transit DEP cytometer analyzes signals from transit light curves from cells, similar to planets passing in front of a star, to measure particle velocity and infer dielectric properties. Accuracy will be improved using differential signals approaches. Electrodes will be designed to allow analysis frequencies up to 500 MHz, to glean information on cell nuclei and organelles. The channel and detector geometry will be designed to dramatically increase throughput and reduce analysis time. Throughputs of greater than 1000 cells per and the ability to capture mutli-frequency spectra in 10's of seconds. 2) Development of unique dielectric dyes of cellular physiological status. A dielectric dye is a compound that produces a measurable dielectric contrast compared to untreated cells that is dependent on a particular cell property (e.g. biochemical process). For example, exposing a cell to Oligamycin inhibits mitochondrial ATP production. Lower ATP slows membrane ion pumps and causes a drop in cytoplasm conductivity that can be used to estimate the level of mitochondrial ATP production. I propose to test some of the most promising candidates using temporal fits to double shell dielectric models. This unique research will result in new modalities of single cell dielectric analysis. This research will provide a new single cell dielectric analysis tool with a combination of properties, including simple components, that make it a very promising candidate to become one of the preferred methods for single cell dielectric analysis. The dielectric dyes have the potential to expand the types of assays that can be carried out using single cell dielectric analysis. For example, having a simple assay for mitochondrial ATP would have application in biopharma control and for cancer detection where the lack of mitochondrial ATP is considered a hallmark of cancer. The optical transit cytometer single cell dielectric analysis will benefit the Canadian biopharma industries such as Emergent, Therapure, Sanofi by maximizing the productivity of the mammalian cell cultures. With the HQP and technology from this program microfluidics instrumentation companies such as Micralyne and Dalsa will be able to offer new products and markets.