Correlation of imaging and impedimetric data to predict cell state and changes in bioreactors

成像和阻抗测量数据的相关性以预测细胞状态和生物反应器的变化

• 批准号: 2439724
• 金额: --
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2439724
• 关键词: Correlation; imaging; impedimetric; data; predict

The ability to monitor and influence the physiological state of a cell (population) is of critical importance not just for reproducible results in academia but especially in the pharmaceutical industry. Here, the cell physiological state is directly correlated with the production of biologicals and consequently profitability and quality.There are multiple ways to measure the cell state but most look at a single parameter. It is much more informative to integrate and correlate different types of data. Correlative Light Electron Microscopy (CLEM) is one of the most powerful imaging technologies combining the advantages of (live) light microscopy (LM) with the nanometer spatial resolution of electron microscopy (EM) into one experiment. Using this technology, key biological questions have been answered [1].Cell structures and metabolic activity affect the electrical properties of the cell and by employing a broadband approach to impedance spectroscopy multiple cell properties can be investigated. In general terms, high-frequencies are associated with changes in the cell cytoplasm/internal structures (10-60Mz), middle-frequencies with the cell membrane (2-10MHz) and lower-frequencies with cell size (0.1-2 MHz). We have shown that dose-dependent characteristics of the impedance spectra of cell culture when exposed to a toxic challenge are related to the toxin's mode of action [2].There is a need and drive to extract more than just LM and EM data from a single experiment and to include other types of information, expanding the CLEM field to an approach called Correlative Multimodal Imaging (CMI). In principle the CMI approach would extract any type of information from a single event and correlate and integrate the data.This project enables, for the first time, to understand and correlate morphological and anatomical changes in the cell structure, detected microscopically, to changes in characteristics of an impedance spectrum. Impedance spectroscopy can be employed to monitor cells growing in a bioreactor and by understanding the changes in the impedance spectra related to cell stress will enable the early detection and mitigation of cell stress.We propose to develop a CMI approach that combines live imaging with impedance measurements in control and stressed (e.g. toxins) conditions in order to better understand the different responses to stressors. We will use live calcium and / or ATP imaging in combination with impedance measurements. Subsequently we will also aim to introduce stress receptor-GFP imaging and EM morphology into this correlative approach to increase the power of the technology.

监测和影响细胞(群体)的生理状态的能力不仅对学术界，尤其是制药业的可重复性结果至关重要。在这里，细胞的生理状态与生物制品的生产直接相关，从而与利润和质量相关。有多种方法来衡量细胞状态，但大多数方法只看一个参数。集成和关联不同类型的数据会提供更多的信息。相关光电子显微镜(CLEM)是将活光学显微镜(LM)的优点和电子显微镜(EM)的纳米空间分辨率结合在一起的最强大的成像技术之一。利用这项技术，关键的生物学问题得到了解答[1]。细胞结构和代谢活动影响细胞的电学性质，通过采用宽带方法进行阻抗频谱分析，可以研究多种细胞性质。一般来说，高频与细胞质/内部结构的变化(10-60 Mz)有关，中频与细胞膜有关(2-10 MHz)，低频与细胞大小有关(0.1-2 MHz)。我们已经证明，细胞培养在受到毒物攻击时阻抗谱的剂量依赖特性与毒素的作用模式有关[2]。我们需要和推动从单个实验中提取更多的LM和EM数据，并包括其他类型的信息，将Clem领域扩展到一种称为相关多模式成像(CMI)的方法。原则上，CMI方法将从单个事件中提取任何类型的信息，并将数据关联和整合。该项目首次使通过显微镜检测到的细胞结构的形态和解剖变化与阻抗谱特征的变化相关联。阻抗谱可以用来监测细胞在生物反应器中的生长，通过了解与细胞应激相关的阻抗谱的变化，可以及早发现和缓解细胞应激。我们建议开发一种CMI方法，将现场成像与控制和应激(如毒素)条件下的阻抗测量相结合，以更好地了解应激源的不同反应。我们将结合阻抗测量使用实时钙和/或ATP成像。随后，我们还将致力于将应激受体-GFP成像和EM形态学引入到这种相关的方法中，以增加该技术的威力。