Label-free cell viability assays using Phase Imaging with Computational Specificity

项目摘要/摘要 在上游技术中，活细胞密度是最重要的关键性能指标之一 在哺乳动物细胞培养中。与生物制药高管和研发专家的客户发现讨论显示 在顺应性驱动的制药生物反应器中，对细胞活性的在线/在线监测的需求尚未得到满足 具有过程分析技术(PAT)中所述的实时产品质量保证，并通过 设计计划。 相当于第一阶段工作的初步工作已经导致了包括流动池装置的细胞成像系统，该流动池装置 模拟了在线搅拌生物反应器的采样路线。还建立了概念验证以用于检测和 通过PICS(具有计算特异性的相位成像)使用数字染色对活/死检测进行分类。 该小型企业创新研究项目建议为REAL构建一个商业原型工具 时间、定量和高通量细胞活性评分，用于在线/在线生物反应器监控，满足 对制药研发应用感兴趣的指标。在这一努力中，QPI+PICS的标准操作 仪器将得到改进，活/死分析的自动化软件检测和分类将 完成。 在这个直接的第二阶段项目结束时，我们将交付一个商业原型--一键操作的可行性 内置上述功能的扫描仪。

Project Summary/Abstract Viable cell concentration is one of the most important key performance indicator during upstream technologies in mammalian cell culture. Customer Discovery discussions with biopharma execs and R&D specialists revealed there is an unmet need for on-line/in-line monitoring of cell viability in pharma bioreactors driven by compliance with real-time product quality assurance as described in Process Analytical Technology (PAT) and Quality by Design initiatives. Preliminary work equivalent to Phase I effort has resulted in a cell imaging system including a flow cell setup that simulates an on-line stirring bioreactor sampling route. Proof-of-concept was also established for detection and classification of live/dead assays using digital staining via PICS (Phase Imaging with Computational Specificity). This Small Business Innovation Research project proposes to build a commercial prototype instrument for real time, quantitative and high-throughput cell viability scoring for on-line/in-line bioreactor monitoring that meets the metrics of interest for pharma R&D applications. During this effort the standard operation of the QPI+PICS instrument will be refined and the automated software detection and classification of live/dead assays will be completed. At the end of this Direct Phase II project we will deliver a commercial prototype -a “push-of-a-button” viability scanner with the above capabilities built in.