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

使用具有计算特异性的相位成像进行无标记细胞活力测定

• 批准号: 10620330
• 负责人: CATALIN CHIRITESCU
• 金额: $ 64.99万
• 依托单位: PHI OPTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-10 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10620330
• 关键词: 3-Dimensional; Agitation; Agreement; Binding; Biological Assay; Bioreactors; Businesses; Cell Count; Cell Culture Techniques; Cell Cycle; Cell Line; Cell Survival; Cells; Cellular Assay; Chemicals; Classification; Computer software; Consumption; DNA; Data; Detection; Development; Dryness; Ensure; Escherichia coli; Feasibility Studies; Fluorescence; Funding; Geometry; Goals; Growth; Hour; Image; Imaging Device; Incubated; Investments; Label; Learning; Legal patent; Letters; Mammalian Cell; Marketing; Measurement; Mechanics; Membrane; Microscope; Microscopy; Modality; Modeling; Monitor; Morphology; Optical Instrument; Optics; Output; Performance; Pharmacologic Substance; Phase; Photobleaching; Phototoxicity; Pichia; Population; Procedures; Process; Production; Pump; Reagent; Recombinant Proteins; Research; Research Project Grants; Route; Saccharomyces; Sales; Sampling; Small Business Innovation Research Grant; Software Tools; Specialist; Specificity; Specimen; Stains; Stimulus; Stress; Structure; Suspensions; Technology; Testing; Time; Training; Trypan Blue; Trypsin; Work; artificial intelligence algorithm; cell growth; cell injury; cellular imaging; commercial prototype; commercialization; deep learning; deep learning algorithm; deep neural network; design; digital; experience; feeding; fluorescence imaging; fluorophore; high risk; imaging modality; imaging system; instrument; interest; manufacture; monolayer; novel; operation; prototype; quality assurance; quantitative imaging; research and development; software development; tomography

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.

项目总结/摘要 活细胞浓度是上游技术中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， 存在对药物生物反应器中的细胞活力的在线/在线监测的未满足的需求 实时产品质量保证，如过程分析技术（PAT）和质量 设计倡议。 与第一阶段工作相当的初步工作已经产生了包括流动池装置的细胞成像系统， 模拟在线搅拌生物反应器取样路线。还建立了用于检测的概念验证， 通过PICS（具有计算特异性的相位成像）使用数字染色对活/死测定进行分类。 这个小企业创新研究项目提出建立一个商业原型仪器，用于真实的 用于在线/在线生物反应器监测的时间、定量和高通量细胞活力评分， 制药研发应用的重要指标。在此过程中，QPI+PICS的标准操作 仪器将得到完善，自动软件检测和分类的活/死化验将 完成 在这个直接第二阶段的项目结束时，我们将提供一个商业原型-一个“按下一个按钮”的可行性 扫描仪内置上述功能。