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.

项目摘要/摘要 可行的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揭示了 在线/在线监测由合规性驱动的药物生物反应器中的细胞生存力的需求未满足 如过程分析技术（PAT）和质量所述的实时产品质量保证 设计计划。 等同于第一期的初步工作导致了一个单元成像系统，包括流动池设置 模拟在线搅拌生物反应器采样途径。还建立了概念证明供检测和 使用图片数字染色（具有计算特异性的阶段成像）对活/死测定进行分类。 这个小型企业创新研究项目的提案，旨在建立一个现实的商业原型工具 时间，定量和高通量细胞的生存能力评分，用于在线/在线生物反应器监视 药物研发应用的兴趣指标。在这项工作中，QPI+图片的标准操作 仪器将进行完善，自动化软件检测和生命/死亡分析的分类将是 完全的。 在此直接第二阶段项目的结尾，我们将提供商业原型-a“按钮推动”的生存能力 具有上述功能内置的扫描仪。