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: 10484781
• 负责人: CATALIN CHIRITESCU
• 金额: $ 89.43万
• 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/10484781
• 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; Data Analyses; Detection; Development; Ensure; Escherichia coli; Feasibility Studies; Fluorescence; Funding; Geometry; Goals; Growth; Hour; Image; Imaging Device; Investments; Label; Learning; Legal patent; Letters; Liquid substance; Mammalian Cell; 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; Staining and Labeling; Stains; Stimulus; Stress; Structure; Suspensions; Technology; Testing; Time; Training; Trypan Blue; Work; artificial intelligence algorithm; base; cell growth; cell injury; cellular imaging; deep learning; deep learning algorithm; deep neural network; design; digital; experience; feeding; fluorescence imaging; fluorophore; high risk; imaging modality; imaging system; instrument; interest; 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.

项目概要/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 与生物制药高管和研发专家进行的客户发现讨论显示。 在合规性的驱动下，对制药生物反应器中的细胞活力进行在线/在线监测的需求尚未得到满足 具有实时产品质量保证，如过程分析技术 (PAT) 和质量 设计建议。 相当于第一阶段工作的初步工作已经产生了一个细胞成像系统，包括一个流动池设置， 还建立了模拟在线搅拌生物反应器采样路线以进行检测和验证。 通过 PICS（具有计算特异性的相位成像）使用数字染色对活/死检测进行分类。 这个小企业创新研究项目建议建立一个真正的商业原型仪器 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，满足 在此过程中，QPI+PICS 的标准操作成为制药研发应用感兴趣的指标。 仪器将得到改进，自动软件检测和活/死分析的分类将得到改进。 完全的。 在这个直接第二阶段项目结束时，我们将交付一个商业原型——“一键式”可行性 内置上述功能的扫描仪。