Celigo S Imaging Cytometer (200-BFFL-S)

Celigo S 成像细胞仪 (200-BFFL-S)

• 批准号: 9493316
• 负责人: Ramasamy Paulmurugan
• 金额: $ 17.37万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9493316
• 关键词: Affect; Aliquot; Apoptosis; Biological Assay; Blood Tests; Cell Count; Cell Culture Techniques; Cell Survival; Cells; Cessation of life; Computer software; Data Analyses; Data Quality; Diagnostic; Dose; Early treatment; Exposure to; Fluorescence Microscopy; Funding; Image; Imagery; In Vitro; Malignant Neoplasms; Manuals; Measurement; Methods; Microscope; Monitor; Optics; Pharmaceutical Preparations; Photobleaching; Play; Process; Property; Reporter; Research; Sampling; Scanning; Screening for cancer; Serinus; Software Tools; Stem cells; System; Techniques; Technology; Time; Transfection; Treatment Efficacy; United States National Institutes of Health; base; blood-based biomarker; cancer cell; cell growth; cellular imaging; cytotoxicity; data acquisition; design; digital; efficacy study; fluorescence imaging; imaging approach; imaging system; improved; in vivo; instrument; molecular imaging; protein expression; response; success; user-friendly

7. PROJECT SUMMARY We are requesting funding to purchase Celigo S Imaging Cytometer, an advanced multi-purpose cell imaging and quantitation instrument, to enhance and expedite cancer cell culture studies at the Canary Center for Early Cancer Detection at Stanford. Accurate cell culture studies are an important and necessary component of nearly all cancer early detection and therapy studies. However, we still perform cell culture analysis semi- manually by counting a small aliquot of trypsinized or suspended cells (10-20 μl) using a hemocytometer and brightfield microscope and extrapolating the count to estimate the total number of cells. The fundamental problem of this process lies on the use of techniques such as extrapolation, which inherently introduces intra- sample variability while trypsinization affecting cell growth and other downstream assessments. This method beyond being time consuming limit the accuracy and throughput, which are important for improving the success of our studies. We also use fluorescence microscopy-based assays to image and count cells for various applications including cancer cell viability, proliferation, apoptosis and death, transfection efficiency, protein expression, stem cell properties, colony formation, cytotoxicity, drug dose/response and treatment efficacy studies. The current methods are not optimal for our research, as these assays require real time visualization under microscope and manual digital analysis for collecting final results. Due to prolonged exposure time of cells under microscope, the cells are exposed to sub-optimal conditions that affect cell growth with occasional photobleaching of the expressed fluorescent reporters. Alternatively, the proposed commercially available Celigo cell imaging system integrates both advanced brightfield and fluorescent imaging for cell counting and quantification. The system is designed to fundamentally improve the accuracy and time of cell culture analysis. The system counts all cells in the entire well without trypsinization avoiding the need of replicate multiple wells to monitor cell growth over time. The optics and scanning technology of the system allow for fast well edge-to-edge uniform cell counting providing efficient and enhanced cell culture studies. The system comes with five channels (one brightfield and four fluorescent) to simultaneously perform measurements over multiple wells. It also comes with user-friendly but comprehensive data acquisition and analysis software including several preconfigured for specific applications. The system with its integrated functionality and advanced software tool, will play a key role for all users at the Canary Center conducting research on developing biomarker based blood tests and molecular imaging approaches to detect and localize early cancers through both in vitro and in vivo diagnostics. As we had a chance to evaluate the instrument through a demo at our center, we are confident that this instrument will provide significant impact to our current research mostly NIH funded by facilitating acquisition of high quality data.

7.项目摘要 我们正在申请资金购买Celigo S成像细胞仪，这是一种先进的多用途细胞成像仪。 和定量仪器，以加强和加快癌细胞培养研究在金丝雀中心的早期 斯坦福大学的癌症检测。准确的细胞培养研究是一个重要和必要的组成部分， 几乎所有的癌症早期检测和治疗研究。然而，我们仍然进行细胞培养分析， 通过使用血细胞计数器手动计数一小等份胰蛋白酶化或悬浮细胞（10-20 μl）， 明视野显微镜下观察细胞并外推计数以估计细胞总数。根本 该过程的问题在于使用诸如外推的技术，该技术固有地引入了内插， 胰蛋白酶消化影响细胞生长和其他下游评估时的样品变异性。该方法 除了耗时之外，还限制了准确度和吞吐量，这对于提高 我们的研究的成功。我们还使用基于荧光显微镜的分析来成像和计数细胞， 各种应用包括癌细胞活力、增殖、凋亡和死亡，转染效率， 蛋白质表达、干细胞特性、集落形成、细胞毒性、药物剂量/反应和治疗 功效研究。目前的方法对于我们的研究来说不是最佳的，因为这些测定需要真实的时间 显微镜下可视化和手动数字分析，以收集最终结果。由于长时间 细胞在显微镜下的暴露时间，细胞暴露于影响细胞生长的次优条件下 表达的荧光报道分子偶尔会发生光漂白。另外，建议 市售的Celigo细胞成像系统集成了先进的明场和荧光 用于细胞计数和定量的成像。该系统旨在从根本上提高 和细胞培养分析的时间。系统计数整个孔中的所有细胞，而不进行胰蛋白酶消化， 需要复制多个威尔斯孔以监测细胞随时间的生长。光学和扫描技术， 系统允许快速孔边到边均匀细胞计数，提供有效和增强的细胞培养 问题研究该系统配有五个通道（一个明场和四个荧光），可同时执行 多个威尔斯井的测量。它还具有用户友好但全面的数据采集功能， 分析软件，包括几个预先配置的特定应用程序。该系统及其集成 功能和先进的软件工具，将发挥关键作用，为所有用户在金丝雀中心进行 研究开发基于生物标志物的血液检测和分子成像方法，以检测和定位 通过体外和体内诊断早期癌症。当我们有机会评估仪器时 通过在我们中心的演示，我们相信，该仪器将为我们目前的 主要由NIH资助的研究促进了高质量数据的获取。