Celigo S Imaging Cytometer (200-BFFL-S)

7.项目总结 我们正在申请资金，以购买塞利戈S成像细胞仪，一种先进的多用途细胞成像 和定量仪器，以加强和加快金丝雀中心早期癌细胞培养研究 斯坦福大学癌症检测中心。准确的细胞培养研究是一个重要和必要的组成部分 几乎所有癌症的早期发现和治疗研究。然而，我们仍然进行细胞培养分析半 通过使用血细胞计数仪手动计数一小部分胰酶消化或悬浮细胞(10-20μL)，并 并外推计数以估计细胞总数。最基本的 这个过程的问题在于使用诸如外推之类的技术，这内在地引入了内部 样本可变性，而胰酶作用影响细胞生长和其他下游评估。这种方法 除了耗时限制精确度和吞吐量，这对提高 祝我们的学习取得成功。我们还使用基于荧光显微镜的分析方法对细胞进行成像和计数 多种应用，包括癌细胞的存活、增殖、凋亡和死亡、转染率、 蛋白质表达、干细胞特性、克隆形成、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并不是最优的，因为这些分析需要实时 显微镜下可视化，手工数字分析，收集最终结果。由于延长了时间 细胞在显微镜下的暴露时间，细胞暴露在影响细胞生长的次优条件下 偶尔会对表达的荧光记者进行光漂白。此外，建议的 市面上可买到的CLEGO细胞成像系统集成了先进的Brightfield和荧光技术 用于细胞计数和定量的成像。该系统的设计是为了从根本上提高精度 细胞培养分析时间。该系统对整个油井中的所有细胞进行计数，而不需要进行胰酶消化 需要复制多个井来监测细胞随时间的生长。激光扫描的光学和技术 系统允许快速井边到边沿均匀的细胞计数，提供高效和增强的细胞培养 学习。该系统配有五个通道(一个亮场通道和四个荧光通道)以同时执行 对多口井的测量。它还配备了用户友好但全面的数据采集和 分析软件包括几个为特定应用程序预先配置的软件。该系统及其集成的 功能和先进的软件工具，将发挥关键作用的所有用户在金丝雀中心进行 基于生物标志物的血液检测和分子成像检测定位方法的研究 通过体外和体内诊断早期癌症。因为我们有机会对仪器进行评估 通过在我们中心的演示，我们相信该仪器将对我们目前的 研究主要由美国国立卫生研究院资助，以促进高质量数据的获取。

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.