High-throughput Full Spectrum Cell Sorter

高通量全谱细胞分选仪

• 批准号: 10431645
• 负责人: Vinh Nguyen
• 金额: $ 75.24万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10431645
• 关键词: COVID-19; California; Cell Count; Cell Separation; Cells; Clinical; Clinical Research; Collection; Communities; Complement; Complex; Cost Savings; Coupled; Cytometry; Data; Devices; Dimensions; Disease Progression; Disease remission; Equipment; Faculty; Flow Cytometry; Funding; Maintenance; Paper; Peer Review; Performance; Pharmaceutical Preparations; Population; Preparation; Process; Publications; Publishing; Research; Research Personnel; Resolution; Sampling; San Francisco; Sorting; Technology; United States National Institutes of Health; Universities; data acquisition; experience; instrument; interest; next generation; tool

Project Summary / Abstract The University of California, San Francisco (UCSF) Parnassus Flow Cytometry CoLab (PFCC) is requesting funds to purchase a next generation Full Spectrum Cell Sorter. We have chosen the ThermoFisher Invitrogen™ BigFoot full spectrum cell sorter as the instrument that best fulfills the needs of the unsatiable demand for high resolution multidimension single cell data by our highly esteemed NIH-funded faculty. The BigFoot will complement our current arsenal of high-parameter analysis equipment such as the Fluidigm CyTOFs, Cytek Aurora and Becton Dickson FACSymphony X50 while most importantly adding the highly desired cell sorting capability. The BigFoot maintains the standard cell processing workflow PFCC users are accustomed to in conventional bandpass-based flow cytometers as well with the Aurora Full Spectrum Analyzer at PFCC. We will take advantage of the BigFoot's versatility and powerful spectral technology by enabling collection of fluorescent emission profiles through most of the visible spectra to near-infrared ranges making the number of parameters that can be investigated on this instrument approaching 35 to 40. The UCSF community has rapidly taken advantage of the availability of the full spectrum technology for their high-parameter research, the BigFoot will uniquely add cell sorting capabilities to aid the quest to understand the complex dynamics of changing cell population in disease progression, therapy and remission. Our Researchers have extensive experience in high-parameter data acquisition/analysis using Mass Cytometry resulting in 29 high impact publications over the last 4 years at UCSF, with at least 5 more in review. However, full spectrum flow cytometry is becoming a formidable technology to multiparameter analysis using mass cytometry; transition to the Aurora full spectrum analyzer has gained momentum due to the ease of sample preparations, simplicity of use, high-throughput (50 times faster) and especially the cost savings compared to mass cytometry. Within 1 year of installation, a peer review paper using the Aurora has been published. The high-throughput capabilities coupled with minimal sample loss during preparation and acquisition makes the technology amenable to samples with extremely low cell numbers. Hence, sorting on high-parameter panels of over 25 fluorescent markers has been in demand; however, no cell sorter is currently available at UCSF that allows panels of more than 18 fluorescent markers to be used. The BigFoot will augment the Full Spectrum Flow Cytometry portfolio at UCSF and the ability to simultaneously sort cell populations of interest which will be foundational in providing further information on cellular dynamics during multi-drug clinical research. Due to the COVID-19 lockdown, and limited accessibility at PFCC an ever- increasing number of processed clinical samples has been accumulating adding to the already pre-existing bank of samples ready for high-parameter analysis and sorting. At PFCC, the BigFoot will be a user-controlled device available 24/7, with minimal setup or daily maintenance requirements, and it will be a major high-throughput and high-performance tool in the Core. The BigFoot will help alleviate the bottleneck of machine availability continually experienced by our cell sorting and high-parameter researchers at UCSF.

项目摘要 /摘要 加利福尼亚大学旧金山（UCSF）Parnassus流式细胞仪COLAB（PFCC）要求资金 购买下一代全频谱分隔器。我们选择了Thermofisher Invitrogen™BigFoot全光谱 细胞分辨率是最能满足对高分辨率多维单一需求需求需求的仪器 我们受尊敬的NIH资助的教师的细胞数据。大脚怪将完成我们目前的高参数武器库 分析设备，例如流体细胞，Cytek Aurora和Becton Dickson Facsymphony X50，而大多数 重要的是添加高度期望的细胞分选能力。大脚怪维护标准单元格处理工作流程 PFCC用户也习惯于基于带通通道的流式细胞仪以及Aurora全光谱 PFCC分析仪。我们将通过启用大脚怪的多功能性和强大的光谱技术来利用 通过大多数可见光谱收集荧光排放曲线到近红外范围 可以在接近35至40的乐器上进行调查的参数。UCSF社区已迅速采用 全频谱技术可用于高参数研究的优势，Bigfoot将独特地添加 细胞分选能力，以帮助了解疾病中细胞种群改变的复杂动态 进展，治疗和缓解。我们的研究人员在高参数数据方面有丰富的经验 在过去的4年中，使用质量细胞仪的采集/分析，在UCSF上产生了29个高影响出版物，AT 至少有5个审查。但是，全光流式细胞仪已成为多参数的强大技术 使用质量细胞仪进行分析；过渡到Aurora全光谱分析仪由于易于 样品制剂，简单性，高通量（快50倍），尤其是与质量相比的成本节省 细胞仪。在安装后的1年内，已经发布了使用Aurora的同行评审论文。高通量 功能以及在准备和获取过程中的最小样本损失，使该技术可正常 细胞数量极低的样品。因此，对超过25个荧光标记的高参数面板进行排序 需求；但是，UCSF目前尚无细胞分辨率，该分类器可允许18个以上的荧光面板 要使用的标记。大脚怪将增加UCSF的全光流式细胞仪组合和能力 类似地，将感兴趣的细胞种群排序，这将是提供有关蜂窝信息的进一步信息的基础 多药临床研究期间的动态。由于COVID-19锁定，PFCC的可及性有限 - 越来越多的加工临床样本正在累积，将已经预先存在的样本添加到 准备进行高参数分析和分类。在PFCC，BigFoot将是24/7的用户控制设备， 最少的设置或每日维护要求，这将是主要的高通量和高性能工具 核。大脚怪将有助于减轻我们的细胞分类和不断经历的机器可用性的瓶颈 UCSF的高参数研究人员。