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 加州大学旧金山分校(UCSF)帕纳苏斯流式细胞术联合实验室(PFCC)正在申请资金 购买下一代全光谱细胞分类器。我们选择了ThermoFisher Invitgen™BigFoot全光谱 细胞分选仪作为最能满足人们对高分辨率多维单元化需求的仪器 由我们备受尊敬的NIH资助的教职员工提供的细胞数据。大脚怪将补充我们目前的高参数武器库 分析设备，如Fluidigm CyTOF，Cytek Aurora和Becton Dickson FAC Symphony X50，而大多数 重要的是，增加了高度期望的细胞分选功能。BigFoot维护标准的细胞处理工作流 PFCC用户习惯于使用传统的基于带通的流式细胞仪以及Aurora全光谱 PFCC的分析员。我们将利用大脚怪的多功能性和强大的光谱技术，通过启用 收集通过大部分可见光光谱到近红外范围的荧光发射轮廓，使数量 在这台仪器上可以研究的参数接近35到40。加州大学旧金山分校的社区迅速采取了 利用可用的全光谱技术进行高参数研究，大脚怪将独一无二地增加 细胞分选能力，有助于了解疾病中细胞群体变化的复杂动态 进展、治疗和缓解。我们的研究人员在高参数数据方面拥有丰富的经验 使用质量细胞术进行采集/分析，在过去4年中在加州大学旧金山分校出版了29本高影响力的出版物，其中 至少还有5个人在审查中。然而，全光谱流式细胞术正在成为一项强大的多参数检测技术 使用质量细胞仪进行分析；过渡到Aurora全频谱分析仪的势头因其易于 样品制备，使用简单，高通量(速度快50倍)，特别是与批量相比节省了成本 细胞学。在安装后的一年内，发表了一份使用Aurora的同行审查文件。高吞吐量 能力，加上在准备和获取过程中最小的样品损失，使该技术易于 细胞数目极低的样本。因此，对超过25种荧光标记的高参数面板进行分类 一直很受欢迎；然而，加州大学旧金山分校目前还没有可用的细胞分选器，它允许18个以上的荧光板 要使用的标记。大脚怪将增强加州大学旧金山分校的全光谱流式细胞术产品组合，并能够 同时对感兴趣的细胞群体进行分类，这将是提供有关细胞的进一步信息的基础 多药临床研究中的动态。由于新冠肺炎的封锁，以及PFCC有史以来最大的障碍 越来越多的经过处理的临床样本一直在积累，这增加了已经存在的样本库 准备好进行高参数分析和分类。在PFCC，大脚怪将是一种用户控制的设备，全天候可用，具有 最低限度的设置或日常维护要求，它将成为 核心。大脚怪将有助于缓解我们的细胞分选和 加州大学旧金山分校的高参数研究人员。