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.

项目概要/摘要 加州大学旧金山分校 (UCSF) Parnassus 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选仪。我们选择了 ThermoFisher Invitrogen™ BigFoot 全谱 细胞分选仪作为最能满足对高分辨率多维单细胞无法满足的需求的仪器 细胞数据由我们备受尊敬的 NIH 资助的教师提供。 BigFoot 将补充我们当前的高参数库 分析设备，例如 Fluidigm CyTOF、Cytek Aurora 和 Becton Dickson FACSymphony X50，而大多数 重要的是增加了非常需要的细胞分选能力。 BigFoot 保持标准的细胞处理工作流程 PFCC 用户习惯使用传统的带通流式细胞仪以及 Aurora Full Spectrum PFCC 的分析仪。我们将通过启用 BigFoot 的多功能性和强大的光谱技术来利用 通过大部分可见光谱到近红外范围收集荧光发射曲线，使得 可以在该仪器上研究的参数接近 35 到 40。 UCSF 社区已迅速采取行动 利用全光谱技术进行高参数研究，BigFoot 将独特地添加 细胞分选能力有助于了解疾病中细胞群变化的复杂动态 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 使用质谱细胞术进行采集/分析，在过去 4 年中在 UCSF 发表了 29 篇高影响力出版物，其中 至少还有 5 个正在审核中。然而，全谱流式细胞术正在成为多参数分析的一项强大技术。 使用质谱流式分析仪进行分析；由于易于使用，向 Aurora 全光谱分析仪的过渡已获得动力 样品制备、使用简单、高通量（快 50 倍），尤其是与质量相比节省成本 细胞计数术。安装后一年内，发表了一篇使用 Aurora 的同行评审论文。高通量 能力加上制备和采集过程中最小的样品损失使得该技术适合 细胞数量极低的样本。因此，对超过 25 个荧光标记的高参数面板进行分类已成为可能 受到需求；然而，目前 UCSF 还没有提供允许超过 18 个荧光细胞组的细胞分选仪。 要使用的标记。 BigFoot 将增强 UCSF 的全谱流式细胞术产品组合，并增强 同时对感兴趣的细胞群进行分类，这将是提供有关细胞的进一步信息的基础 多药临床研究期间的动态。由于 COVID-19 封锁以及 PFCC 的访问受限， 已处理的临床样本数量不断增加，添加到已经存在的样本库中 为高参数分析和排序做好准备。在 PFCC，BigFoot 将是一款 24/7 全天候可用的用户控制设备， 最少的设置或日常维护要求，它将成为该领域的主要高通量和高性能工具 核。 BigFoot 将有助于缓解我们的细胞分选和生产过程中不断遇到的机器可用性瓶颈 加州大学旧金山分校的高参数研究人员。