High-throughput Full Spectrum Cell Sorter

项目总结/摘要 加州大学旧金山分校弗朗西斯科（UCSF）帕纳苏斯流式细胞术联合实验室（PFCC）正在申请资金， 购买下一代全频谱细胞分选仪。我们选择了ThermoFisher Invitrogen™ BigFoot全光谱 细胞分选仪作为最好的仪器，满足了高分辨率多维单细胞的不可满足的需求， 我们备受尊敬的国立卫生研究院资助的教师提供的细胞数据。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 FACSymphony X50，而大多数分析设备，如Fluidigm CyTOF、Cytek Aurora和Becton Dickson FACSymphony X50， 重要的是增加了高度期望的细胞分选能力。BigFoot维护标准单元处理工作流程 PFCC用户习惯于使用传统的带通流式细胞仪以及Aurora全谱流式细胞仪 PFCC的分析仪。我们将利用大脚怪的多功能性和强大的光谱技术， 通过大部分可见光谱到近红外范围收集荧光发射曲线， 可以在该仪器上研究的参数接近35至40。UCSF社区迅速采取了 利用全光谱技术进行高参数研究的优势，大脚将独特地增加 细胞分选能力，以帮助了解疾病中细胞群变化的复杂动力学 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 在过去的4年里，在UCSF使用质谱细胞术进行采集/分析，产生了29篇高影响力的出版物， 至少有5个在审查中。然而，全光谱流式细胞术正在成为一种强大的多参数技术， 使用质谱仪进行分析;由于易于 样品制备，使用简单，高通量（快50倍），特别是与大规模生产相比， 细胞仪在安装后的一年内，发表了一篇使用Aurora的同行评审论文。高通量 在制备和获取过程中，样品损失最小， 细胞数量极低的样本。因此，在超过25个荧光标记物的高参数组上进行分选， 然而，目前在UCSF没有细胞分选仪可以允许超过18个荧光标记的面板。 要使用的标记。BigFoot将增强UCSF的全谱流式细胞术产品组合， 同时分选感兴趣的细胞群体，这将是提供细胞生物学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，增加了已经存在的样本库 准备进行高参数分析和分选。在PFCC，大脚将是一个用户控制的设备，可24/7， 最小的设置或日常维护要求，它将是一个主要的高吞吐量和高性能的工具， 核心BigFoot将有助于缓解我们的细胞分选持续经历的机器可用性瓶颈， 加州大学旧金山分校的高参数研究人员

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.