Flow Cytometry Shared Resources

项目总结：流式细胞术共享资源 西尔维斯特综合癌症中心（西尔维斯特）的流式细胞术共享资源（FCSR）是一个 最先进的设施，提供全面的分析流式细胞术和高速细胞分选服务 这对许多研究项目的成功进展至关重要。社研中心主任和工作人员的工作 共同提供专业咨询，分析设计和研究培训以及完整的细胞计数 分析和细胞分选服务，以支持研究计划，出版物和资助申请。的 FCSR的六台先进的荧光流式细胞仪（三台细胞分析仪和三台细胞分选仪） 促进广泛的复杂实验，检查正常和癌症的生物学特性 细胞FCSR能够同时分析多达19种不同的荧光 参数和两个激光散射参数（前向和侧向散射）的细胞表面和 复杂细胞混合物中的细胞内抗原检测。FCSR执行各种实验分析， 例如复杂免疫表型分析、DNA含量和细胞周期分析、细胞信号传导和活化 增殖和凋亡的分析和测量。此外，FCSR提供荧光激活的 细胞分选服务，使研究人员能够以高纯度和高速度纯化大量细胞和单细胞， 基于复杂的多参数表型。这三种细胞分选仪具有流动分选的潜力， 在生物安全下，将17个荧光参数放入原液管或多孔板中进行单细胞流克隆 二级条件。FCSR的三位经验丰富的技术人员进行细胞分选以及连续 所有仪器的质量控制评估。他们还提供全面的用户培训， 流式细胞仪实验，包括实验设计，荧光试剂板的选择，仪器 设置和操作、数据采集和数据分析。用户经过培训，能够 操作分析细胞仪本身，然后允许在过程中或外部独立地这样做 正常营业时间。用户通过在线仪器预订和发票访问该设施 系统2017年，FCSR为41名西尔维斯特成员提供了细胞计数服务（其中95%是 由同行评审资金支持），占FCSR总使用量的78%。

PROJECT SUMMARY: FLOW CYTOMETRY SHARED RESOURCE The Flow Cytometry Shared Resource (FCSR) at Sylvester Comprehensive Cancer Center (Sylvester) is a state-of-the-art facility offering comprehensive analytical flow cytometry and high-speed cell sorting services that are pivotal to the successful progress of many research projects. The FCSR Director and staff work collectively to provide professional consultation, assay design, and research training as well as full cytometric analysis and cell sorting services to support research programs, publications, and grant applications. The FCSR’s six sophisticated fluorescence flow cytometry instruments (three cell analyzers and three cell sorters) facilitate a wide range of complex experiments examining the biological properties of both normal and cancer cells. The FCSR has the capability to provide simultaneous analysis of up to 19 different fluorescent parameters and two laser light-scatter parameters (forward and side scatter) for both cell surface and intracellular antigen detection in complex cell mixtures. The FCSR performs a variety of experimental analyses, such as complex immunophenotyping, DNA content and cell cycle analysis, cell signaling and activation analysis, and measurement of proliferation and apoptosis. Further, the FCSR provides fluorescence-activated cell sorting services, which allow investigators to purify both bulk and single cells at high purity and high speed, based upon complex multiparametric phenotyping. The three cell sorters have the potential to flow-sort up to 17 fluorescence parameters into bulk tubes or into multi-well plates for single cell flow cloning under Biosafety Level 2 conditions. The FCSR’s three highly experienced technicians perform cell sorting as well as continuous quality control assessment for all instruments. They also provide comprehensive user training in all aspects of flow cytometric experiments, including experimental design, fluorescent reagent panel selection, instrument setup and operation, data acquisition, and data analysis. Users are trained such that they are capable of operating the analysis cytometers themselves and then are permitted to do so independently during or outside regular operating hours. Users access the facility through an online instrument reservation and invoicing system. In 2017, the FCSR provided cytometry services to 41 Sylvester members (95% of whom were supported by peer-reviewed funding), which comprised 78% of total FCSR usage.