Analytical Cytometry

摘要共享资源01：分析细胞计数（AC-SR） 为了提高我们对癌症病因学的理解，并开发出更好的治疗方法， 对于研究人员来说，有能力从复杂的细胞中常规分析和分离离散的细胞类型， 异质种群分析细胞计数共享资源（AC-SR）实现了这一重要功能， COHCCC成员通过提供定义和分离离散免疫系统细胞和癌细胞的服务 基于细胞外和细胞内蛋白表达，定量和分离表达荧光蛋白的细胞， 标记，并检查离散的细胞生长状态。 AC-SR的具体目标： 目标1.支持流式细胞术实验，用于细胞和小颗粒的分析。 目标二。维护细胞分选仪器并进行多参数分选。 目标3.为研究人员提供培训支持，以进行独立的流式细胞仪分析， 在AC-SR中排序。 AC-SR提供流式细胞仪分析和分选仪器（18台分析仪，7台分选仪）， COHCCC的研究目标。AC-SR维护分析仪器（7-BD LSRII/Fortessa，Cytek Aurora、BD-Symphony、3-ThermoFisher Attune NxT、2-ACEA Quanteons、1-ACEA Novocyte、2-BD C6和BC CyAn），其能够对所有COH位点进行不同程度的参数检测。流式细胞仪Fluidigm CyTOF- Helios和小型粒子分析仪（例如，外泌体和纳米颗粒; NanoSight NS 300）也是可用的。 值得注意的是，Aurora、Symphony和CyTOF仪器是上一个融资周期以来的新仪器。这些文书 允许高参数实验，能够对癌细胞、癌症 微环境和免疫反应。所有分析仪器均可通过以下方式操作 研究人员或SR工作人员。六台细胞分选仪（BD Aria分选仪）位于COH站点， 4个可由经过培训的最终用户自行操作。COHCCC最近购买了一个额外的封闭 系统细胞分选仪（Miltenyi Tyto），以协助分选CAR-T细胞产物和溶瘤病毒研究需求。 SR由癌症遗传学和表观遗传学系教授Jeremy Stark博士指导， 由高素质的工作人员提供支持，维护和操作设备，同时还提供培训， 希望独立操作AC-SR仪器的COHCCC研究人员。 2017-21年按收入百分比计算的会员利用率：总计93.6（20 MCBC，4.1 DCT，24 CI，45 HM，0.26 CCPS） 成员出版物：172，95，影响因子>10 支持的赠款：169总计（4 ACS，12 CIRM，8 DoD，6 LLS，18 NCI of 32 NIH（1 P01，88 R 01，5 U 01））

Abstract Shared Resource 01: Analytical Cytometry (AC-SR) To advance our understanding of cancer etiology and to develop improved therapeutic approaches, it is critical for researchers to have the ability to routinely analyze and isolate discrete cell types from complex heterogeneous populations. The Analytical Cytometry Shared Resource (AC-SR) fulfills this vital function for COHCCC Members by providing services to define and isolate discrete immune system cells and cancer cells based on extracellular and intracellular protein expression, to quantify and isolate cells that express fluorescent markers, and to examine discrete cell growth states. Specific Aims for AC-SR: Aim 1. Support flow cytometry experiments for the analysis of cells and small particles. Aim 2. Maintain instrumentation for cell sorting and perform multiparameter sorting. Aim 3. Provide training support for researchers to perform independent flow cytometry analysis and sorting within the AC-SR. The AC-SR provides flow cytometry analysis and sorting instrumentation (18 analyzers, seven sorters) to support the research goals of COHCCC. The AC-SR maintains analysis instruments (7-BD LSRII/Fortessa, Cytek Aurora, BD-Symphony, 3-ThermoFisher Attune NxT, 2-ACEA Quanteons, 1-ACEA Novocyte, 2-BD C6, and BC CyAn) that enable varying degrees of parameter detection for all COH sites. A mass cytometer, Fluidigm CyTOF- Helios, and a small particle analyzer (e.g., exosome and nanoparticle; NanoSight NS300) is also available. Notably, the Aurora, Symphony, and CyTOF instruments are new since the last funding cycle. These instruments allow for high parameter experiments, enabling highly detailed analysis and interrogation of cancer cells, cancer microenvironments, and immunological responses. All analysis instruments can be operated either by researchers or by SR staff. Six cell sorting instruments (BD Aria sorters) are located across COH sites of which 4 are available for self-operation by the trained end user. COHCCC has recently purchased an additional closed system cell sorter (Miltenyi Tyto) to assist with sorting of CAR-T cell products and oncolytic viral research needs. The SR is directed by Dr. Jeremy Stark, a Professor in the Department of Cancer Genetics and Epigenetics and supported by highly qualified staff that maintain and operate the equipment while also providing training to COHCCC researchers who wish to operate AC-SR instruments independently. Members Utilization by %Revenue 2017–21: 93.6 Total (20 MCBC, 4.1 DCT, 24 CI, 45 HM, 0.26 CCPS) Publications by Members: 172, 95 with Impact Factor >10 Grants Supported: 169Total (4 ACS, 12 CIRM, 8 DoD, 6 LLS, 18 NCI of 32NIH (1P01, 88R01, 5U01))