CCR-Frederick Flow Cytometry Core

CCR-Frederick 流式细胞术核心

• 批准号: 8350148
• 负责人: Kathleen Noer
• 金额: $ 86.79万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8350148
• 关键词: Accounting; Antibodies; Basic Science; Cell Line; Cells; Color; Complex; Data; Data Analyses; Flow Cytometry; Fluorescence; Future; Genetic Transcription; Goals; Government; Individual; Inflammation; Inflammatory Response; Laboratory Scientists; Lasers; Leukocytes; Lung Neoplasms; Malignant Neoplasms; Measures; Membrane Lipids; Modeling; NCI Center for Cancer Research; National Cancer Institute; Paper; Population; Principal Investigator; Productivity; Publishing; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Role; Running; Sampling; Science; Services; Side; Sorting - Cell Movement; Staining method; Stains; Stem cells; Techniques; Technology; Time; Training; Transgenic Mice; Tumor Stem Cells; Tumor-Infiltrating Lymphocytes; Update; Work; anticancer research; cell type; cellular transduction; design; detector; experience; instrument; instrumentation; investigator training; meetings; nile red; novel; programs; red fluorescent protein; research study; tool; tumor; tumor progression

The fast changing field of flow cytometry has required the flow core to replace old instrumentation and up-grade the newer instruments that we have in order to meet the requirements of our CCR investigators. The legacy MoFlo cell sorter was transfered to NEI in order for us to obtain a BD FACSAria II with a 100mW UV laser to enable core to sorttumor stem cells using the side population technique. The green lasers that were added to an anlyzer and a cell sorter is being used by numerous investigators to detect red fluorescent proteins expressed in transgenic mice and transfected or transduced cell lines. The green laser has also been used to sort lung tumor stem cells from the bulk of the tumor using the differing expression of nile red in the membrane lipids of the two cell types. More of the investigators have increased the complexity of their experiments and are routinely running experiments with 6-9 antibodies to define distinct populations of tumor infiltrating lymphocytes in different tumor models. Several linvestigators are now sorting tumor infiltrating leukocytes from various tumors in order to detect RNA expression involved in the inflammatory response to the tumor measured by quantitative RT-PCR. The larger number of fluorescence detectors has greatly increased the quality and complexity of information generated about the role of inflammation in tumor progression with each experiment. The flow core staff has trained seven investigators in the first three quarters of the year to run their own samples. The instruments are used daily by the trained investigators often into the night and on the weekends. This leaves more time for the flow core staff to sort approximately 537 samples and analyze about 6700 samples in the first three quarters of this year. Without the investigators acquiring the data and analyzing their own samples, the government would have to hire full time experienced individuals to perform the same volume of work. At least 23 papers have been published in the past year using data obtained in the flow core. In the past year, data has been generated (or samples have been sorted) for 78 individual scientists from the laboratories of 36 principal investigators. Although the CCR-Frederick Flow Cytometry Core is part of the CIP, investigators from this program account for approximately 49% of the use of this facility. The future goals include keeping up with cutting-edge technology in the flow lab by expanding the pool of investigators trained to run and analyze their own samples to all of CCR in Frederick thereby expanding the use, productivity and quality of the science generated using the core's instrumentation and expertise. This could be achieved by adding a 405 laser to the LSRII Fortessa and replacing the two BD FACScans with small 2 laser 6-color instruments.

为了满足CCR研究人员的要求，流式细胞术领域的快速变化要求流式核心取代旧仪器并升级我们现有的新仪器。传统的MoFlo细胞分选器被转移到NEI，以便我们获得带有100mW紫外激光的BD FACSAria II，使核心能够使用侧群技术对肿瘤干细胞进行分选。添加到分析仪和细胞分选器中的绿色激光被许多研究人员用于检测转基因小鼠和转染或转导细胞系中表达的红色荧光蛋白。利用两种细胞类型膜脂中尼罗红的不同表达，绿色激光也被用于从大部分肿瘤中分选肺肿瘤干细胞。越来越多的研究人员增加了实验的复杂性，并常规地使用6-9抗体进行实验，以确定不同肿瘤模型中肿瘤浸润淋巴细胞的不同群体。目前，一些研究人员正在从各种肿瘤中分选浸润性白细胞，以便通过定量RT-PCR检测参与肿瘤炎症反应的RNA表达。荧光探测器数量的增加大大提高了每次实验中关于炎症在肿瘤进展中的作用的信息的质量和复杂性。今年前三个季度，流动核心人员培训了7名调查人员，以运行他们自己的样本。这些仪器每天都由训练有素的调查人员使用，经常使用到深夜和周末。这使得流量核心人员有更多的时间在今年前三季度对大约537个样品进行分类，并对大约6700个样品进行分析。如果没有调查人员获取数据并分析他们自己的样本，政府将不得不聘请有经验的全职人员来完成同样数量的工作。在过去的一年里，至少有23篇论文使用了从流动岩心中获得的数据。在过去的一年里，已经为来自36个主要研究人员的实验室的78名科学家生成了数据（或整理了样本）。虽然CCR-Frederick流式细胞仪核心是CIP的一部分，但来自该项目的研究人员约占该设施使用的49%。未来的目标包括通过扩大经过培训的研究人员的数量，以运行和分析弗雷德里克所有CCR的自己的样品，从而扩大使用核心仪器和专业知识产生的科学的使用，生产力和质量，从而跟上流动实验室的尖端技术。这可以通过在LSRII Fortessa上增加一台405激光器和用小型2激光6色仪器替换两台BD facscan来实现。