CCR-Frederick Flow Cytometry Core

快速变化的流式细胞仪领域要求流动核心更换旧仪器，并升级我们拥有的较新仪器，以满足我们CCR调查人员的要求。传统的MoFlo细胞分选机被转移到NEI，以便我们获得带有100 mW紫外线激光的BD FACSAria II，使CORE能够使用侧群技术对肿瘤干细胞进行分选。许多研究人员正在使用添加到分析仪和细胞分选器中的绿色激光来检测在转基因小鼠和转基因或转导的细胞系中表达的红色荧光蛋白。利用尼罗红在两种细胞类型的膜脂中的不同表达，绿色激光也被用于从肿瘤的大部分中分离出肺肿瘤干细胞。更多的研究人员增加了他们实验的复杂性，并常规地使用6-9个抗体进行实验，以确定不同肿瘤模型中肿瘤浸润性淋巴细胞的不同群体。一些研究人员现在正在对各种肿瘤中的肿瘤浸润性白细胞进行分类，以检测参与肿瘤炎症反应的RNA的表达，这是通过定量RT-PCR来测量的。更多的荧光探测器大大增加了每次实验产生的关于炎症在肿瘤进展中的作用的信息的质量和复杂性。Flow的核心工作人员在今年前三个季度培训了7名调查人员，让他们自己进行样本测试。训练有素的调查人员每天都在使用这些仪器，通常是在深夜和周末。这使得Flow核心工作人员有更多的时间在今年前三个季度对大约537个样本进行分类，并分析大约6700个样本。如果调查人员没有获取数据并分析自己的样本，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，至少发表了23篇论文，使用的是Flow Core获得的数据。在过去的一年里，从36名首席研究人员的实验室为78名科学家生成了数据(或对样本进行了分类)。虽然CCR-Frederick流式细胞术核心是CIP的一部分，但该项目的研究人员约占该设施使用的49%。未来的目标包括跟上流动实验室的尖端技术，将经过培训的研究人员池扩大到弗雷德里克的所有CCR，以运行和分析自己的样品，从而扩大使用核心仪器和专业知识产生的科学的用途、生产率和质量。这可以通过在LSRII Fortessa上增加一台405激光器，并用小型2激光6色仪器取代两台BD FACScans来实现。

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.