CORE--CELL SORTING

研究细胞分选和流式细胞术设施为福克斯·蔡斯癌症中心提供 (Fccc)调查人员通过多达14种独立测量来分析细胞的能力，其中12种是基于免疫荧光，两种是基于光散射。此设施由以下人员使用 36名同行审查资助的调查员(比上次审查的21人或71%有所增加) 在9个研究项目中，以及由联合国气候变化框架公约所有三个部门的调查人员。九十五 其中95%的使用是由同行评议资助的调查人员完成的。流式细胞仪的主要应用包括识别和分离(分选)细胞亚群(高达20,000个/秒)，分离罕见的变异细胞系(1/10[7])，肿瘤或细胞系的特征，以及通过直接沉积到96孔板中对细胞系进行单细胞克隆。除了提供更多的荧光检测器(从5个增加到12个)外，最近升级的分拣机还提供高速4流分选和数字检测。具有紫外线功能的激光可以测量钙离子通量和活细胞DNA含量。除了3激光12色细胞分选机(FACS-VantageSE/DIVA)外，该设施还包括一台仅供分析的8色LSR-II和一台三色FACScan。该设施向用户提供关于实验设计(包括试剂和适当的荧光试剂的可用性)、样品制备和数据解释的建议。与fccc生物信息学小组共同开发的软件Flow-LIMS用于数据注释、组织和存档。分析是在调查人员的实验室计算机上使用高度交互的图形软件程序FlowJo进行的。自上次审查以来，小时使用量(目前估计超过理论能力的80%)增加了65%，预计未来五年也将同样增加。学院监督委员会保证公平使用该设施。

The Research Cell Sorting and Flow Cytometry Facility offers Fox Chase Cancer Center (FCCC) investigators the capability of analyzing cells by up to 14 independent measurements, 12 of which are based on immunofluorescence and two on light scattering. This Facility is used by 36 peer-reviewed funded investigators (an increase from 21 or 71% from the previous review) across 9 Research Programs and by investigators within all three FCCC Divisions. Ninety-five percent (95%) of this usage is by peer-reviewed funded investigators. Major applications of the flow cytometer include discrimination and isolation (sorting) of cell subpopulations (at up to 20,000 cells/sec), isolation of rare variant (1/10[7]) cell lines, characterization of tumors or cell lines, and single cell cloning of cell lines by direct deposition into 96-well plates. In addition to providing many more fluorescence detectors (increase from 5 to 12), the recently upgraded sorter provides high-speed 4-stream sorting and digital detection. A UV-capable laser enables measurement of calcium flux and viable cell DNA content. In addition to the 3-laser 12-color cell sorter (FACS-VantageSE/DiVa), the Facility also includes an analysis-only 8-color LSR-II and a three-color FACScan. The Facility aids users with advice on experimental design (including availability of reagents and appropriate fluorochromes), sample preparation, and data interpretation. Software developed with the FCCC Bioinformatics group, the Flow-LIMS, is used for data annotation, organization, and archiving. Analysis is done on investigator's laboratory computers using a highly interactive, graphical software program, FlowJo. Hour usage (currently estimated at over 80% of theoretical capacity) has increased 65% since the last review and is projected to increase similarly over the next five years. Equitable access to the Facility is assured by a Faculty Oversight Committee.