CORE--CELL SORTING

核心——细胞分选

• 批准号: 7142508
• 负责人: Robert C Young
• 金额: $ 7.36万
• 依托单位: FOX CHASE CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-10 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7142508
• 关键词: biomedical facility; cell sorting; flow cytometry; immunofluorescence technique; neoplasm /cancer; nephelometry

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.

研究细胞分选和流式细胞术设施提供福克斯大通癌症中心 （FCCC）研究了通过多达14个独立测量分析细胞的能力，其中12个基于免疫荧光，两个基于光散射。该设施用于 36名经同行评审的受资助研究人员（比上次评审增加21名或71%） 在9个研究项目和调查人员在所有三个FCCC司。九十五 其中95%（95%）是由同行评审资助的研究人员使用的。流式细胞仪的主要应用包括细胞亚群的鉴别和分离（分选）（高达20，000个细胞/秒），罕见变异（1/10[7]）细胞系的分离，肿瘤或细胞系的表征，以及通过直接沉积到96孔板中的细胞系的单细胞克隆。除了提供更多的荧光检测器（从5个增加到12个）外，最近升级的分选机还提供高速4流分选和数字检测。具有紫外线功能的激光器可以测量钙通量和活细胞DNA含量。除了3激光12色细胞分选仪（FACS-VantageSE/DiVa）外，该设备还包括一台仅用于分析的8色LSR-II和一台三色FACScan。该设施为用户提供有关实验设计（包括试剂和适当荧光染料的可用性）、样品制备和数据解释的建议。与FCCC生物信息学小组合作开发的软件Flow-LIMS用于数据注释、组织和存档。分析是在研究人员的实验室计算机上使用高度互动的图形软件程序Flowjo进行的。自上次审查以来，小时使用量（目前估计超过理论容量的80%）增加了65%，预计在未来五年内将以类似的方式增加。公平使用该设施是由一个学院监督委员会保证。