NEW APPROACHES TO FLOW SORTING OF CHROMOSOMES

染色体流式分选的新方法

• 批准号: 3508329
• 负责人: HOWARD M. SHAPIRO
• 金额: $ 10.42万
• 依托单位: HOWARD M. SHAPIRO, MD, P.C.
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-17 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3508329
• 关键词: artificial chromosomes; bioengineering /biomedical engineering; chromosomes; colorimetry; flow cytometry; fluorescent dye /probe; karyotype; method development; nucleobase; optics; ultraviolet spectrometry

Purification of individual chromosomes by flow sorting is often a first step in defining the structure of normal and abnormal human genes. DNA amplification techniques have greatly reduced the amount of sorted material required, making it feasible to develop smaller, lower-cost sorters. The performance of a dual-beam instrument using a single air- cooled helium-cadmium (He-Cd) laser as a source of both ultraviolet (UV) and blue excitation light for flow Cytometry of chromosomes stained with DAPI/olivomycin or Hoechst 33258/chromomycin A3 will be evaluated to determine whether the system can resolve the human karyotype as well as can existing commercial apparatus. In addition, several A-T and G-C preferring DNA fluorochromes not previously applied to chromosome analysis and potentially usable in a large number of existing flow sorters with single-beam 488 or 515 nm excitation will be examined as alternatives to stains presently used for bivariate flow karyotyping.

通过流式分选纯化单个染色体通常是第一步 定义正常和异常人类基因结构的一步。脱氧核糖核酸 扩增技术大大减少了排序量 所需材料，使得开发更小、成本更低的产品成为可能 分拣机。使用单个空气的双光束仪器的性能 冷却氦镉 (He-Cd) 激光器作为紫外线 (UV) 光源 和蓝色激发光，用于流式细胞仪染色的染色体 DAPI/橄榄霉素或 Hoechst 33258/铬霉素 A3 将被评估为 确定系统是否可以解析人类核型以及 可以使用现有的商业设备。另外，还有几个A-T和G-C 更喜欢之前未应用于染色体分析的 DNA 荧光染料 并有可能用于大量现有的流式分选机 将检查单光束 488 或 515 nm 激发作为替代方案 目前用于双变量流核型分析的染色剂。