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Jingdong book

流式细胞术的基础和临床应用

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流式细胞术的基础和临床应用

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流式细胞术(flow cytometry,FCM)是利用流式细胞仪对单细胞或其他生物粒子进行定量分析和分选。流式细胞术综合运用电子、激光、计算机以及流体力学等方法,可在极短的时间内高速分析上万个细胞,同时从一个细胞中测得多个参数。与传统的荧光显微镜检查相比,具有速度快、精度高、准确性好等优点,成为当代最先进的细胞定量分析技术。更值得注意的是,流式细胞术可以从单细胞水平检测细胞的免疫表型特征及生物学功能的变化。近几年来,国内外对流式细胞术的发展做了不少的研究工作,目前人们逐步对流式细胞术所需要的样品的制备、标记方法及应用范围等进行了深入的研究。
Catalogue

第一章 总论
第一节 流式细胞术与流式细胞仪概述
一、流式细胞术概述
二、流式细胞仪
(一)基本结构
(二)工作原理
(三)分析型流式细胞仪介绍
(四)分选型流式细胞仪介绍
第二节 荧光染料的选择与常用的染色技术
一、荧光染料的特性及其应用
二、常用的染色技术
(一)细胞免疫荧光染色技术
(二)核酸荧光染色技术
(三)荧光微球捕获染色技术
第三节 结果分析与流式图
一、流式通道
二、流式直方图
三、流式散点图
四、流式等高线图
第四节 流式细胞术的基本操作与技巧
一、标本的采集、运送与储存
二、标本的处理
(一)独立细胞样品制备
(二)免疫器官样品制备
(三)实体脏器样品制备
三、荧光素偶联抗体及其标记方法
(一)荧光素与荧光素偶联抗体
(二)样品封闭
(三)荧光素偶联抗体标记
四、光电倍增管电压设定
五、对照的设置
(一)阴性对照的设置
(二)FMO对照
(三)阳性对照的设置
六、补偿调节
(一)补偿调节的原理
(二)调节补偿的具体方法
(三)三色和四色分析补偿调节方法
(四)影响补偿大小的因素
七、阈值设定
(一)减少样品中死细胞比例的方法
(二)流式分析时区分死细胞和活细胞的方法
八、流式分选模式选择
(一)纯度模式
(二)得率模式(富集模式)
(三)单细胞模式
九、上样速度控制
(一)流式分析速度控制
(二)流式分选速度控制
十、分选设门基本原则
十一、流式分选基本步骤
第五节 方法学特点
一、单细胞分析
二、定量分析
三、快速性
四、准确性
五、特异性
六、可重复性
七、多参数分析
第六节 常见影响因素
一、温度
二、pH
三、荧光染料浓度
四、杂质
五、细胞固定剂
六、溶液黏度
七、其他因素
……
第二章 流式分析术及流式分选术的应用
第三章 流式细胞术的临床应用

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