24小时热门版块排行榜

返回列表

【奖励】本帖被评价141次，作者shenyingbin增加金币 111.4 个

shenyingbin

铁杆木虫 (著名写手)

[资源] 流式细胞仪使用手册（中文版）

仪器类型：BD FACSCalibur
目录：
第一章流式细胞仪简介...............................................................................................1
1.1 流式细胞术发展史...........................................................................................1
1.2 流式细胞仪构造和工作原理...........................................................................2
1.2.1 概述.........................................................................................................2
1.2.2 流式细胞仪构造.....................................................................................2
1.3 流式细胞仪的主要技术指标.........................................................................10
第二章 FACSCalibur 的日常操作...............................................................................12
2.1 FACSCalibur系统...........................................................................................12
2.2 FACSCalibur开机程序...................................................................................15
2.3 FACSCalibur关机程序...................................................................................16
2.4 FACSCalibur的维护与保养...........................................................................17
2.4.1 FACSCalibur的每月维护......................................................................17
2.4.2 FACSCalibur的定期维护......................................................................18
2.5 常见故障排除.................................................................................................20
第三章 FACSComp软件..............................................................................................22
3.1 FACSComp简介.............................................................................................22
3.1.1 FACSComp运行条件............................................................................22
3.1.2 FACSComp的文件类型........................................................................23
3.2 FACSComp的功能.........................................................................................24
3.2.1 光电倍增管（PMT）电压的调节.......................................................24
3.2.2 荧光补偿的调节...................................................................................24
3.2.3 灵敏度的测试.......................................................................................24
3.2.4 时间延迟的校准（对于双激光，配有FL4 PMT）............................25
3.2.5 HLA-B27的校准...................................................................................25
3.2.6 LeucoCOUNT........................................................................................25
3.2.7 优化.......................................................................................................25
3.3 FACSComp的运行.........................................................................................25
3.3.1 CaliBRITE Beads的准备.......................................................................25
3.3.2 软件环境的设置...................................................................................26
3.3.3 Beads的检测..........................................................................................26
3.3.4 Optimization..........................................................................................28
3.4 Troubleshooting...............................................................................................28
第四章 FACStation 数据管理系统..............................................................................30
4.1 FACStation 数据管理系统组成.....................................................................30
4.2 FACStation文件组成......................................................................................31
4.2.1 如何建立文件夹...................................................................................31
4.2.2 FACStation文件的类型.........................................................................31
4.2.3 如何管理文件.......................................................................................32
4.3 实用的Macintosh OS X功能........................................................................34
4.3.1 菜单栏...................................................................................................35
4.3.2 Dock.......................................................................................................37
4.3.3 查找窗口...............................................................................................37
4.3.4 键盘快捷键...........................................................................................38
4.4 如何设置模版.................................................................................................39
4.5 实验练习：如何将ModFit LT的别名添加到Dock菜单下...........................40
4.6 可选性存储装置.............................................................................................40
4.6.1 可选性存储装置的基本工作原则.......................................................40
4.6.2 光驱的维护...........................................................................................41
4.6.3 光盘的维护...........................................................................................41
第五章 CellQuest Pro软件...........................................................................................42
5.1 概述.................................................................................................................42
5.1.1 获取(Acquisition）................................................................................42
5.1.2 分析(Analysis).......................................................................................42
5.1.3 从获取到分析(Acquisition Analysis)..................................................42
5.2 工具栏.............................................................................................................42
5.3 CellQuest Pro 的文件.....................................................................................43
5.3.1 FCS数据文件........................................................................................43
5.3.2 实验文件...............................................................................................44
5.3.3 仪器条件文件.......................................................................................44
5.3.4 统计文件...............................................................................................44
5.4 CellQuest Pro 的仪器控制.............................................................................44
5.4.1 探测器...................................................................................................45
5.4.2 放大器...................................................................................................45
5.4.3 阈值......................................................................................................46
5.4.4 补偿......................................................................................................46
5.4.5 状态......................................................................................................47
5.5 CellQuest Pro 的视图.....................................................................................48
5.6 练习：3色或4色数据获取.............................................................................48
5.6.1 质控－运行FACSComp........................................................................48
5.6.2 优化.......................................................................................................50
5.6.3 调出储存的由FACSComp 生成的仪器条件.......................................52
5.6.4 调节FSC/SSC探测器（电压）及FSC阈值.........................................53
5.6.5 设置淋巴细胞Region...........................................................................53
5.6.6 调节FL1、FL2、FL3的探测器（电压）...........................................53
5.6.7 调节荧光补偿.......................................................................................55
5.7 实验练习：3色/4色预获取............................................................................55
5.7.1 设置Acquisition & Storage 窗口...........................................................56
5.7.2 设置Parameter Description....................................................................57
5.7.3 编辑Reagent Panel.................................................................................58
5.8 实验练习：数据获取.....................................................................................59
5.9 储存、恢复仪器设置.....................................................................................60
5.9.1 储存仪器设置.......................................................................................60
5.9.2 恢复仪器设置.......................................................................................60
5.10 储存实验模板文件.......................................................................................60
5.11 练习：数据分析...........................................................................................60
5.11.1 画Region..............................................................................................61
5.11.2 限定象限marker..................................................................................61
5.11.3 分析四色..............................................................................................63
5.11.4 批分析其余数据..................................................................................63
5.11.5 分析数据..............................................................................................64
5.11.6 将分析文件以模板形式保存..............................................................64
5.11.7 创建文具簿（stationery pad）...........................................................64
5.11.8 将统计改为电子表格..........................................................................65
5.12 Regions 和Gating（画门）..........................................................................65
5.12.1 设置Region..........................................................................................65
5.12.2 改变Region..........................................................................................65
5.12.3 用Region 统计来分析数据.................................................................66
5.12.4 门.........................................................................................................66
5.12.5 多色门.................................................................................................66
5.12.6 组合门.................................................................................................67
5.12.7 在直方图上使用组合门分析.............................................................68
第六章 FACSCalibur 分选...........................................................................................69
6.1 分选概述.........................................................................................................69
6.1.1 收集系统...............................................................................................69
6.1.2 分选模式...............................................................................................70
6.2 分选窗口.........................................................................................................71
6.2.1 Sort Setup...............................................................................................71
6.2.2 Sort Counters.........................................................................................72
6.3 准备收集管.....................................................................................................72
6.4 分选.................................................................................................................73
6.4.1 清洗分选管线.......................................................................................73
6.4.2 准备.......................................................................................................73
6.4.3 收集分选前数据...................................................................................74
6.4.4 分选条件设置.......................................................................................74
6.4.5 分选目的细胞.......................................................................................74
6.4.6 清洗分选管路.......................................................................................75
6.4.7 浓缩样本...............................................................................................76
6.5 检验分选纯度.................................................................................................76
6.6 分选问题讨论.................................................................................................77
6.7 FACSCalibur无菌分选...................................................................................79
6.8 分选细胞浓缩系统.........................................................................................80
6.8.1 系统简介................................................................................................80
6.8.2 细胞培养基嵌入物和滤膜的制备.......................................................81
6.8.3 确定气压参数........................................................................................82
6.8.4 如何用细胞浓缩系统进行分选............................................................82
6.8.5 分选浓缩细胞.......................................................................................84
6.8.6 从分选管路中回收细胞.......................................................................84
6.8.7 回收细胞做进一步分析.......................................................................85
6.8.8 清洗分选管路.......................................................................................85
6.8.9 清洗浓缩器...........................................................................................85
第七章 DNA分析.........................................................................................................87
7.1 概述.................................................................................................................87
7.2 细胞周期.........................................................................................................87
7.3 DNA检测的常用术语....................................................................................87
7.3.1 Coefficient of Variation（CV）：变异系数........................................87
7.3.2 DNA index（DI）：DNA指数............................................................88
7.3.3 Ploidy：倍体.........................................................................................88
7.4 影响流式DNA分析的因素............................................................................88
7.4.1 分辨率（CV）......................................................................................88
7.4.2 线性度...................................................................................................88
7.4.3 碎片.......................................................................................................88
7.4.4 细胞双粘体...........................................................................................89
7.5 DNA质量控制（DNA QC Particles）..........................................................89
7.5.1 样本制备...............................................................................................89
7.5.2 上机检测...............................................................................................89
7.5.3 常见错误排除.......................................................................................90
7.6 用CellQuest Pro软件获取DNA 数据.............................................................91
7.6.1 样本制备...............................................................................................91
7.6.2 用CellQuest Pro软件上机获取数据.....................................................92
7.6.3 质量控制...............................................................................................95
7.7 用ModFit 软件分析DNA数据.......................................................................95
7.7.1 运用ModFit 进行自动分析..................................................................96
7.7.2 运用ModFit 进行手动分析..................................................................96
7.7.3 运用ModFIT进行同步化分析..............................................................98
7.7.4 运用ModFIT 进行增殖分析.................................................................98
第八章 HLA-B27分析.................................................................................................99
8.1 检测HLA-B27的意义.....................................................................................99
8.2 HLA-B27的检测方法.....................................................................................99
8.2.1 传统方法...............................................................................................99
8.2.2 流式细胞术.........................................................................................100
8.2.3 试剂盒介绍.........................................................................................100
8.2.4 实验原理.............................................................................................100
8.2.5 样本的收集和准备.............................................................................101
8.2.6 流式检测.............................................................................................102
8.3 质量控制.......................................................................................................110
附录1：组织相容性抗原和疾病的关系........................................................... 111
附录2：强直性脊柱炎——一个常见但易被忽略的疾病................................112
第九章 MultiSET软件........................................................................................113
9.1 简介...............................................................................................................113
9.2 MultiSET运行条件.......................................................................................114
9.3 MultiSET文件类型.......................................................................................115
9.4 MultiSET软件内容.......................................................................................116
9.4.1 MultiSET界面功能分区.....................................................................116
9.4.2 MultiSET命令菜单.............................................................................116
9.4.3 MultiSET实验报告.............................................................................117
9.4.4 MultiSET运行程序.............................................................................117
9.5 利用Multiset软件的Tools和Preferences......................................................126
9.6 Control Panels (绝对计数质控)....................................................................128
9.7 MultiSET画门及Attractor方案.....................................................................128
9.8 注意事项及常见问题处理...........................................................................129

回复此楼

» 本帖附件资源列表

欢迎监督和反馈：小木虫仅提供交流平台，不对该内容负责。
本内容由用户自主发布，如果其内容涉及到知识产权问题，其责任在于用户本人，如对版权有异议，请联系邮箱：xiaomuchong@tal.com
附件 1 : BD_FACSCalibur中文培训手册.pdf

2015-05-13 09:04:59, 4.48 M

» 收录本帖的淘帖专辑推荐

蛋白表达纯化鉴定精华	科研之家（食品）	酿酒实验室	实验中的策略
待下资源	流式细胞	肿瘤药理学相关	流式细胞术
食品科学与工程