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流式细胞仪检测 T 细胞亚型

流式细胞仪检测 T 细胞亚型. 实验目的. 掌握流式细胞仪的原理、方法和流式结果分析。 了解流式细胞仪的应用。. Flow Cytometry. Flow Tip. FL Detector. Light Scatter Detector. Laser. Flow Cytometry. Flow Cytometry. One Parameter Histogram. Two Parameter Histogram. Unstained cells. Green Fluorescence Intensity. FITC-labeled cells.

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流式细胞仪检测 T 细胞亚型

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  1. 流式细胞仪检测T细胞亚型

  2. 实验目的 • 掌握流式细胞仪的原理、方法和流式结果分析。 • 了解流式细胞仪的应用。

  3. Flow Cytometry Flow Tip FL Detector Light Scatter Detector Laser

  4. Flow Cytometry

  5. Flow Cytometry One Parameter Histogram Two Parameter Histogram Unstained cells Green Fluorescence Intensity FITC-labeled cells Number of Cells Red Fluorescence Intensity Green Fluorescence Intensity

  6. 实验材料 • 外周血单个核细胞 • 洗涤液 • 固定液

  7. 实验步骤 制备活性高的细胞悬液　　　　　　　　　　　↓ 用10％FCS RPMI1640调整细胞浓度为5×10６～１×10７／ml　　　　　　　　　　　↓ 取40μl细胞悬液加入50μl工作浓度的荧光标记抗体，充分振摇　　　　　　　　　　 ↓　4℃ 30min　　用洗涤液洗涤2次，每次加液2ml左,1000rpm×5min　　　　　　　　　　　↓ 加适量固定液(如为FCM制备标本，一般加入1ml固定液，如制片后在荧光显微镜下观察，视细胞浓度加入100～500μl固定液)　　　　　　　　　　 ↓ FCM检测或制片后荧光显微镜下观察(标本在试管中可保存5～7天)

  8. 注意事项 • 整个操作在4℃下进行，洗涤液中加有比常规防腐剂量高10倍的NaN３，上述实验条件是防止一抗结合细胞膜抗原后发生交联、脱落。 • 洗涤要充分，以避免出现假阴性。 • 细胞活性要好，否则易发生非特异性荧光染色。

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