使用说明：
1．贴壁细胞
  A.取洁净盖玻片在70%乙醇中浸泡5分钟或更长时间，无菌超净台内吹干或用无菌的PBS或0.9%NaCl等溶
    液洗涤三遍，再用细胞培养液洗涤一遍。将盖玻片置于六孔板内，种入细胞培养过夜，使约为50%-
    80%满。
  B.刺激细胞发生凋亡后，吸尽培养液，加入0.5ml 固定液，固定10分钟或更长时间(可4℃过夜)。
  C.去固定液，用PBS或0.9%NaCl 洗两遍，每次3分钟，吸尽液体。洗涤时宜用摇床，或手动晃动。
  D.加入0.5ml Hoechst 33258染色液，染色5分钟。也宜用摇床，或手动晃动数次。
  E.去染色液，用PBS或0.9%NaCl 洗两遍，每次3分钟，吸尽液体。洗涤时宜用摇床，或手动晃动。
  F.滴一滴抗荧光淬灭封片液于载玻片上，盖上贴有细胞的盖玻片，让细胞接触封片液，尽量避免气
    泡。
  G.荧光显微镜可检测到呈蓝色的细胞核。激发波长350nm左右，发射波长460nm左右，图谱请参考下
    图。
2.悬浮细胞
  A.离心收集细胞样品于1.5ml离心管内，加入0.5ml 固定液，缓缓悬起细胞，固定10分钟或更长时
    间(可4℃过夜)。
  B.离心去固定液，用PBS或0.9%NaCl 洗两遍，每次3分钟。洗涤期间手动晃动数次。
  C.离心后吸去大部分液体保留约50μl液体，再缓缓悬起细胞，滴加至载玻片上，尽量使细胞分布均
    匀。
  D.稍晾干，使细胞贴在载玻片上不易随液体流动。
  E.均匀滴上0.5ml Hoechst 33258染色液，染色5分钟。用吸水纸从边缘吸去液体，微晾干。
  F.去染色液，用PBS或0.9%NaCl 洗两遍，每次3分钟，吸尽液体。洗涤时宜用摇床，或手动晃动。
  G.滴一滴抗荧光淬灭封片液于载玻片上，盖上一洁净的盖玻片，尽量避免气泡。
  H.荧光显微镜可检测到呈蓝色的细胞核。激发波长350nm左右，发射波长460nm左右，图谱请参考下
    图。
3.组织切片
  A.常规包埋切片后，根据切片的不同类型，处理至可以用于免疫组化染色。
  B.PBS或0.9%NaCl 洗两遍，每次3分钟，吸尽液体。洗涤时宜用摇床，或手动晃动。可在六孔板中
    操作。
  C.加入0.5ml Hoechst 33258染色液，染色5分钟。也宜用摇床，或手动晃动数次。
  D.去染色液，用PBS或0.9%NaCl 洗两遍，每次3分钟，吸尽液体。洗涤时宜用摇床，或手动晃动。
  E.小心将切片置于载玻片上，滴一滴抗淬灭封片液，盖上一洁净的盖玻片，尽量避免气泡。
  F.荧光显微镜可检测到呈蓝色的细胞核。激发波长350nm左右，发射波长460nm左右，图谱请参考下
    图。
              
           Hoechst 33258的吸收光谱和发射光谱，左侧峰为吸收光谱，右侧峰为发射光谱。

     细胞凋亡坏死

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