尿液样品处理

  1. 在室温 (15 - 30ºC)下,将尿液在 50 mL 离心管中以 600 g 离心 10 分钟。
  2. 去除上清,直至剩余的细胞沉淀物仅剩约 1 - 2 mL,注意不要干扰沉淀。
  3. 将沉淀重悬在剩余的 1 - 2 mL 上清液中,并将内容物转移到 15 mL 锥形离心管中。用 10 mL 1X PBS 冲洗 50 mL 试管,并将内容物转移到 15 mL 离心管中。
  4. 在室温下将样本以 600 g 离心 10 分钟。
  5. 去除上清,直至剩余的细胞沉淀物仅剩约 0.5 mL。
  6. 将沉淀重悬在剩余的 0.5 mL 上清液中。先缓慢滴加 1 - 5 mL 新鲜固定剂(3:1,甲醇:乙酸),先逐滴加入,而后反复振荡。
  7. 让固定的标本在 -20°C 下放置至少 30 分钟
  8. 在室温下将样本以 600 g 离心 5 分钟。小心去除上清。
  9. 将沉淀重悬于 1 - 5 mL 固定液中进行洗涤。
  10. 在室温下将样本以 600 g 离心 5 分钟。将步骤 8 和 9 重复两次。
  11. 将固定液中的细胞悬液离心后:
    • 如果细胞沉淀非常小且几乎看不见,小心除去尽可能多的固定剂,剩余约 100 µL 溶液。 
    • 如果细胞沉淀容易观察到,尽可能多地除去固定剂,并向细胞沉淀中加入 0.5 - 1 mL 新鲜固定剂。
  12. 立即根据玻片预处理方案继续进行后续步骤。继而使用 FISH 预处理试剂盒。 

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