Q：共染之后TUNEL 的信号就不在核内了，感觉都弥散了。TUNEL 染色就按照说明书来的，DAPI 之前孵育另外的一抗？

A：可能是后期洗涤次数过多，建议减少洗涤次数，或者洗涤动作轻柔一些。

Q：TUNEL 可以和 DAPI 一起染色细胞吗?

A：可以。

Q：Annexin V 和 JC-1、Tunel 细胞凋亡检测的区别？

A： Annexin V 是检测细胞早期凋亡的试剂，JC-1 是检测细胞中期凋亡的试剂、Tunel 是检测细胞晚期凋亡的试剂。

Q：Annexin V 和JC-1、Tunel 细胞凋亡检测的可以应用到植物或是细菌（原核生物） 吗？

A：可以，但是需要制备原生质体，因为植物细胞或是细菌（原核生物）含有细胞壁，具体的染液使用剂量只需浸没细胞即可，染色时间对于不同细胞有一定的不同。

Q：Tunel 细胞凋亡检测，细胞爬片好凋亡处理后需要在固定通透吗？

A：需要通透，因为 TdT 酶需要经过通透的细胞才能进入细胞内，而 Annexin V 和 JC-1 是不能进行染色固定的。

Q：Tunel 细胞凋亡检测时，贴壁细胞必须要先消化下来再染色吗？

A：不需要，对于贴壁细胞，要先用PBS 洗 2-3 次，然后直接用多聚甲醛固定细胞， 通透处理，染色观察。

Q：固定时间可以增加吗？

A： 4℃放置25 min左右，选择4%多聚甲醛作固定液，乙醇、甲醇、酸性固定液，会导致标记效率低；固定时间不宜过长，过长导致交联程度过高，进而降低标记效率。

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