Q：染色结果可以使用多功能酶标仪观察吗？

A：可以，建议如果条件允许最好是使用荧光显微镜或是共聚焦显微镜 流式仪器。

Q：染色后多久进行？

A：建议染色后应立即进行观察，应当在 30min 内完成整个实验观察过程。

Q：染色工作液使用什么溶液进行配置？

A：如果试剂盒内提供有稀释 buffer，应使用该溶液配置，如果没有配备，可以使用 HBSS 无血清培养基 PBS 等合适的缓冲液进行配置。

Q：染色时添加多少体系的工作液进行染色？

A：如果是爬片观察，添加的剂量应是尽量浸没细胞为宜，如果采用流式观察使用剂量可以参考说明书建议的用量。

Q：40307 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：40307 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，在通透处理，染 色观察。

Q：可以通过那些仪器检测tunel 荧光信号？

A：荧光显微镜 共聚焦显微镜 流式细胞仪 酶标仪均可以检测tunel 荧光信号。

Q：Tunel 检测在实验操作上有哪些注意事项？

A：荧光基团长期暴露在普通光照下，会发生严重淬灭现象，因此在实验操作上应尽量避光操作和孵育。

Q：Tunel 检测除了多聚甲醛固定，可不可以使用甲醇固定？

A：使用乙醇或甲醇固定会导致标记的效率较低。酸性固定液，也容易导致假阳性的出现。建议采用新鲜配置的 4%中性多聚甲醛固定液。

Q：试剂盒的保质期多久？

A：对于探针类的试剂在规定的保存条件下保质期是一年左右。

Q：荧光显微镜检测时发现荧光较弱，怎么办？

A：对于荧光较弱的情况可以适当的增加孵育时间和工作液浓度，对于细胞也尽量选择状态较好的细胞进行染色观察。

Q：为什么我激发后荧光很快就猝灭了？

A：可能是使用浓度过高造成，可以再适当降低一些试剂浓度；可以在寻找视野时使用低能量的激光， 待找到合适视野时使用高能量激光激发观察；或者是使用抗猝灭剂。

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