[膜联蛋白V-异硫氰酸荧光素/碘化丙啶与Hoechst33342/碘化丙啶双染色在流式细胞术检测细胞凋亡中的比较]

[Comparison between annexin V-FITC/PI and Hoechst33342/PI double stainings in the detection of apoptosis by flow cytometry].

作者信息

Zhang Wei, Liang Zhihui

机构信息

Department of Basic Medicine, Medical College, Wuhan University of Science and Technology, Wuhan 430065, China.

Cell Engineering Centre, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2014 Nov;30(11):1209-12.

PMID:25374090

Abstract

OBJECTIVE

To evaluate the merits and demerits of annexin V-FITC/PI and Hoechst33342/PI double stainings in the detection of apoptosis by flow cytometry.

METHODS

Hoechst33342/PI and annexin V-FITC/PI double stainings were performed to detect the apoptosis of thymocytes induced by DEX and K562 cells induced by H(2)O(2).

RESULTS

The anenxin V-FITC/PI double staining was able to detect the cell apoptosis in both DEX-induced thymocytes and H(2)O(2)-induced K562 cells as predicted, and the cell plotting was consistent with the principle of the method. Hoechest33342/PI double staining was able to detect cell apoptosis in the DEX-induced thymocytes, but this method showed a contradictory result in the early stage of H(2)O(2)-induced K562 cell apoptosis, and the cell plotting was in conflict with the principle of the method.

CONCLUSION

Compared with annexin V-FITC/PI double staining, the method of Hoechst33342/PI double staining in the detection of apoptosis by flow cytometry has some limitations.

摘要

目的

评价膜联蛋白V-异硫氰酸荧光素/碘化丙啶（Annexin V-FITC/PI）和双苯并咪唑33342/碘化丙啶（Hoechst33342/PI）双染色法在流式细胞术检测细胞凋亡中的优缺点。

方法

采用Hoechst33342/PI和Annexin V-FITC/PI双染色法检测地塞米松（DEX）诱导的胸腺细胞及过氧化氢（H₂O₂）诱导的K562细胞的凋亡情况。

结果

Annexin V-FITC/PI双染色法能够如预期检测到DEX诱导的胸腺细胞及H₂O₂诱导的K562细胞的凋亡，细胞散点图符合该方法原理。Hoechest33342/PI双染色法能够检测到DEX诱导的胸腺细胞凋亡，但该方法在H₂O₂诱导的K562细胞凋亡早期出现矛盾结果，细胞散点图与该方法原理不符。