生物学杂志

生物学杂志 ›› 2023, Vol. 40 ›› Issue (2): 119-.doi: 10.3969/j.issn.2095-1736.2023.02.119

• 教学研究 • 上一篇    下一篇

Annexin V-FITC/PI法检测细胞凋亡实验

魏　远， 王宏刚， 陈成彬， 王春国， 宋文芹， 李登文， 赵立青   

  1. 南开大学 生物国家级实验教学示范中心， 天津 300071
  • 出版日期:2023-04-18 发布日期:2023-04-18
  • 通讯作者: 陈成彬，博士，副教授，研究方向为分子细胞遗传学，E-mail: chencb@nankai.edu.cn
  • 作者简介:魏远，硕士，实验师，从事细胞生物学实验和遗传学实验教学，E-mail: weiyuan@nankai.edu.cn
  • 基金资助:
    教育部2021年度基础学科拔尖学生培养计划2.0研究课题项目（20212017）； 南开大学2020年天津市高等学校本科教学质量与教学改革研究计划项目（B201005506）； 南开大学2021年本科教育教学改革项目（NKJG2021124）； 江西省自然科学基金资助项目(20202BABL213044)

Annexin V-FITC/PI detection of apoptosis experiment

WEI Yuan, WANG Honggang, CHEN Chengbin, WANG Chunguo, SONG Wenqin,LI Dengwen, ZHAO Liqing   

  1. National Demonstration Center for Experimental Biology Education, Nankai University, Tianjin 300071, China
  • Online:2023-04-18 Published:2023-04-18

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摘要： 课程组面向本科生开设了利用Annexin V-FITC/PI法检测细胞凋亡的实验项目。该实验加入化疗药物依托泊苷（简称VP-16）诱导Jurkat细胞发生凋亡，使用Annexin V-FITC/PI细胞凋亡检测试剂盒检测细胞凋亡情况，在荧光显微镜下拍照计数。实验结果表明：紫外光激发下，早期凋亡细胞呈现绿色荧光，而晚期凋亡细胞和坏死细胞呈现红色荧光；对细胞计数可知，对照组各时期细胞数符合正常范围，经药物处理后，各时期细胞凋亡率随药物浓度升高而升高，其中早期和晚期凋亡细胞占比最高；在高浓度药物处理后，细胞被大幅度杀伤，仅剩余几个早期凋亡细胞。实验将Annexin V-FITC/PI试剂盒与荧光显微镜结合使用，突破了教学经费、规模和仪器等条件局限，实验成本相对较低，适于大班教学，实验结果直观便捷，能够让学生深入、系统地掌握细胞凋亡的实验原理和检测方法，符合本科生实验项目的综合性高、设计性强和研究性佳的特点，切实提升细胞生物学实验课程的质量和水平。

关键词: 细胞凋亡, 实验教学, Annexin V-FITC/PI检测法, 荧光显微镜, 药物诱导

Abstract: An experimental course was set up to show how to detect apoptosis with Annexin V-FITC/PI to undergraduate students. Firstly, Etoposide (VP-16) was added to the Jurkat cells to induce the apoptosis, and then the apoptosis was detected by Annexin V-FITC/PI Apoptosis Detection Kit and photographed by fluorescence microscopy. The results showed that viable apoptotic cells showed green fluorescence under UV excitation, while late apoptotic and non-viable non-apoptoic cell showed red fluorescence. The number of cells was normal which had been counted in the control group. The apoptosis rate of different stages of cells increased to drug concentration increased after treated with drugs. The viable apoptotic cells and late apoptotic cells accounted for the highest proportion. While the cells were killed substantially, few early apoptotic cells remained when treated with high drug concentration. This experiment combined the Annexin V-FITC/PI kit with fluorescence microscopy, breaking the limitations of experimental conditions such as teaching funds, scale and instruments. Because this method costs relatively low, it is very suitable for large classes. On the other hand, the results are intuitive, so it is convenient for the students to understand the apoptosis principles and master the assay protocols. It meets the teaching requirements for undergraduate students teaching work, featured by high comprehensive, well-designed and researched. So that this apoptosis assay method effectively improves the level and quality of cell biology experimental teaching work.

Key words: apoptosis, experimental teaching, Annexin V-FITC/PI test, fluorescence microscope, drug induction

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