隧道纳米管的诱导与观察综合性实验设计与实践

Design and practice of the comprehensive experiment of induction and detection of tunneling nanotubes

  • 摘要: 将细胞生物学前沿理论和研究技术及时转化到实验教学中,是提高本科实验教学水平和人才培养质量的重要途径之一.本实验以隧道纳米管(tunneling nanotubes,TNT)知识点为基础设计了“隧道纳米管的诱导与观察”综合性、探究性实验教学内容,包括血清饥饿模型的建立及隧道纳米管诱导、细胞微丝骨架及细胞核染色观察隧道纳米管结构、流式细胞术检测细胞周期3部分.首先针对不同类型的细胞探究合适血清质量分数建立血清饥饿模型,接着采用细胞爬片的方式进行细胞荧光染色,使用荧光显微镜观察桥状TNT结构,最后利用流式细胞术检测处于饥饿状态的细胞周期G1细胞比例的变化情况.教学实践表明,该实验有助于培养学生动手能力和科研思维,激发学生探索兴趣.

     

    Abstract: Induction and detection of tunneling nanotubes: comprehensive experimental design and practice. One important way to improve quality of undergraduate practical class is to transform cutting-edge theories and research techniques of cell biology into practical experiments. Knowledge of tunneling nanotubes (TNT) is used to design a comprehensive and exploratory experiment, including establishment of serum-starvation model and tunneling nanotubes induction, observation of tunneling nanotubes structure by staining of cell microfilaments and nuclei, and detection of cell cycle by flow cytometry. Students would first establish serum-starvation model by decreasing serum concentration for different cell types, then use cell crawler for fluorescence staining, and observe the bridge-like TNT structure under a fluorescence microscope. Flow cytometry is used to detect changes of the proportion of G1 cells in the cell cycle in starvation. These experiments facilitate student hands-on experience and stimulate their interest in exploration.

     

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