牛带绦虫染色体标本制作的实验研究

Laboratory study of the beef tapeworm chromosome specimen handling

目的探讨绦虫染色体标本制作的影响因素。方法采用RPMI-1640培养液加秋水仙素体外培养—低渗处理—空气干燥技术,观察1640培养液不同液量、培养时间、秋水仙素不同剂量及不同的低渗液在标本制作过程中的效应。结果牛带绦虫成节生殖细胞有丝分裂指数在培养4h最高;培养液量以每节片6ml的1640培养液最好;秋水仙素剂量以0.1mg/ml为适宜,有良好的促进有丝分裂的作用;低渗液处理以0.075M氯化钾为佳,染色体清晰、分散程度好。结论以0.1mg/ml秋水仙素的1640培养液(6ml每节片)在37℃连续培养牛带绦虫生殖细胞4h,然后用0.075M氯化钾低渗空气干燥技术制作的染色体标本,效果最佳。

Objective To investigate the fa ctors affecting beef tapeworm chromosome specimen handling. Methods A hypoto nic management and air-d rying technique was used for beef tapeworm chromosome specimen handling with RPM I-1640 culture solution combined with colchicines in vitro. The outcomes were re corded during specimen handling with different 1640 culture solution, culture ti me, different dosage of colchicines and different hypotonic solution. Results The highest mitotic index of germ cells in progl ottids appeared at four hours after culture; the best solution were 1640 culture solution (6ml in every proglottid); the adaptable dosage of colchicines was 0.1mg/ml which can help th e mitosis; the best hypotonic management was with 0.075M potassiumichloride whic h can show clear chromosome and good dispersity. Conclusion In b eef tapeworm chromosome specimen handling, the best results will be obtained by which the beef tapeworm germ cells were cultured in vitro with 1640 culture solu tion (6ml in every proglottid) in combination with colchicines 0.1 mg/ml for 4 h ours at 37℃ continuously, then a hypotonic air-drying technique with which 0.07 5M potassiumichloride was also used.