小鼠骨髓细胞有丝分裂染色体标本制备方法的优化

Optimization of Preparation Methods for Mitotic Chromosome Specimen of Bone Barrow Cells in Mice

将201只小鼠随机分为3组,分别采用1次固定(60 min)、2次固定(30、25 min)和3次固定(30、25和15 min),其他操作步骤相同。结果表明,与1次固定相比,2次固定和3次固定视野中骨髓细胞数量(P<0.05)和分裂相细胞数量显著增多(P<0.05),制片成功率分别高7.1和7.9百分点,固定时间分别增加5和30 min。1次固定整体效果不佳,或很难找到明确形态的染色体,或染色体形态不清晰、分散;2次固定染色体分散效果良好,染色体数目和形态都十分清晰,染色体结构完整;3次固定中期分裂相多,染色体集中且分散适度,染色体清楚可见,分布整齐不重。综合考虑,小鼠骨髓细胞有丝分裂染色体标本制备的最佳固定次数为2次,适宜的固定时间为30、25 min。

A total of 201 mice were randomly divided into 3 groups, which were fixed one time (60 min), two times (30, 25 min) and three times (30, 25,15 min), and the other procedure were not different. The results showed that, compared with fixing once, the number of bone marrow cells and split phase cell in the vision field of fixing two times and fixing three times significantly increased( P <0.05),the production success rate in the methods of fixing two times and fixing three times increased 7.1 and 7.9 percentage point respectively, and the fixing time in the methods of fixing two times and fixing three times increased 5 and 30 min respectively. As for fixing once,the overall effect was not good, it was difficult to find definite shape of chromosome, or the chromosome shape was dispersed. As for fixing two times,the number and shape of chromosomes were clear, the dispersion effect was good and the structure of chromosomes was complete. As fixing three times,many cells were in metaphase,the chromosome were concentrated and dispersed moderately,and the chromosomes were clear. Considering comprehensively,the best fixing times for the preparation of mitotic chromosomes in mice bone marrow cells was two times, and the suitable fixation time was 30 and 25 min, respectively.