摘要
目的深入分析预固定在淋巴细胞染色体制备中的机制及影响因素,为染色体量化制备提供依据。方法针对预固定环节中甲醇与冰醋酸的体积配比、用量及作用时间进行试验,以染色体分裂指数及分裂相合格率为指标,研究预固定环节各因素对染色体制备的影响。结果预固定液中甲醇与冰醋酸的体积配比对分裂指数影响差异无统计学意义(F=1. 549,P> 0. 05),对分裂相合格率影响差异有统计学意义(F=8. 192,P <0. 05),预固定液中甲醇与冰醋酸体积比可控制在3∶1～1∶1。预固定液用量对分裂指数(F=5. 327,P <0. 05)和分裂相合格率(F=4. 989,P <0. 05)影响差异有统计学意义,将预固定液用量维持在0. 8～0. 9 ml,染色体分散良好,无重叠。预固定作用时间对分裂指数(F=0. 821,P> 0. 05)和分裂相合格率(F=0. 458,P> 0. 05)影响差异无统计学意义,推荐预固定处理时间为1 min。结论预固定液中甲醇与冰醋酸配比控制在3∶1～1∶1之间,预固定液用量控制在0. 8～0. 9 ml之间,预固定1 min,可获得理想的染色体分裂相,将预固定环节影响因素进行量化分析,为预固定实现规范操作打下基础。
Objective To deeply research the mechanism and influencial factors of prefixation in chromosome preparation,to make a reference for quantitative chromosome preparation. Methods Aiming at the volume ratio,dosage and duration of methanol and glacial acetic acid in the pre-fixation process,the effects of various factors of pre-fixation on chromosome preparation were studied by using the chromosomal splitting index and the rate of division of the split phase. Results The proportion of methanol and glacial acetic acid had no statistically significant difference on mitotic index( F = 1. 549,P > 0. 05),but on the qualified rate of metaphase chromosome spreading( F = 8. 192,P < 0. 05). The volume ratio of methanol to glacial acetic acid in the pre-fixed solution can be controlled from 3∶1 to 1∶1. The volume of prefixative had statistically significant difference on mitotic index( F = 5. 327,P < 0. 05) and qualified rate of metaphase chromosome spreading( F = 4. 989,P < 0. 05). The dosage of the pre-fixed solution was maintained at 0. 8 ~ 0. 9 ml,and the chromosomes were well dispersed without overlap. The factor of action time had no statistically significant difference on the mitotic index( F = 0. 821,P > 0. 05) and qualified rate of metaphase chromosome spreading( F = 0. 458,P > 0. 05). The recommended pre-fixation processing time was 1 min. Conclusion The ratio of methanol and glacial acetic acid in the pre-fixed solution is under the range about 3∶1 ~ 1∶1,and the amount of pre-fixed solution is controlled between 0. 8 and 0. 9 ml. Pre-fixed for 1 min to obtain the ideal chromosome splitting phase. The quantitative analysis of the influencing factors of the fixed link lays the foundation for the pre-fixed implementation of the standard operation.
作者
杨秋月
赵俊鹏
王玉君
蔡超
秦磊
YANG Qiuyue;ZHAO Junpeng;WANG Yujun;CAI Chao;QIN Lei(Tangshan City Center for Disease Control and Prevention,Hebei063000 ,China)
出处
《医学动物防制》
2019年第5期509-512,共4页
Journal of Medical Pest Control
基金
河北省2018年度医学科学研究重点课题计划(20181383)
关键词
染色体
预固定
中期分裂相
量化分析
Chromosome
Prefixation
Metaphase chromosome spreading
Quantitative analysis
