小鼠骨髓细胞有丝分裂染色体标本制备方法的优化
|
摘要
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.
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.
引文
[1] 丁鸿,邱东萍,陈少雄.植物染色体标本的制备和染色体核型分析研究进展[J].南方农业学报,2012,43(12):1958-1962.
[2] 邓芳轶,杨玲.大蒜根尖有丝分裂染色体标本的制作[J].云南民族大学学报(自然科学版),2008,17(1):51-54.
[3] 王强.家蚕染色体的核型模式及Bmrbp1的FISH研究[D].重庆:西南大学,2006.
[4] 卢洪武,黎杰强,邓优美.小鼠骨髓细胞染色体G 带制片技术的探讨[J].实验室研究与探索,2014,33(1):46-51.
[5] 路玫,王佳丽,曹大明,等.针灸对环磷酰胺模型荷瘤小鼠骨髓细胞DNA修复相关蛋白动态调节的研究[J].中华中医药杂志,2016,31(8):3230-3233.
[6] 边高鹏,武有祯,焦海华.水胺硫磷致小鼠骨髓细胞DNA 损伤和氧化损伤[J].生态学杂志,2015,34(5):1413-1419.
[7] 闫玉军,徐文清,沈秀,等.银耳孢糖组分对137Cs γ 射线照射小鼠骨髓细胞染色体畸变及微核率的影响[J].中国辐射卫生,2014,23(1):54-56.
[8] 徐林.人类疾病的动物模型[J].动物学研究,2011,32(1):1-3.
[9] 薛丽香,张凤珠,孙瑞娟,等.我国疾病动物模型的研究现状和展望[J].中国科学:生命科学,2014,44(9):851-860.
[10] 刘恩岐,北岛修司,森本正敏.实验小鼠在人类癌症研究中的应用及其进展[J].癌症,2005,24(2):249-254.
[11] 刘涌涛,马全祥,刘慧娟.小鼠骨髓细胞染色体标本制备中的失误与对策[J].生物学通报,2003,38(6):53-54.
[12] 刘庆军,孙永君.小鼠骨髓细胞染色体标本制备方法的改进[J].山东轻工业学院学报,2012,26(4):64-65.
[13] 崔向清,马鑫,李丽,等.增加中期细胞数的染色体标本制备方法研究[J].中国现代医药杂志,2014,16(7):8-9.
[14] 谭军,曹向宇,蔡磊明.鹌鹑骨髓细胞染色体制片新方法[J].农药,2005,44(12):544-545,548.
[15] 姚应瑞.小白鼠骨髓细胞染色体制片技术[J].西安交通大学学报(医学版),1988(2):125.
[16] 黄燕,赵小平,余红,等.动物骨髓细胞染色体标本制备失败的原因分析[J].生物学通报,2006,41(1):52-53.
[17] 赵淑娟,庞有志,白俊艳,等.禽类骨髓细胞染色体标本制备实验方法的改进[J].实验动物科学,2010,27(2):17-20.
[18] 李碧春,徐银学.动物遗传学实验教程[M].3版.北京:中国农业大学出版社,2012.
[19] 王彬,孙晓玲,刘丽波.雄性小鼠生殖细胞染色体制备方法的研究[J].中国公共卫生,2004,20(3):259.
[20 ] 刘俊,李栋,张亚龙,等.小鼠骨髓染色体G显带最佳制备方法的探究[J].畜牧与饲料科学,2012,33(5/6):21-22.
[21] 朱娟娟,赵斌,刘莉茵,等.小鼠骨髓细胞染色体制备实验方法的改进[J].安徽医药,2007,11(8):768.
[22] 李芳,王文青,尹敬如,等.三项条件改变制备小鼠骨髓细胞染色体的分析[J].青海医学院学报,1990(2):153-155.
[2] 邓芳轶,杨玲.大蒜根尖有丝分裂染色体标本的制作[J].云南民族大学学报(自然科学版),2008,17(1):51-54.
[3] 王强.家蚕染色体的核型模式及Bmrbp1的FISH研究[D].重庆:西南大学,2006.
[4] 卢洪武,黎杰强,邓优美.小鼠骨髓细胞染色体G 带制片技术的探讨[J].实验室研究与探索,2014,33(1):46-51.
[5] 路玫,王佳丽,曹大明,等.针灸对环磷酰胺模型荷瘤小鼠骨髓细胞DNA修复相关蛋白动态调节的研究[J].中华中医药杂志,2016,31(8):3230-3233.
[6] 边高鹏,武有祯,焦海华.水胺硫磷致小鼠骨髓细胞DNA 损伤和氧化损伤[J].生态学杂志,2015,34(5):1413-1419.
[7] 闫玉军,徐文清,沈秀,等.银耳孢糖组分对137Cs γ 射线照射小鼠骨髓细胞染色体畸变及微核率的影响[J].中国辐射卫生,2014,23(1):54-56.
[8] 徐林.人类疾病的动物模型[J].动物学研究,2011,32(1):1-3.
[9] 薛丽香,张凤珠,孙瑞娟,等.我国疾病动物模型的研究现状和展望[J].中国科学:生命科学,2014,44(9):851-860.
[10] 刘恩岐,北岛修司,森本正敏.实验小鼠在人类癌症研究中的应用及其进展[J].癌症,2005,24(2):249-254.
[11] 刘涌涛,马全祥,刘慧娟.小鼠骨髓细胞染色体标本制备中的失误与对策[J].生物学通报,2003,38(6):53-54.
[12] 刘庆军,孙永君.小鼠骨髓细胞染色体标本制备方法的改进[J].山东轻工业学院学报,2012,26(4):64-65.
[13] 崔向清,马鑫,李丽,等.增加中期细胞数的染色体标本制备方法研究[J].中国现代医药杂志,2014,16(7):8-9.
[14] 谭军,曹向宇,蔡磊明.鹌鹑骨髓细胞染色体制片新方法[J].农药,2005,44(12):544-545,548.
[15] 姚应瑞.小白鼠骨髓细胞染色体制片技术[J].西安交通大学学报(医学版),1988(2):125.
[16] 黄燕,赵小平,余红,等.动物骨髓细胞染色体标本制备失败的原因分析[J].生物学通报,2006,41(1):52-53.
[17] 赵淑娟,庞有志,白俊艳,等.禽类骨髓细胞染色体标本制备实验方法的改进[J].实验动物科学,2010,27(2):17-20.
[18] 李碧春,徐银学.动物遗传学实验教程[M].3版.北京:中国农业大学出版社,2012.
[19] 王彬,孙晓玲,刘丽波.雄性小鼠生殖细胞染色体制备方法的研究[J].中国公共卫生,2004,20(3):259.
[20 ] 刘俊,李栋,张亚龙,等.小鼠骨髓染色体G显带最佳制备方法的探究[J].畜牧与饲料科学,2012,33(5/6):21-22.
[21] 朱娟娟,赵斌,刘莉茵,等.小鼠骨髓细胞染色体制备实验方法的改进[J].安徽医药,2007,11(8):768.
[22] 李芳,王文青,尹敬如,等.三项条件改变制备小鼠骨髓细胞染色体的分析[J].青海医学院学报,1990(2):153-155.