A COLCHICINE, HYPOTONIC CITRATE, SQUASH SEQUENCE FOR MAMMALIAN CHROMOSOMES
A COLCHICINE, HYPOTONIC CITRATE, SQUASH SEQUENCE FOR MAMMALIAN CHROMOSOMES
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DOI：
10.3109/10520295609113814
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发表时间：
1956-01-01
期刊：
影响因子：
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通讯作者：
HAMERTON, JL
中科院分区：
文献类型：
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作者：
FORD, CE;HAMERTON, JL
该方法仅适用于小鼠骨髓，但适用于其他组织和其他种类的小型哺乳动物。小鼠腹腔注射0.025%秋水仙碱溶液0.5 ml， 1h后处死。股骨被迅速剥离，骨骺被移除，骨髓被从皮下注射器中取出的温的低渗柠檬酸钠溶液冲洗出来。轻轻地将骨髓吸进和吸出注射器，将其转化为精细的悬浮液。悬浮液在37℃的柠檬酸盐溶液中保存10分钟。结缔组织和骨针状体在细菌过滤管中通过尼龙螺栓布离心去除，然后在醋酸酒精（1:3）中固定并通过标准Feulgen程序染色。每次更换试剂后，细胞通过缓慢离心进行浓缩。强调了秋水仙碱预处理和细胞悬浮液处理的优点。
The method was developed for bone marrow of mice but is applicable to other tissues and other species of small mammals. Mice are injected intraperitoneally with 0.5 ml of 0.025% colchicine solution and killed 1 hr afterwards. The femurs are dissected out rapidly, the epiphyses are removed, and the marrow is washed out of the shafts by warm hypotonic sodium citrate solution from a hypodermic syringe. Gentle aspiration of the marrow into and out of the syringe converts it into a fine suspension. The suspension is kept in the citrate solution at 37[degree]C for 10 min. Connective tissue and bony spicules are removed by centrifuging through Nylon bolting cloth in a bacterial filtration tube, before fixing in acetic-alcohol (1:3) and staining by the standard Feulgen procedure. The cells are concentrated for each change of reagent by centrifuging slowly. The advantages of colchicine pretreatment and of working with cell suspensions are emphasized.