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本文介绍了用鱼类(鲑鱼)胚胎制作永久性染色体标本的方法。其程序如下: 1.在解剖镜下,在林格(Ringer)氏液里用解剖刀划破卵膜,再用弯头摄子从正在发育的卵中取出胚胎。2.用林格氏液清洗样本,除去粘性卵黄物质和小油滴。3.在室温条件下用0.1%胰蛋白酶(克/单位体积林格氏液)处理10分钟。4.在0.075克分子氯化钾溶液中将样本透析20～30分钟。5.用卡诺(Carnoy)氏液(甲醇3份,醋酸1份)至少固定20分钟。6.将样本置于清洁的载玻片上,再加一滴(2～4微升)蒸馏水以防干燥。7.用锋利的解剖刀对胚胎细胞进行切片,制成乳状细胞悬液。8.在载玻片上滴约40微升卡诺氏液,使细胞悬液扩展到整个片子上。
This article describes a method of making permanent chromosome specimens from fish (salmon) embryos. The procedure is as follows: 1. Under a dissecting microscope, scratch the egg membrane with a scalpel in a Ringer’s solution and remove the embryo from the developing egg using elbow picks. 2. Rinsate the sample with Ringer’s solution to remove sticky yolk material and small oil droplets. 3. Treat with 0.1% trypsin (g / unit Ringer’s solution) for 10 minutes at room temperature. 4. Dialyze the sample in 0.075 mM potassium chloride solution for 20-30 minutes. 5. Use Carnoy’s solution (3 parts of methanol and 1 part of acetic acid) for at least 20 minutes. 6. Place the sample on a clean glass slide and add a drop of (2 to 4 μl) distilled water to prevent drying. 7. Use a sharp scalpel to slice the embryonic cells and make a suspension of the milky cells. 8. Dilute approximately 40 μl of Cannott’s solution to the slide to allow the cell suspension to spread over the entire slide.