戊二醛置换法对大鼠冷冻保存肾脏超微结构的影响  

The Effect of Glutaraldehyde Replacement Method on Ultrastructure Changes of Cryo-preserved Kidney in Rats

作　　者：王红[1] 赵秀娟[1] 徐晨[1] 罗子国[1] 

机构地区：[1]重庆医科大学生命科学研究院电子显微镜室,重庆400016

出　　处：《实验室研究与探索》2011年第1期45-47,共3页Research and Exploration In Laboratory

摘　　要：探讨戊二醛置换法在不同复温方法下对液氮低温保存大鼠肾脏超微结构的影响。新鲜大鼠肾脏组织液氮保存3个月后,随机分为2组采用不同的复温方法。第1组标本直接放入4%的戊二醛溶液中在4℃冰箱内固定24 h复温;第2组标本先置入-80℃冰箱复温72 h,再放入4%的戊二醛溶液中在4℃内固定24 h复温。2组标本经常规透射电镜样品制备后,电镜下对超薄切片进行观察。结果表明,第1组样品中出现肾小球和肾小管细胞核异染色质边集、部分线粒体内室肿胀;第2组肾小球和肾小管细胞超微结构保存良好。提示低温保存的大鼠肾脏标本于-80℃冰箱复温72 h后,再置入4%的戊二醛溶液中在4℃内复温24 h,可以良好地保存细胞超微结构。结果表明,采用梯度复温戊二醛固定液置换法是液氮低温冷冻固定组织标本进行超微结构观察的一种简便易行的有效复温固定方法,对临床和研究室开展电镜研究具有实际的应用价值。The puopose of this paper is to investigate the effect of the glutaraldehyde replacement method on ultrastructure of cryo-preserved kidneys in rats with different rewarming methods.After cryo-preserved in liquid nitrogen for 3 months,the kidneys of rats were randomly divided into two groups with different rewarming methods.In the first group,samples were directly fixed in 4% glutaraldehyde at 4 ℃ for 24 hours for rewarming.In the second group,samples were put into-80 ℃ refrigerator for 72 hours for rewarming at first,and then fixed in 4% glutaraldehyde at 4 ℃ for rewarming for 24 hours.The ultrathin section was prepared and observed under the transmission electron microscope.Some mitochondria swelling and chromatin margination was found in glomerulus and renal tubular in the first group samples.However,in the second group,the ultrastructure of glomerulus and renal tubular are normal.Only very few mitochondria swelling can be observed.These results suggest that the preservation of the ultrastructure of rat kidneys with cryo-preserved in liquid nitrogen worked very well after rewarming at-80 ℃ for 72 hours and then fixed in 4% glutaraldehyde at 4 ℃ for 24 hours.This gradual rewarming method of glutaraldehyde replacement for the cryo-preserved samples is very simple and useful for the preservation of the ultrastructure of tissues after long storage in liquid nitrogen.It is also valuable for clinical electron microscopic studies.

关 键 词：肾脏 超微结构 冷冻保存 大鼠 

分 类 号：R446.61[医药卫生—诊断学] Q78[医药卫生—临床医学]