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目的寻找一种适合于遗传性视网膜色素变性的动物模型皇家外科学院大鼠(royal college of surgeons rat,RCS)眼球冰冻切片前的固定方法。方法 RCS大鼠5只,麻醉灌注后,5只眼行4%多聚甲醛固定1 h,30%蔗糖脱水过夜;5只眼用福尔马林、酒精、冰醋酸混合固定液(Davidson’s Fluid)固定12h后,以5%、10%、20%、30%蔗糖进行梯度脱水各4 h;再行眼杯OCT包埋后冰冻切片。结果 Davidson’s固定后的视网膜组织结构清晰,视网膜各层结构内的细胞排列紧密,无明显裂隙存在,且发生人为视网膜脱离概率低,无自发荧光干扰;相反,采用4%多聚甲醛固定的视网膜组织各层细胞排列较疏松,有一定的裂隙或空泡存在。结论 Davidson’s固定液适用于RCS大鼠视网膜组织的固定,效果优于单纯的4%多聚甲醛固定液。
Objective To find a suitable animal model of hereditary retinitis pigmentosa fixation before royal eyeball frozen section in royal college of surgeons rat (RCS). Methods Five rats of RCS were anesthetized and perfused. Four eyes were fixed with 4% paraformaldehyde for 1 hour and 30% sucrose was dehydrated overnight. Five eyes were infused with formalin, alcohol and Davidson’s Fluid, Fixed 12h, 5%, 10%, 20%, 30% sucrose gradient dehydration each 4 h; OCT OCT again after embedding frozen section. Results The structure of Davidson’s fixed retinal tissue was clear and the cells in retinal layers were closely arranged with no obvious fissure and the probability of artificial retinal detachment was low and there was no autofluorescence interference. On the contrary, 4% paraformaldehyde-fixed retinal tissue The layers of cells arranged more loose, there are some cracks or vacuoles exist. Conclusion Davidson’s fixative is suitable for immobilization of retinal tissue in RCS rats, and its effect is better than that of 4% paraformaldehyde fixative.