[关键词]
[摘要]
目的:为解决豚鼠眼球组织切片制备过程中所存在的视网膜脱片问题,采用不同固定方法,优化固定效果。
方法:2周龄正常豚鼠(75只)随机分为5个大组(A-E组),15个小组,每小组5只。A组(1-3小组)眼球分别置于FAS、Davidson固定液1(D1)和Davidson固定液2(D2)中固定24 h; B组(4-6小组)眼球在固定液中固定1 h后剪切角膜,再固定2 h; C组(7-9小组)眼球在固定液中固定1 h后沿视神经方向将眼球分为左右两半,再固定2 h; D组(10-12小组)眼球在固定液中固定3 h后将眼球分为左右两半; E组(13-15小组)眼球在固定液中固定3 h后剪切角膜。经苏木精-伊红(HE)染色比较各个小组眼球后极部固定效果。
结果:形态观察表明1-6小组、11-15小组眼球表面光滑圆润,色泽透明,7-10小组眼球凹陷皱缩变形。HE染色表明大部分组别眼球后极部组织切片卷曲缠绕,视网膜脱离; 1、5、6、14、15小组切片结构规整,其中14小组形态最佳,视网膜、脉络膜、巩膜连接紧密,组织结构清晰,细胞排列规整。
结论:采用D1固定液固定3 h后剪切角膜的固定效果最为理想,适用于豚鼠眼球后极部相关组织研究。
[Key word]
[Abstract]
AIM: To avoid the problem of retinal dissection in guinea pig large eyeball tissue sections, different methods were used to optimize the fixation effect of the posterior pole of the eyeball.
METHODS: A total of 75 normal guinea pigs(2 weeks old)were randomly divided into 5 large groups. Group A(1-3 small groups), the entire eyeball was fixed with FAS, Davidson fixative 1(D1), and Davidson fixative 2(D2)for 24 h; group B(4-6 small groups), the entire eyeball was fixed with FAS, D1, and D2 for 1 h, then cut the cornea and fix it in their respective fixatives for 2 h; group C(7-9 small groups), the eyeball was fixed in FAS, D1, and D2 for 1 h, divided into left and right halves along the direction of the optic nerve, and then placed them in their respective fixation solutions for 2 h; group D(10-12 small groups), after fixation for 3 h in FAS, D1, and D2, the eyeball was divided into left and right halves along the optic nerve direction; group E(13-15 small groups), the cornea was cut after fixation for 3 h in FAS, D1, and D2. Hematoxylin-eosin(HE)staining was used to compare the fixation effect on posterior eyeball in each group.
RESULTS: After fixation, the surface of the eyeballs in groups, 1-6 and 11-15 was smooth and round, with a transparent and bright color. In groups 7-10, the eyeballs were sunken, wrinkled, and deformed. The HE staining showed that the eyeball morphology of groups 1, 5, 6, 14, and 15 was significantly better than the other groups, with a regular internal tissue structure. The eyeballs of the other groups were sunken and wrinkled, and the internal tissue was curled and tangled, with severe retinal detachment. In groups 1, 5, 6, 14, and 15, the retina, choroid, and sclera tissues of group 14 were closely connected, without obvious retinal detachment, rupture, or curling. The tissue structure was clear and visible, and the cells were arranged neatly.
CONCLUSION: The fixation effect of cutting the cornea after fixing guinea pig eyeball with D1 fixative for 3 h is the most ideal, and this operation method is simple and suitable for studying the related structures of the posterior pole of the eye.
[中图分类号]
[基金项目]
山东省重点研发计划项目(No. 2021LCZX09); 山东省自然科学基金项目(No.ZR2021LZY045); 山东省中医药科技项目(No.2021Q096,Q-2023015); 山东省医药卫生科技发展计划(No.202207020876)