比较4种不同复合固定液对小鼠视网膜的固定效果  

Fixation effects of four different complex fixatives on mouse retina

作　　者：裴希 吴秋萍[2] 柯晓云[1] 彭艳阳 陈晓红[1] 

机构地区：[1]南方医科大学珠江医院眼科,广东省广州市510280 [2]南方医科大学珠江医院儿科,广东省广州市510280

出　　处：《中国组织工程研究》2014年第24期3791-3796,共6页Chinese Journal of Tissue Engineering Research

基　　金：广东省科技计划项目(2011B031800105)资助~~

摘　　要：背景:在各种实验与临床病理研究中,需要一种能够较好固定视网膜并且避免固定液性视网膜脱离的眼球固定液。目的:比较4种不同复合固定液对小鼠视网膜的固定效果。方法:40只昆明小鼠处死后,立即摘除眼球,并将其随机分为4组,每组20只眼球。分别浸泡在固定液1(多聚甲醛4 g,加入0.1 mol/L磷酸缓冲液80 mL,加热至60℃左右,持续搅拌,使粉末完全溶解,加少许1 mol/L NaOH澄清。待冷却后,加冰醋酸5 mL、丙酮10 mL,用0.1 mol/L磷酸缓冲液补足至100 mL),固定液2(5%重铬酸钾40 mL、体积分数为10%甲醛10 mL、冰醋酸10 mL、三氯醋酸10 mL、10%醋酸钠10 mL);固定液3(冰醋酸10 mL、体积分数为40%甲醛溶液20 mL、生理盐水70 mL、体积分数为75%乙醇10 mL混合);固定液4(无水乙醇60 mL、体积分数为40%甲醛溶液10 mL、冰醋酸10 mL、氯仿20 mL)。固定24 h进行常规脱水、石蜡包埋固定、苏木精-伊红染色。结果与结论:经过固定液4固定的视网膜组织可以清晰看到视网膜各层,而且罕见固定液性视网膜脱离。由于固定液4中加入能与乙醇、冰醋酸互溶的氯仿,有效地促进了固定液的眼球渗透,故降低了固定液性视网膜脱离,效果比其他几种固定液好。BACKGROUND： An optimal fixative, which can avoid retinal detachment, has been the subject of intensive experiments and clinical pathological studies. OBJECTIVE： To compare the fixation effects of four different complex fixatives on mouse retina. METHODS： Forty Kunming mice were sacrificed, and the eyeballs were enucleated and divided randomly into four groups, with 20 eyeballs in each group. The eyeballs in each group were respectively fixed in Fixative 1 （4 g paraformaldehyde were added into 0.1 mol/L phosphate buffer 80 mL, heated to 60 ℃, agitated-still the powder were dissolved, then clarified with 1 mol/L NaOH. When it was cooled down, 5 mL glacial acetic acid and 10 mL acetone were added, and 0.1 mol/L phosphate buffer was added to make up the volume to 100 mL）, Fixative 2 （5% potassium dichromate solution 40 mL, 4% formaldehyde 10 mL, glacial acetic acid 10 mL, trichloroacetic acid solution 10 mL, 10% sodium acetate trihydrate solution 10 mL）, Fixative 3 （glacial acetic acid 10 mL, 40% formaldehyde 20 mL, physiological saline 70 mL, 75% alcohol 10 mL）, and Fixative 4 （alcohol 60 mL, 40% formaldehyde 10 mL, glacial acetic acid 10 mL, chloroform 20 mL）. After 24 hours of fixation, paraffin section and hematoxylin-eosin staining were performed.RESULTS AND CONCLUSION： The retina fixed by Fixative 4 （alcohol 60 mL, 40% formaldehyde 10 mL, glacial acetic acid 10 mL, chloroform 20 mL） gave us a clear view of every layer of the retina, and retinal detachment was rarely seen. Fixative 4 promotes the permeation into the eyeballs, reduces the incidence of retinal detachment, and is better than the other fixatives.

关 键 词：组织构建 组织工程 复合固定液 小鼠 视网膜 组织固定 

分 类 号：R318[医药卫生—生物医学工程]