An improved method for preparation of mouse retinal cryosections
An improved method for preparation of mouse retinal cryosections
复制标题
小鼠视网膜冷冻切片制备方法的改进
DOI：
10.4081/ejh.2020.3154
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发表时间：
2020-01-01
影响因子：
2
通讯作者：
Zhang, Houbin
中科院分区：
文献类型：
--
作者：
Li, Ling;Lin, Yongqiong;Zhang, Houbin
使用小鼠视网膜冷冻切片的免疫组织化学是视觉研究中使用的常规测定。然而，视网膜组织是脆弱的，并且难以获得理想的视网膜冷冻切片。在这里，我们开发了一种改进的方法制备视网膜冷冻切片。在从未固定或PFA固定的小鼠眼球中取出角膜和透镜之前，将Super Glue应用于巩膜表面。新方法在很大程度上防止了小鼠视网膜冷冻切片中的视网膜脱离。免疫染色视网膜冷冻切片来自PFA固定的小鼠眼睛使用杆和锥标记产生高质量的免疫荧光图像。使用纤毛特异性标记从未固定的小鼠眼睛获得的视网膜冷冻切片的免疫标记具有改善的感光体连接纤毛的方向。这种新方法大大改善了固定和未固定的小鼠眼睛的形态和免疫染色结果。
Immunohistochemistry using mouse retinal cryosections is a routine assay used in vision research. However, retinal tissues are fragile, and it is difficult to obtain an ideal retinal cryosection. Here, we developed a modified method for preparing retinal cryosection. Super Glue was applied on the surface of the sclera before the cornea and the lens are removed from either the unfixed or PFA-fixed mouse eyeballs. The new methods largely prevented retinal detachment in mouse retinal cryosections. Immunostaining of retinal cryosections derived from PFA-fixed mouse eyes using rod and cone markers yielded high-quality immunofluorescent images. Immunolabeling of retinal cryosections obtained from unfixed mouse eyes using a cilium-specific marker had improved orientations of photoreceptor connecting cilia. This new method substantially improves the morphology and immunostaining results of fixed and unfixed mouse eyes.