小鼠整体胚胎和新生鼠石蜡切片技术的改进

目的探讨小鼠整体胚胎和新生鼠石蜡切片技术的改进。方法收集不同胚龄和日龄的小鼠整体胚胎及新生鼠14.5、15.5及16.5 d的胚胎,清洗后直接置于4%中性甲醛中固定24 h以上;胚龄17.5、18.5 d及日龄0、1、2 d的小鼠,先用注射器往胸腹部及脑部缓慢注入固定液,再整体固定于标本瓶中24 h以上。经全自动密闭式组织脱水机编程脱水,石蜡包埋技术制备小鼠整体胚胎和新生鼠切片。应用HE染色方法对切片进行染色检验,光学显微镜下观察小鼠整体胚胎和新生鼠的不同组织结构。结果使用全自动密闭式组织脱水机处理小鼠整体胚胎和新生鼠,经过特定的脱水程序处理,镜下观察不同胚龄及日龄的不同组织器官,结构完整,层次清晰,一致性好。结论本实验室利用全自动密闭式组织脱水机对小鼠胚胎和新生鼠进行处理,通过设置合理程序,能够同时处理不同胎龄和日龄的小鼠整体胚胎及新生鼠;脱水后制备的石蜡切片结果稳定,一致性好,为小鼠遗传学和发育学研究提供了良好的技术支持。

Technical improvement of the paraffin sections in mouse embryos and newborn

Objective To improve the method of paraffin section preparation of whole mouse embryos and newborn mice. Method Specimens of mouse embryos and newborn mice were collected. Embryos aged 14.5, 15.5 and 16.5 days were directly placed in 4% neutral formaldehyde and fixed for more than 24 hours. For embryos aged 17.5 and 18.5 days, and 0-, 1- and 2-day old newborn mice, a small amount of formaldehyde was slowly injected into the chest, abdomen and brain at first, respectively, and then the whole specimens were fixed for more than 24 hours. Tissues were processed together using a fully-enclosed tissue processor. Paraffin-embedded slices were observed under microscope after HE staining. Results The quality of specimens following the improved method was better than before. The structure of different tissues from both mouse embryos and newborn mice can be easily determined. No fragmentation was found in any organ tissue under the light microscope. Conclusions We can handle whole body specimens of both mouse embryos and newborn mice simultaneously by adjusting the program of the fully-enclosed tissue processor. The staining results of the sections which were dehydrated using this improved method are clear and stable. This improved method may provide a useful approach in research of genetics and developmental biology.