实验用斑马鱼成鱼内脏组织病理学检查方法的优化

摘要

Objective To explore the optimized method ofparatfin serial section and hematoxylineosin (HE) staining of the adult zebrafish visceral tissues.Methods After the zebrafish was fixed,the tissues were segmented,and the thoracic/abdominal segment which contains internal organs was stripped of skin and dehydrated by gradient ethanol (dehydration time was optimized to 30 min),transparented by xylene and embedded by paraffin.The tissue was sectioned serially along the body axis,rinsed with warm water after HE staining,and observed under microscope.Results The microscopic observation of heart,liver,spleen,gallbladder,kidney,pancreas,gastrointestinal tract,testis and ovary showed that effects of tissue sectioning and staining were favorable.The sections were intact without collapse or fracture,the anatomical and cytological structure of all organs were clear and complete,the nucleus were neatly blue-stained and the cytoplasm were neatly red-stained.Conclusion With the optimized histopathological method,we were able to obtain high quality histological specimens of the internal organs of adult zebrafish.%目的 探索并优化适合于斑马鱼咸鱼内脏组织的病理学检查技术.方法 将斑马鱼整条固定于Dietrich's固定液后,分段切开,取包含内脏组织的胸腹段,剥除斑马鱼鱼皮,梯度乙醇脱水时间优选为30 min、二甲苯透明、石蜡包埋、沿斑马鱼体轴纵向连续切片、HE染色后用温水冲洗切片进行颜色反蓝,正置显微镜下观察效果.结果 经过反复的条件摸索和优化,光学显微镜下观察心脏、肝脏、脾脏、胆囊、肾脏、胰腺、消化道、精巢和卵巢等组织切片和染色的效果,可见斑马鱼组织均切片完整,无折叠或断裂,各脏器的解剖学与组织细胞学结构清晰,易于辨认,细胞核与细胞质染色分明.结论 优化后的斑马鱼咸鱼内脏组织病理学方法效果较为理想,可获得高质量的内脏组织学标本.