关键词: 关键词 , 冷冻超薄切片 免疫电镜 , 肾活检 , Tokuyasu技术 胶体金 , LR White树脂

Abstract:

ABSTRACT Objective: To investigate the application of ultrathin cryo-immunogold electron microscopy methods in renal biopsy. Methodology: Tissue blocks of renal biopsy were fixed in fixation solution, and further sucrose cryoprotected. Ultrathin cryosections were cut with the application of improved technology of Tokuyasu. The grids were immunolabelled using IgA, ?, ?, C3aR, podocine, and nephrin. Combined with immunofluorescence markers, immunoelectron microscopy for routine Epon812 and LR White embedding immunoelectron microscopy at low temperature as a reference, ultrathin cryosections were observed and the affects of those methods on the ultrastructure of renal biopsy, antigen preservation and immunolabelling were compared. Results: The ultrathin cryosections of renal tissue well preserved, fine structure. The intracellular structure was also clear while intracellular antigens preservation was good. However, the intracellular matrix, stored glycogen was not ideal. IgA 、?、? immune markers of gold particles density was high, distribution specificity and high sensitivity. The antigens of C3aR, podocine and nephrin , which might be difficult to detected by Epon812 routine immune electron microscopy and LR White embedding immunoelectron microscopy, were well marked, and the immune markers of gold particle showed high density and distribution of specific and sensitive. Conclusion: The immunogold labeling of renal biopsy ultrathin cryosections was specific and high sensitive. This technique was a more satisfactory method at the ultrastructure level of renal morphological changes.

Key words: Key words: ultrathin cryosections , Immunoelectron microscopy  , renal biopsy , Tokuyasu technique Immunogold  , LR White resin