耳蜗组织低温包埋切片技术

Cryoembedding and Sectioning of Cochleas

目的探索一种新的耳蜗组织切片方法,以获得高质量的切片,满足其免疫组织化学染色的需要。方法取1天龄大鼠、成年豚鼠、成年小鼠耳蜗标本经蔗糖快速浸透,用离心管定向并用新型组织包埋剂Tissue-Tek(OCT)包埋,液氮中快速冷冻后在恒冷箱切片机中切片,然后分别做苏木素、伊红染色和免疫组化染色,观察组织切片的形态结构。结果经免疫荧光和组织化学染色后组织切片形态结构完整,耳蜗基底膜平直,盖膜及前庭膜完整,毛细胞与支持细胞间结构完整,细胞核清晰。结论耳蜗组织免疫组化染色应选择低温包埋切片技术制作的切片。

Objective To explore a new method of cochlear sectioning to get high qualitative cochlear section catering for immunohistochemistry of cochlea. Methods The cochlear samples cochlea of guinea pig and mouse were infiltrated with increasing concentrations of sucrose, followed by degassed embedding medium before final orientation and freezing. The sections were stained by hematein, eosin and immunohis tochemistry. The morphology of the tissue sections was observed. Results After immunofluorescence and histochemistry staining, the overall intergrity of the tissue section could be preserved, including basilar membrane, tectorial membrane and Reissner＇ s membrane, hair cell and supporting cells. The nuclei were very distinct. Conclusion Sectioning of cochlear inununohitochemistry should utillize cryoembedding and sectioning technique to preserve overall structure and cellular resolution.