Abstract: Using the technology of immunohistochemical staining and color image analysis to investigate the spatiotemporal pattern and age_related changes of type Ⅰ collagen expression in the corneal development of Bufo raddei larvae during metamorphosis.Results showed that the inner and outer cornea combined together into mature cornea during metamorphosis.Both corneal epithelium and corneal stroma become thickened with progressively increase of the age.The analysis of immunohistochemical staining by color image analysis found that type Ⅰcollagen expressed widely during the period,existing in Bowman's membrane,Descemet's membrane,corneal stroma and endothelium.Espeacially,a progressively change in corneal stroma was observed.ConclusionThe defect of qualitative index may be remedied by color index in the quantitative immunohistochemical image analysis of type Ⅰ collagen.The inner and outer cornea combine together gradually with type Ⅰ collagen increasing in the corneal development of Bufo raddei larvae during metamorphosis.Type Ⅰcollagen,which is the main component in the increased collagens,expresses progressively with age.Type Ⅰcollagen may be the important component in corneal stroma,and it plays an important role in the construction and maintaining of the corneal stroma.

Key words: Bufo raddei larvae, Cornea development, Type Ⅰ collagen, Immunohistochemistry, Image analysis

摘要： 应用彩色图像分析技术对Ⅰ型胶原的免疫组化彩色图像分析可以弥补单纯应用定性指标或形态学指标的不足。花背蟾蜍蝌蚪在变态期角膜发育过程中随着Ⅰ型胶原的增多,内、外角膜逐渐愈合。Ⅰ型胶原的表达存在增龄变化,可以推测Ⅰ型胶原是构成角膜基质的主要成分,对建立和维持角膜支架有重要作用。

关键词: 花背蟾蜍蝌蚪, 角膜发育, Ⅰ型胶原, 免疫组化, 图像分析