关键词: 异硫氰酸荧光素, 透明, 整体标记, 冷冻切片, 小鼠

Abstract: Objective To develop whole-mount tissue clearing methods for the cutaneous microvessel in situ visualization. Methods Twenty ICR mice were randomly divided into 2 groups: Ⅰ and Ⅱ. Our approach included labeling luminal surfaces of endothelial cells by i.v injection of fluorescein isothiocyanate(FITC) conjugated Anti-CD31, ulex europaeus agglutinin-I(UEA-I) respectively, and then subdivided them into unclearing group (a) and clearing group (b). The former was observed using a fluorescence microscope after frozen section, while the latter was observed followed by optical clearing of whole-mount tissue to reduce light scatter and permit visualization of microvessel morphology. Results GroupⅠa andⅡa had lower vessel density, the angiosomes were damaged or destroyed obviously；GroupⅠb andⅡb presented with limpid and integrity angiosomes. This optical clearing technique allows imaging of the microvessel by microscopy, to depths of up to 1 000μm below the specimen surface. Conclusion The application of intravital fluorescent staining wit

Key words: Fluorescein isothiocyanate, Clearing, Whole-mount labeling, Frozen section, Mouse