›› 2010, Vol. 41 ›› Issue (5): 774-776.doi: 10.3969/j.issn.0529-1356.2010.05.027

• 技术方法 • Previous Articles    

Application of intravital fluorescent staining with tissue clearing in displaying microvascular architecture of mouse skin

  

  1. Department of Anatomy, Wenzhou Medical College, Zhejiang Wenzhou325035, China
  • Received:2009-09-17 Revised:2009-11-02 Online:2010-10-06
  • Contact: TANG Mao-lin

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

CLC Number: