Citation:Liang XL,Li J,Chen F,Ding XY,Yang XX,Long LX.A comparing study of quantitative staining techniques for retinal neovascularization in a mouse model of oxygen-induced retinopathy.Int J Ophthalmol 2012;5(1):1-6,doi:10.3980/j.issn.2222-3959.2012.01.01
A comparing study of quantitative staining techniques for retinal neovascularization in a mouse model of oxygen-induced retinopathy
  
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DOI:10.3980/j.issn.2222-3959.2012.01.01
Key Words:neovascularization  endothelial cell  CD31, hematoxylin-eosin staining  immunohistochemistry  immuno- fluorescence  retinopathy of prematurity
Fund Project:Supported by National Natural Science Foundation of China (No. 30973899)
                 
AuthorInstitution
Xiao-Ling Liang Key Lab of Ophthal Ministry of Edu, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou , Guangdong Province, China
Jie Li Key Lab of Ophthal Ministry of Edu, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou , Guangdong Province, China
Fang Chen Key Lab of Ophthal Ministry of Edu, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou , Guangdong Province, China
Xiao-Yan Ding Key Lab of Ophthal Ministry of Edu, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou , Guangdong Province, China
Xiu-Xia Yang Key Lab of Ophthal Ministry of Edu, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou , Guangdong Province, China
Liao-Xu Long Guangzhou Medical University, 195#, Dongfeng Xi Road, Guangzhou, Guangdong Province ,China
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Abstract:
      AIM: To explore an efficient, practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR).

    

    METHODS: Thirty C57BL/6J mice were explored in OIR model procedure. Eyes were removed for different staining methods including: (1) HE staining; (2) immunohistochemistry with Griffonia Simplicifolia Lectin(GSL);(3) Immunofluore- scence with FITC labeled CD31 antibody; (4) Two-step immunofluorescence with purified-CD31 antibody; (5) FITC-Dextran perfusion combined with two-step purified- CD31immunofluorescence. Images of the retinal vasculature were analyzed by imaging software.

    

    RESULTS: GSL immunohistochemistry could clearly demonstrate the deep and superficial capillary beds. FITC labeled CD31 Immunofluorescence was blurring with high fluorescence background which was hard to distinguish retinal neovascularization in some area. Excellent detail of neovas- cularization and preexistent retinal vessels was provided in two-step Purified-CD31 immunofluorescence group.

    

    CONCLUSION: GSL immunohistochemistry can clearly demonstrate neovascularization tufts in deep and superficial capillary beds. Immunofluorescence of specific antigen CD31 on vascular endothelium can selectively label the neovascularization of mouse retina. When combined with computer analysis software, it is an effective and objective quantitative method to evaluate the retinal neovascularization in OIR mouse model.

    

PMC FullText Html:http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3340848/
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