To quantify vascular leakage, Evans blue (Sigma-Aldrich) was injected i.v. at a concentration of 20 mg/kg body weight diluted in PBS. After 4 h, brain and liver tissue were removed after perfusing mice with saline. Evans blue was extracted from tissues using N,N-dimethyl formamide (Sigma-Aldrich) as described previously (Kim et al., 2009 (link)). All samples were plated in triplicate on black 96-well flat-bottom plates (Corning), and fluorescence emission was quantified using a Varioskan Flash fluorometer (620-nm excitation; 695-nm emission; Thermo Fisher Scientific).
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Quantifying Vascular Leakage via Evans Blue
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Other organizations : National Institutes of Health
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Variable analysis
independent variables
- Injection of Evans blue at a concentration of 20 mg/kg body weight
- Fluorescence emission of Evans blue extracted from brain and liver tissue
- Dilution of Evans blue in PBS
- Perfusion of mice with saline
- Use of black 96-well flat-bottom plates
- Quantification of fluorescence emission using a Varioskan Flash fluorometer (620-nm excitation; 695-nm emission)
- No positive or negative controls were explicitly mentioned.
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