Evans blue-albumin tracing for pulmonary transvascular permeability was conducted as previously described [3 (link)]. The right jugular vein of ketamine-xylazine anesthetized mice was injected with 8 µl/g body weight Evans Blue–albumin (40 mg/ml BSA containing 1% Evans blue dye, Sigma) and left to circulate for 45 min. A 2-min PBS perfusion into the right ventricle was used to wash out the intravascular Evans Blue. Lungs were excised, homogenized in PBS (1 ml), and extracted using formamide (2 ml) at 60°C overnight. A Shimadzu 1650 PC UV-vis spectrophotometer (Kyoto, Japan) was used to measure Evans Blue content at OD620 and normalized against body weight.
Protocol detail
Pulmonary Transvascular Permeability Measurement
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Albumin
Body weight
Evans blue
Formamide
Jugular vein
Ketamine
Lungs
Mice
Perfusion
Permeability
Right ventricle
Xylazine
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Variable analysis
independent variables
- Injection of Evans Blue–albumin (8 µl/g body weight)
- Evans Blue content in lung tissue (measured by UV-vis spectrophotometry at OD620 and normalized against body weight)
- Ketamine-xylazine anesthesia
- 2-min PBS perfusion into the right ventricle to wash out the intravascular Evans Blue
- Lung tissue homogenization and extraction using formamide
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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