Original articleRe-evaluation of Evans Blue dye as a marker of albumin clearance in murine models of acute lung injury
Section snippets
Chemicals
Tetrasodium salt of EB (tetrasodium 4-amino-6-[4-[4-(8-amino-1-hydroxy-5,7-disulfonato-naphthalen-2-yl)diazenyl-3-methyl-phenyl]-2-methyl-phenyl]diazenyl-5-hydroxy-naphthalene-1,3-disulfonate; MW 960.8), bacterial LPS (serotype O127:B8), and bovine serum albumin (BSA) (fraction V, low-endotoxin; MW 66,000) were from Sigma Chemical Co. (St. Louis, Mo). All other chemicals were analytical or cell-culture grade and were obtained from various commercial sources. Evans Blue dye conjugated to albumin
A lung-specific tissue-specific correction factor
To initially test whether a correction factor (as in Linderkamp et al18) can at all be obtained from mouse lung homogenates, we recorded absorbances at both 620 nm and 740 nm of such homogenates prepared as described in the Materials and Methods section, and we calculated the least-squares fit between the paired absorbance recordings for each sample (Fig 1). We found that a significant fit (P < 0.01) could be obtained even with a small sample size (N = 18), in contrast to the large N (147)
Discussion
These data clearly demonstrate that the widely used EB dye extravasation method to quantify barrier dysfunction can be improved in terms of repeatability and sensitivity by using tissue-specific turbidity correction factors. One clue as to why that is so may be found by comparing the commonly used serum correction factor18 with the lung-specific ones experimentally determined in this manuscript. Such a comparison shows that, although the slopes are not significantly different, the intercepts of
References (27)
- et al.
Bestimmung der bindungskapazität des albumins für Evans-Blue (T 1824) mit der freien präparativen elektrophorese nach barrollier
Clin Chim Acta
(1960) - et al.
Evans blue fluorescence: quantitative and morphological evaluation of vascular permeability in animal tissues
J Neurosci Meth
(1983) - et al.
Evaluation of arterialized vein graft permeability with Evans Blue dye and iodine 125-labeled albumin
Ann Thorac Surg
(1989) - et al.
Aquaporin channels may modulate ventilator-induced lung injury
Resp Physiol
(2001) The relationship of extinction to wave-length in turbid sera and other suspensions
J Biol Chem
(1943)- et al.
Attenuation by intravenous 2-chloroadenosine of acute lung injury induced by live escherichia coli or latex particles added to endotoxin in the neutropenic state
J Lab Clin Med
(2003) - et al.
Polynitroxyl albumin inhibits inflammation and vasoocclusion in transgenic sickle mice
J Lab Clin Med
(2005) - et al.
Evans blue dye as a marker of albumin clearance in cultured endothelial monolayer and isolated lung
J Appl Physiol
(1992) In vivo vascular leakage assay
- et al.
Clinical studies of the blood volumeI. Clinical application of a method employing the azo dye “Evans Blue” and the spectrophotometer
Clin Invest
(1937)
The binding of T-1824 and structurally related diazo dyes by the plasma proteins
Am J Physiol
Equilibrium and kinetic properties of the Evans Blue-albumin system
Am J Physiol
Combination of Evans Blue with plasma protein: its significance in capillary permeability studies, blood dye disappearance curves, and its use as a protein tag
Am J Physiol
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Supported by the NIH/NHLBI Grant P50 HL 73994 (to J.G.N.G.).