Effect of acute coronary occlusion upon the movement of Evans blue dye into different areas of pig heart muscle

Caster, W. O.; Garamella, Joseph J.; Veloso, A.C. López; Naidu, R Purushotham

doi:10.1016/0002-8703(58)90208-4

Cited by 2 publications

(2 citation statements)

References 3 publications

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“…EB methodology, initially developed in the 1950’s for assessment of physiological albumin tissue distribution in mammals (4-6) became attractive for lung injury models in the 1980’s-1990’s, where EB could be quantified directly in lavage fluids and tissue extracts of larger organs (18-21). In addition, albumin-bound EB uptake into injured cardiomyocytes or neurons and subsequent quantitation by fluorescence microscopy has been exploited for cardiac and brain reperfusion after injury (22;23).…”

Section: Discussionmentioning

confidence: 99%

“…Vascular permeability in animal models of experimental inflammation is often based on measurement of albumin-bound Evans Blue (EB) extravasation into tissues and constitutes a highly relevant, physiological endpoint. However, measurement of EB in organs typically rely on a method that was developed in the 1950’s for the quantitation of albumin distribution in mammalian tissues (4-6). It requires drying of tissues and organic solvent extraction for days followed by absorbance (620-659 nm) measurements of EB in extracts.…”

Section: Introductionmentioning

confidence: 99%

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Infrared fluorescence for vascular barrier breach in vivo – A novel method for quantitation of albumin efflux

Drygalski¹,

Furlan-Freguia²,

Mosnier³

et al. 2012

Thromb Haemost

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Summary Vascular hyperpermeability contributes to morbidity in inflammation. Current methodologies for in vivo assessment of permeability based on extravasation of Evans Blue (EB)-bound albumin are cumbersome and often lack sensitivity. We developed a novel infrared fluorescence (IRF) methodology for measurement of EB-albumin extravasation to quantify vascular permeability in murine models. Vascular permeability induced by endotoxemia was examined for all solid organs, brain, skin and peritoneum by IRF and the traditional absorbance-based measurement of EB in tissue extracts. Organ IRF increased linearly with increasing concentrations of i.v. EB (2.5-25 mg/kg). Tissue IRF was more sensitive for EB accumulation compared to the absorbance-based method. Accordingly, differences in vascular permeability and organ EB accumulation between lipopolysaccharide-treated and saline-treated mice were often significant when analyzed by IRF-based detection but not by absorbance-based detection. EB was detected in all 353 organs analyzed with IRF but only in 67% (239/353) of organs analyzed by absorbance-based methodology, demonstrating improved sensitivity of EB detection in organs with IRF. In contrast, EB in plasma after EB administration was readily measured by both methods with high correlation between the two methods (n=116, r2=0.86). Quantitation of organ-specific EB-IRF differences due to endotoxin was optimal when IRF was compared between mice matched for weight, gender, and age, and with appropriate corrections for organ weight and EB plasma concentrations. Notably, EB-IRF methodology leaves organs intact for subsequent histopathology. In summary, EB-IRF is a novel, highly sensitive, rapid, and convenient method for the relative quantification of EB in intact organs of treatment versus control mice.

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