Measurement of vascular permeability in spinal cord using Evans Blue spectrophotometry and correction for turbidity

RE Warnick, JR Fike, PH Chan, DK Anderson… - Journal of neuroscience …, 1995 - Elsevier
RE Warnick, JR Fike, PH Chan, DK Anderson, GY Ross, PH Gutin
Journal of neuroscience methods, 1995Elsevier
Vascular permeability can be visualized by Evans Blue (EB) extravasation and quantified by
spectrophotometry after formamide extraction of the tissue. However, formamide extracts
show significant turbidity, which may contribute to the total optical density at the wavelength
of measurement (eg, 620 λ). We developed a simple method for estimating the component of
the total optical density of a dyed specimen contributed by turbidity. Our method, which uses
a determination of turbidity made at another point of the light spectrum (740 λ), was more …
Vascular permeability can be visualized by Evans Blue (EB) extravasation and quantified by spectrophotometry after formamide extraction of the tissue. However, formamide extracts show significant turbidity, which may contribute to the total optical density at the wavelength of measurement (e.g., 620 λ). We developed a simple method for estimating the component of the total optical density of a dyed specimen contributed by turbidity. Our method, which uses a determination of turbidity made at another point of the light spectrum (740 λ), was more precise than two other EB quantification techniques. We therefore recommend it for individual correction of formamide extracts of spinal cord specimens. The application of this technique to the brain remains to be determined.
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