Evans Blue fluorescence permits the rapid visualization of non-intact cells in the perilesional rim of cold-injured rat brain.

A focal cold lesion-induced injury, i.e., a model of focal vasogenic brain edema, enhances the permeability of the blood-brain barrier and cell membrane in the perilesional rim. However, non-intact cells can be detected, e.g. by markers of apoptosis, only hours or even days after the injury. The early membrane dysfunction allows extravasated serum proteins to enter the injured cells, which can be readily visualized if the plasma albumin was previously bound to fluorescent tracers, such as Evans Blue (EB). The aim of this study was to demonstrate injured cells that take up the EB/albumin conjugate in the perilesional rim. This tracer was administered 3.5 h after the induction of the injury and the animals were sacrificed 30 min later. With an excitation wavelength of 530-550 nm, the EB-positive cells emitted bright-red fluorescence at > 590 nm and were easy to count. No positive cells were observed in the controls. This method provides more information than the classical 2,3,5-triphenyltetrazolium chloride reaction, because it permits an assessment of the density and distribution of cells with non-intact cell membranes in the perilesional area following cerebrocortical injury.