Evans blue

To make the indicator dye solution, Evans Blue (Sigma, MO) was dissolved in 0.9% sterile saline (Hospira, IL) to create a 1% solution. For i.t. experiments, the Evans Blue solution was administered at a rate of 0.5uL/hr for a period of 7 days via 1007D Alzet osmotic mini-pumps (Durect, CA). For s.c. experiments, the Evans Blue solution was delivered at a rate of 0.25uL/hr for a period of 7 days via 1002 Alzet osmotic mini-pumps (Durect, CA). All pumps were placed in 15mL conical tubes containing sterile 0.9% saline and primed overnight in a dry heat bath (Lab Armor, OR) at 37°C. Pumps for s.c., but not i.t., experiments were attached to bifurcated (bif.) catheters prior to priming.