Bx150wi

For intravital microscopy of the liver, overnight fasted animals were anesthetized via intraperitoneal (i.p.) injection with ketamine (100 mg/kg) and xylazine (10 mg/kg). A midline and left subcostal incision was made to exteriorize the liver. The hepatic ligaments were dissected and the intestine was covered with a saline-soaked gauze to minimize the tissue dehydration. The left liver lobe was then exteriorized and placed on a glass disc, and then the glass disc was covered with a glass slide for microcirculation analyses. With the use of a 10× ocular and 10× objective (Olympus BX150WI; Center Valley, PA, USA), images were displayed on a television monitor and recorded by a digital video recorder (DP73; Olympus, MA, USA) for offline analysis with the CellSens standard 1.9 software program (Olympus, Waltham, MA, USA). The leucocyte-endothelial interaction was evaluated by counting the number of labelled leucocytes (0.3 mg/kg rhodamine 6G, i.v.) rolling or adhering to sinusoids and postsinusoidal venules. Rolling leucocytes were defined as white blood cells with a slower velocity than the erythrocytes and with a detectable rolling motion. Leucocytes that remained stationary on the sinusoidal or venular endothelium for 30 s or longer were considered adherent cells. Rolling and adherent cells were counted in a 170-μm 2 area comprising sinusoids and postsinusoidal venules.