Assessment Of Leukocyte And Platelet Adhesion

Adhesion of leukocytes and platelets following ischemia and reperfusion can be monitored in vivo using intravital video microscopy. The head of the animal is immobilized and a hole drilled through the skull using a high-speed micro drill (1 mm posterior from bregma and 4 mm lateral from the midline). The dura matter is not cut because the fluorescently labeled blood cells are easily observed and intracranial pressure is well maintained with the dura matter intact. Artificial cerebrospinal is placed on the exposed brain tissue. For focal ischemia models, the observation area includes the infarcted region following MCAO. For global ischemia models the observation area includes the entire cerebral cortex. Platelets are isolated from a donor animal using a series of centrifugation steps and labeled ex vivo with carboxyfluorescein diacetate succinimidyl ester. Once the platelet data is collected, endogenous leukocytes are labeled in vivo by infusing rhodamine-6G (100 ml; 0.02%) over five minutes and allowed to circulate an additional five minutes before observation. An upright Nikon microscope equipped with a SIT camera (C2400-08; Hamamatsu Photonics) and a mercury lamp is used to observe the cerebral microcirculation. The images are received by a CCD video camera and recorded on a video recorder equipped with a time-date generator (WJ-810; Panasonic).

Blood Pressure Health

Blood Pressure Health

Your heart pumps blood throughout your body using a network of tubing called arteries and capillaries which return the blood back to your heart via your veins. Blood pressure is the force of the blood pushing against the walls of your arteries as your heart beats.Learn more...

Get My Free Ebook

Post a comment