The mice were anesthetized, and the femoral straight muscle venules were exposed by a resection in the anterior part of the thigh for exhibition of the femoral straight muscle. The animals received an i.v. injection from Rodamin 6G (0.3 mg kg–1, Sigma-Aldrich, Germany) to fluorescent labeling of leukocytes. An intravital microscope (ECLIPSE 50i; Nikon) with a 20 objective lens was used to examine the muscle microcirculation. A digital camera (DS-Qi1MC; Nikon) was used to acquire the images that were recorded for playback analysis with Nikon imaging software. The counting of rolling and adherent leukocytes was realized according to our previously published method (Rezende et al., 2017). Rolling leukocytes was defined as those cells moving thought the observed field at a velocity less than that of erythrocytes within a given vessel during 1 min. Leukocyte was considered to be adherent if it remained stationary for at least 30 s, and total leukocyte adhesion is quantified as the number of adherent cells within a 100 μm length of venule in 1 min.

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