Animal models, particularly murine protocols on physical exercise, are widely used to investigate physiological adaptations and to study, prevent, and treat chronic non-communicable diseases. Controlled treadmill running enables precise manipulation of intensity and duration, overcoming limitations of voluntary wheel running for modeling inflammation. An incremental-speed treadmill test to fatigue disrupts homeostasis and acts as an acute inflammatory stimulus, eliciting local skeletal muscle and systemic immune responses. This chapter presents a standardized physical exercise protocol in mice and subsequent tissue collection, combined with intravital microscopy of skeletal muscle microcirculation and in vitro neutrophil migration assays, to quantify leukocyte recruitment and activation as a versatile model of exercise-induced inflammation.