Context: Several interventions are available to reduce the intensity and duration of the unwanted effects (eg, muscle soreness) associated with physical activity, such as massage, compression garments, and sequential pulse compression (SPC). Such interventions aim to increase blood flow to alleviate symptoms. However, there is a lack of evidence to support the use of SPC to alter total hemoglobin concentration (THb) in active individuals. Objective: To examine the acute effects of a single session of SPC on hemoglobin concentration compared with a control condition. Design: Single cohort, crossover design. Participants: Thirty-four physically active and healthy participants (females = 12 and males = 22) completed the study. Interventions: The authors randomly assigned participants to first receive the experimental (SPC) or control condition. Measures were recorded precondition and postcondition. Participants returned to the laboratory to complete the second condition ≥24 hours after the first condition. Main Outcome Measures: Relative changes in THb, deoxygenated hemoglobin, and oxygenated hemoglobin measures were recorded using near-infrared spectroscopy placed on the muscle belly of the medial gastrocnemius of the dominant limb. Results: SPC significantly increased THb (P < .001, d = 0.505) and oxygenated hemoglobin (P < .001, d = 0.745) change scores compared with the control condition. No statistical difference in deoxygenated hemoglobin change scores was found between the SPC and control conditions, but a medium effect size suggests potential biological significance (P = .06, d = 0.339). Conclusions: Overall, SPC increases THb to the lower-extremity and may be a viable option in the management of muscle soreness related to physical activity.