Metabolic stress is a primary mechanism of muscle hypertrophy and is associated with microvascular oxygenation and muscle activation. Considering that drop-set (DS) and crescent pyramid (CP) resistance training systems are recommended to modulate these mechanisms related to muscle hypertrophy, we aimed to investigate if these resistance training systems produce a different microvascular oxygenation status and muscle activation from those observed in traditional resistance training (TRAD). Twelve volunteers had their legs randomized in an intra-subject cross-over design in TRAD (3 sets of 10 repetitions at 75% 1-RM), DS (3 sets of ∼50-75% 1-RM) and CP (3 sets of 6-10 repetitions at 75-85% 1-RM). Vastus medialis microvascular oxygenation and muscle activation were respectively assessed by non-invasive near-infrared spectroscopy and surface electromyography techniques during the resistance training sessions in the leg-extension exercise. Total hemoglobin area under the curve (AUC) (TRAD: -1653.5 ± 2866.5; DS: -3069.2 ± 3429.4; CP: -1196.6 ± 2675.3) and tissue oxygen saturation (TRAD: 19283.1 ± 6698.0; DS: 23995.5 ± 15604.9; CP: 16109.1 ± 8553.1) increased without differences between protocols (p>0.05). Greater decreases in oxygenated hemoglobin AUC and hemoglobin differentiated AUC were respectively found for DS (-4036.8 ± 2698.1; -5004.4 ± 2722.9) compared with TRAD (-1951.8 ± 1720.0; -2250.3 ± 1305.7) and CP (-1814.4 ± 2634.3; 2432.2 ± 2891.4) (p<0.03). Higher increases of hemoglobin deoxygenated AUC were found for DS (1426.7 ± 1320.7) compared with TRAD (316.0 ± 1164.9) only (p=0.04). No differences were demonstrated in electromyographic amplitudes between TRAD (69.0 ± 34.4), DS (61.3 ± 26.7) and CP (60.9 ± 38.8) (p>0.05). Despite DS produced lower microvascular oxygenation levels compared with TRAD and CP, all protocols produced similar muscle activation levels.