INTRODUCTION: Different heel-rise test (HRT) protocols have been used, possibly leading to varied responses. It is necessary to analyse the impact of protocol variation on test responses. PURPOSE: To compare the performance, muscle oxygenation (MO), and heart rate (HR) responses of adults in bilateral HRT protocols. METHODS: This was a cross-sectional crossover study.Thirty participants (23.1±2.9 years; 16 men) performed four bilateral HRT protocols with varying cadence (self-cadenced; externally cadenced) and ankle position (neutral; dorsiflexion). For MO responses, we analysed tissue oxygen saturation (StO2) and oxyhemoglobin concentration variation (∆[O2Hb]) and calculated the variation between the smallest and final values (∆Nadir-Final) and the area under the curve (AUC). The variation between the initial and final HR values (∆HR) and the time constant (τ) were calculated. Friedman’s test was used to compare the variables among the protocols. Two-way ANOVA was used to identify the impact of cadence and/or ankle position. RESULTS: The number of repetitions and execution time were higher in the neutral position and externally cadenced protocols (ptextless0.001 for both). ∆Nadir-Final (StO2: ptextless0.001; ∆[O2Hb]: p=0.005) and AUC (StO2: ptextless0.001; ∆[O2Hb]: ptextless0.001) of both MO variables were higher in the neutral position protocols. Self-cadenced protocols presented higher HR increase and faster τ (p=0.006 and p=0.046). CONCLUSION: Bilateral HRT performed in a neutral position, and external cadence promotes more repetitions and a longer execution time. Dorsiflexion promotes lower muscle reperfusion, and self-cadence higher and faster HR increase.