The intra-aortic balloon pump (IABP) provides circulatory support through counterpulsation. The hemodynamic effects of the IABP may vary with assisting frequency and depend on IAB inflation/deflation timing. We aimed to assess in vivo the IABP benefits on coronary, aortic, and left ventricular hemodynamics at different assistance frequencies and trigger timings. Six healthy, anesthetized, open-chest sheep received IABP support at 5 timing modes (EC, LC, CC, CE, CL, corresponding to early/late/conventional/conventional/conventional inflation and conventional/conventional/conventional/early/late deflation, respectively) with frequency 1:3 and 1:1. Aortic (Q(ao)) and coronary (Q(cor)) flow, and aortic (P(ao)) and left ventricular (PLV) pressure were recorded simultaneously, with and without IABP support. Integrating systolic Q(ao) yielded stroke volume (SV). EC at 1:1 produced the lowest end-diastolic P(ao) (59.5 ± 7.8 mmHg [EC], 63.4 ± 11.1 mmHg [CC]), CC at 1:1 the lowest systolic PLV (69.1 ± 6.5 mmHg [CC], 76.4 ± 6.5 mmHg [control]), CC at 1:1 the highest SV (88.5 ± 34.4 ml [CC], 76.6 ± 31.9 ml [control]) and CC at 1:3 the highest diastolic Qcor (187.2 ± 25.0 ml/min [CC], 149.9 ± 16.6 ml/min [control]). Diastolic P(ao) augmentation was enhanced by both assistance frequencies alike, and optimal timings were EC for 1:3 (10.4 ± 2.8 mmHg [EC], 6.7 ± 3.8 mmHg [CC]) and CC for 1:1 (10.8 ± 6.7 mmHg [CC], -3.0 ± 3.8 mmHg [control]). In our experiments, neither a single frequency nor a single inflation/deflation timing, including conventional IAB timing, has shown superiority by uniformly benefiting all studied hemodynamic parameters. A choice of optimal frequency and IAB timing might need to be made based on individual patient hemodynamic needs rather than as a generalized protocol.