Objective To evaluate systolic flow-sensitive alternating inversion recovery (FAIR) during rest and exercise stress using 2RR (two cardiac cycles) or 1RR intervals between inversion pulse and imaging. Materials and methods 1RR and 2RR FAIR was implemented on a 3T scanner. Ten healthy subjects were scanned during rest and stress. Stress was performed using an in-bore ergometer. Heart rate, mean myocardial blood flow (MBF) and temporal signal-to-noise ratio (TSNR) were compared using paired t tests. Results Mean heart rate during stress was higher than rest for 1RR FAIR (85.8 +/- 13.7 bpm vs 63.3 +/- 11.1 bpm; p < 0.01) and 2RR FAIR (83.8 +/- 14.2 bpm vs 63.1 +/- 10.6 bpm; p < 0.01). Mean stress MBF was higher than rest for 1RR FAIR (2.97 +/- 0.76 ml/g/min vs 1.43 +/- 0.6 ml/g/min; p < 0.01) and 2RR FAIR (2.8 +/- 0.96 ml/g/min vs 1.22 +/- 0.59 ml/g/min; p < 0.01). Resting mean MBF was higher for 1RR FAIR than 2RR FAIR (p < 0.05), but not during stress. TSNR was lower for stress compared to rest for 1RR FAIR (4.52 +/- 2.54 vs 10.12 +/- 3.69; p < 0.01) and 2RR FAIR (7.36 +/- 3.78 vs 12.41 +/- 5.12; p < 0.01). 2RR FAIR TSNR was higher than 1RR FAIR for rest (p < 0.05) and stress (p < 0.001). Discussion We have demonstrated feasibility of systolic FAIR in rest and exercise stress. 2RR delay systolic FAIR enables non-contrast perfusion assessment during stress with relatively high TSNR.