Objective: Newborn resuscitation guidelines recommend initial assessment of heart rate (HR) and initiation of positive pressure ventilation (PPV) within 60 s after birth in non-breathing newborns. Pulse oximeter (PO) and electrocardiogram (ECG) are suggested methods for continuous HR monitoring during resuscitation. Our aim was to evaluate compliance with guidelines and the efficacy of PO versus ECG monitoring in real-life newborn resuscitations. Methods: In this prospective observational study, we video recorded resuscitations of newborns ≥34 weeks of gestation receiving PPV at birth. Results: 104 resuscitations were analysed. Median (IQR) time from birth to arrival at the resuscitation bay was 48 (22–68) s (n = 62), to initial HR assessment 70 (47–118) s (n = 61), and to initiation of PPV 78 (42–118) s (n = 62). Initial HR assessment (stethoscope or palpation) and initiation of PPV were achieved within 60 s for 35% of the resuscitated newborns. Time to initial HR assessment and initiating PPV was significantly longer following vaginal deliveries than caesarean sections: 84 (70–139) versus 44 (30–66) s (p < 0.001) and 93 (73–139) versus 38 (30–66) s (p < 0.001). Time from birth and sensor application to provision of a reliable HR signal from PO versus ECG was 348 (217–524) (n = 42) versus 174 (105–277) s (n = 30) (p < 0.001) and 199 (77–352) (n = 65) versus 16 (11–22) s (n = 52) (p < 0.001). Conclusion: Initial HR assessment and initiation of PPV were achieved within 60 s after birth in only 1/3 of newborn resuscitations. When applied for continuous HR monitoring, ECG was superior to PO in time to achieve reliable HR signals in real-life resuscitations.