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Heartfelt positive emotions, such as love, appreciation, care and compassion, have long been associated with spiritual experience. However, because of a fundamental lack of mental and emotional self-management, such emotions, and associated experiences of increased spiritual connectedness, remain largely transient and unpredictable events in most people’s lives. Here, we summarize our research that has linked sustained positive emotion to a distinct mode of physiological functioning, termed psychophysiological coherence. This mode, characterized by heart rhythm coherence, increased heart-brain synchronization and entrainment of diverse physiological oscillatory systems, is associated with increased emotional stability, improved cognitive performance, and a range of positive health-related outcomes. Additionally, individuals frequently report feelings of increased spiritual connectedness during psychophysiologically coherent states. Using heart rhythm coherence feedback training, individuals can readily learn to self-generate the coherent mode and sustain genuine positive emotional states at will, thus establishing an internal environment that is conducive to fostering spiritual experience.