<p>Meditation practices, including mindfulness, are linked with adaptive emotional processing and regulation. Although startle response modulation among meditators has been studied using habituation and prepulse-induced startle inhibition paradigms, affective startle modulation, which refers to potentiation by negative stimuli and attenuation by positive stimuli (both relative to neutral stimuli), remains unexplored. This study examined how regular meditation practice, dispositional mindfulness, and affective difficulties influence affective modulation of the acoustic startle reflex. Seventeen meditators and thirty non-meditators were exposed to pleasant, neutral, and unpleasant images while their eye-blink startle responses were recorded. Participants also completed self-report measures of dispositional mindfulness, alexithymia, emotion regulation difficulties, depression, anxiety, and stress. Meditators, compared to non-meditators, reported higher dispositional mindfulness, particularly in the <i>Observing</i> and <i>Non-reactivity</i> domains, lower stress, and fewer difficulties in goal-oriented behaviour during negative emotions; they also had longer startle onset latencies, potentially indicating lower state anxiety, across the entire experiment regardless of the valence of visual images. Higher dispositional mindfulness correlated with lower scores on alexithymia, emotion regulation difficulties, depression, anxiety, and stress across the pooled sample. These findings suggest that mindfulness, whether cultivated through meditation or as a trait, reduces negative emotionality, highlighting its potential for emotional regulation and stress reduction.</p>