Background: Hope can affect the thinking habits, emotional regulations, and behaviors of individuals. Hope is considered as a positive trait by clinicians, who often assess the level of hope in psychological evaluations. Previous measurements of hope were largely based on self-reported questionnaires leading to the problem of subjectivity. Heart Rate Variability (HRV) is a bio index that is an objective, quick, cost effective, and non-invasive measurement. HRV has been used in the evaluation of physical health and some psychiatric conditions. However, it has not been tested for its potential to be a bio-index of the level of hope. Method: This pilot cross-sectional observational study aimed to examine the relationships between HRV and the level of hope among adult Chinese people in Hong Kong. Convenience sampling was used and 97 healthy participants were recruited. Their level of hope was measured by the Dispositional Hope Scale-Chinese (DHS-C), and their HRV was quantified by emWave Pro Plus, a reliable sensor of HRV. Spearman’s correlation coefficient analysis was performed on the HRV measurements and DHS-C. Results: The DHS-C’s overall mean score was 45.49. The mean scores of the subscale DHS-C (Agency) was 22.46, and the mean scores of DHS-C (Pathway) was 23.03. It was also revealed that there were significant, weak, and negative correlations between the level of hope and four out of ten HRV metrics. One HRV metric was found to have a significant, weak, and positive correlation with the level of hope. Conclusion: This study provided initial evidence to support the use of HRV as a bio-index of hope. Implications of the current study and recommendations for future research directions are discussed.