Examining the effect of clinical nurses' sleep characteristics on work stress and anxiety can enhance medical care. This study explored the relationships among work stress, anxiety, and sleep characteristics among nurses with sleep disturbance. A purposive sampling design was applied in this survey research. A total of 85 female nurses with sleep disturbance who worked in regional teaching hospitals in New Taipei City for more than 3 months and scored more than 5 points on the Pittsburgh Sleep Quality Index were recruited for this study. Data were collected using structured questionnaires, which measured sleep status, sleep characteristics, work stress, and anxiety. The results of this research showed that participants suffering from sleep disturbance were on average 31.1 years old and had worked for an average of 12.4 years. On average, their time in bed (TIB), total sleep time, and sleep efficiency (SE) were 7.5 hours, 5.9 hours, and 81.8%, respectively. Moreover, 17.1% of nurses had moderate anxiety, whereas 4.1% suffered from severe anxiety. Higher work stress among the nurses with sleep disturbance was significantly associated with less TIB and higher SE; this was a positive correlation. Moreover, anxiety was significantly associated with higher SE, more years of work experience, higher life pressure, lower health status, and higher work stress. Work stress was the anxiety-related factor that could explain the variation of 15.6%, based on regression analysis. The results provide a reference for future research.