Background: Previous studies on adult patients in hospital isolation show that it can be highly stressful, with some patients affected by depression and anxiety. This study aimed to explore how children, adolescents, and their parents perceive isolation and how it affects them physically, psychologically, and socially. Methods: A qualitative approach was used comprising semi-structured interviews, participant observations, and field notes. Findings: Two adolescents and 13 parents were selected to participate in the interviews, while three children and their parents participated in participant observation. Besides the participant observation one of the adolescents and one parent also participated in interviews. The children and adolescents were isolated at the hospital due to bacterial or viral infections or were carriers of an antibiotic resistant bacteria. After performing a thematic analysis to identify patterns in the data, four themes emerged: 1. Differing information provided about bacteria, virus, hygiene precautions, and isolation, 2. Dependence on healthcare professionals, 3. Impact of the environment, and 4. Coping and psychological reactions on isolation. Discussion: The participants conveyed positive and negative experiences and emotions during isolation in relation to daily activities, psychological well-being, and social life. Adolescents and their parents requested clear comprehensible and consistent information on isolation from healthcare professionals. Application to practice: To improve hospital isolation for children, adolescents, and their parents, well-written information on isolation and hygiene precautions is crucial, as are evolving strategies to minimise social exclusion and the emotional impact of isolation.