The aim in this study was to understand current practice and use of nitrous oxide for management of procedural-related pain and procedural anxiety, to identify perceived barriers to use of nitrous oxide and to develop an understanding of patients, families, and nurse awareness and knowledge of the use of nitrous oxide in an Australian tertiary pediatric oncology/hematology short stay unit. Three online questionnaires (patients, parents, and nursing staff) were developed and completed between September and November 2015. Most children and young people (61%) report receiving nitrous oxide for at least one procedure. Patients, parents, and nurses rated nitrous oxide as highly effective and would like more access to nitrous oxide for the child’s pain management. Several barriers to use were reported. These findings suggest that nitrous oxide is effective for pain management; however, its use is inconsistent. Findings can potentially develop standardized processes and improve nurse education and accreditation, which may increase the safety, efficacy, and utilization of nitrous oxide for children’s procedural pain management.