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Background: The role of stroke nurses in patient selection and administration of recombinant tissue plasminogen activator (rt‐PA) for acute ischaemic stroke is evolving. Objectives: To compare differences in stroke nurses' practices related to rt‐PA administration in Australia and the United Kingdom (UK) and to examine whether these differences influence rt‐PA treatment rates. Methods: A cross‐sectional, self‐administered questionnaire administered to a lead stroke clinician from hospitals known to provide rt‐PA for acute ischaemic stroke. Chi‐square tests were used to analyse between‐country differences in ten pre‐specified rt‐PA practices. Non‐parametric equality of medians test was used to assess within‐country differences for likelihood of undertaking practices and association with rt‐PA treatment rates. Reporting followed STROBE checklist. Results: Response rate 68%; (Australia: 74% [n = 63/85]; UK: 65% [n = 93/144]). There were significant differences between countries for 7/10 practices. UK nurses were more likely to: request CT scan; screen patient for rt‐PA suitability; gain informed consent; use telemedicine to assess, diagnose or treat; assist in the decision for rt‐PA with Emergency Department physician or neurologist; and undergo training in rt‐PA administration. Reported median hospital rt‐PA treatment rates were 12% in the UK and 7.8% in Australia: (7.8%). In Australia, there was an association between higher treatment rates and nurses involvement in 5/10 practices; read and interpret CT scans; screen patient for rt‐PA suitability; gain informed consent; assess suitability for rt‐PA with neurologist/stroke physician; undergo training in rt‐PA administration. There was no relationship between UK treatment rates and likelihood of a stroke nurse to undertake any of the ten rt‐PA practices. Conclusion: Stroke nurses' active role in rt‐PA administration can improve rt‐PA treatment rates. Models of care that broaden stroke nurses' scope of practice to maximise rt‐PA treatment rates for ischaemic stroke patients are needed. Relevance to clinical practice: This study demonstrates that UK and Australian nurses play an important role in thrombolysis practices; however, they are underused. Formalising and extending the role of stroke nurses in rt‐PA administration could potentially increase thrombolysis rates with clinical benefits for patients. [ABSTRACT FROM AUTHOR]