Background: Stroke telemedicine enables “Hub” hospitals with stroke expertise to interact with smaller “Spoke” hospitals in an effort to improve capabilities of accurate diagnosis and treatment of patients suffering from an acute stroke event. Administration of recombinant tissue plasminogen activator (rtPA) for treatment for acute ischemic stroke (AIS) patients is time dependent, and telemedicine has the potential to increase AIS patient thrombolytic treatment rate. Objective: To compare treatment rates of AIS patients in the Spoke hospital setting before and after implementation of telemedicine. Methods: The study included patients (age≥18) hospitalized with a primary diagnosis of AIS (ICD-9 codes 433.x1, 434.x1, 436) admitted to the hospital between January 1, 2006 and December 30, 2012 from 13 pre-specified Spoke hospitals in the Premier research database with telemedicine implementation ranging from October 2009 through December 2011. Evidence of MRI or CT scan during the hospital stay was required for study inclusion. Treatment of rtPA was identified using ICD-9 procedure code 99.10 and/or charge master data indicating rtPA use. Results: A total of 9,629 and 5,228 patients were included in the study population pre- and post- telemedicine implementation respectively. Mean age and gender distribution were similar for pre- and post- implementation (72.0 vs. 71.3 years; 53.1% vs. 51.9% female, respectively). The percentage of patients treated with rtPA increased from 4.4% during pre- to 6.9% during post-implementation (p <0.0001) time period, a 56.8% increase. Increase in treatment percentages were seen in hospitals with <200 beds (1.1% vs. 7.3% p<0.0001) and hospitals with >= 200 beds (5.2% vs. 6.8% p=0.0003). Conclusions: Results from the study demonstrate that the percent of patients treated with rtPA with telemedicine increased more than 50% as compared to the pre-telemedicine time period. Smaller hospitals showed the most significant increase in rtPA treatment rate. This finding supports previous findings that telemedicine improves appropriate use of thrombolytic treatment for AIS patients and highlights the impact in smaller hospital settings.