Background: Ischemic stroke is the leading cause of long-term disability in the United States. To reduce the likelihood of debilitating outcomes, early treatment with fibrinolytics is necessary; however, many struggle to identify strategies that limit time to treatment. The purpose of this study is to identify if implementing a process for obtaining laboratory blood draws by emergency medical services (EMS) prior to patient arrival at the hospital will reduce the door-to-needle time for patients arriving with acute ischemic stroke. Objectives: To compare the door-to-needle time for patients with acute ischemic stroke who had blood drawn via EMS versus in hospital. Methods: This single-center, retrospective cohort study was conducted from 2015 to 2017 at a 433-bed designated Level II Trauma Center and Comprehensive Stroke Center community hospital. A comprehensive chart review was done via stroke coordinator binders and electronic medical record. Adult patients ≥18 years of age with an acute ischemic stroke diagnosis who met criteria for and received alteplase were included in the study. Patients were excluded if they did not arrive via EMS. The primary end point assessed time to fibrinolytic treatment via door-to-needle times. Results: A total of 66 patients were included with 28 in the EMS laboratory drawn group and 38 in the usual group. Median door-to-needle time was 57.5 ± 19.0 minutes in the EMS group and 55.0 ± 31.9 minutes in the usual care group (p = 0.818). Conclusions: Collection of blood for laboratories by EMS personnel did not significantly shorten door-to-needle time.