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Objective: The purpose of this study is to understand the nature and source of disruptions in an ambulance during the telemedicine-based caregiving process for stroke patients to enhance the ambulance design for supporting telemedicine-based care. Background: Telemedicine is emerging as an efficient approach to provide timely remote assessment of patients experiencing acute stroke in an ambulance. These consults are facilitated by connecting the patient and paramedic with a remotely located neurologist and nurse using cameras, audio systems, and computers. However, ambulances are typically retrofitted to support telemedicine-enabled care, and the placement of these systems inside the ambulance might lead to spatial challenges and disruptions during patient evaluation. Method: Video recordings of 13 simulated telemedicine-based stroke consults were coded and analyzed using an existing systems-based flow disruption (FD) taxonomy. For each observed disruption—the type, severity or impact, location in the ambulance, and equipment involved in the disruption were recorded. Results: Seat size, arrangement of assessment equipment, location of telemedicine equipment (computer workstation), and design of telemedicine camera were among the factors that impacted telemedicine-related disruptions. The left ambulance seat zone and head of the patient bed were more involved in environmental hazard–related disruptions, while the right zone of the ambulance was more prone to interruptions and communication-related disruptions. Conclusion: Adequate evaluation space for the paramedic, proper placement of evaluation equipment, and telemedicine computer location could facilitate the stroke care evaluation process and reduce FDs in the ambulance.