Improved characterization of arrhythmias is based on minimally invasive catheterizations. However, these catheterizations have been poorly explored in horses because apart from 3-dimensional (3D) mapping systems, continuous guidance of the catheter's position with adequate detail is difficult using current imaging modalities. Position multiple electrophysiology catheters simultaneously at predetermined strategical positions in the heart using transthoracic echocardiographic guidance. Eight adult healthy horses. Observational study. Two electrophysiological studies were performed: 1 procedure with catheters positioned in the right heart in the standing sedated horse and 1 procedure under general anesthesia with catheters positioned in the left heart. Except for the coronary sinus catheter, each catheter positioning was simultaneously guided by right-parasternal transthoracic echocardiography and 3D electro-anatomical mapping. For each catheter position, a central imaging plane was taken as the starting point, after which the imaging probe was shifted, rotated, and angulated to visualize the catheter over its entire length, including its distal electrode. Catheter positionings in the right heart and left ventricle were successfully guided in the majority of the horses whereas catheter positionings in the left atrium, and especially the pulmonary veins, were challenging to guide echocardiographically. Ultrasound guidance of catheters to specific positions useful for electrophysiological mapping was feasible in the right heart and left ventricle but challenging for the left atrium. This approach creates a perspective for minimally invasive arrhythmia diagnosis without the need for a 3D mapping system. Left parasternal views and intracardiac echocardiography might provide better guidance for left atrial positions.