A 14-year-old, male castrated, Domestic Short Hair cat was presented for thoracic duct ligation via thoracoscopy. To optimise surgical visualisation, the surgeons requested one-lung ventilation. Because adequately sized bronchial blockers or fibreoptic bronchoscopes were not available, a modified single-lumen endotracheal tube guided by a novel method was used. The distance between the incisors, the carina and the mainstem bronchi was premeasured on a computed tomography scan. Once under general anaesthesia, one-lung ventilation was achieved by introducing a guidewire through a modified, previously extended endotracheal tube, confirming its advancement towards the left main bronchus with fluoroscopy, and subsequently advancing the endotracheal tube over the guidewire. Confirmation of one-lung ventilation was initially obtained by observation of unilateral thoracic excursions and later confirmed by thoracoscopic visualisation of right lung atelectasis. The manoeuvre was successfully completed in 8 minutes and no clinically significant complications occurred. Adequate ventilation was achieved using pressure control mode and positive end-expiratory pressure, and allowing permissive hypercapnia. After thoracic duct ligation, bilateral lung ventilation was resumed by slight withdrawal of the endotracheal tube. The cat was allowed to recover from anaesthesia and transferred to the intensive care unit. This report describes a novel method to achieve one-lung ventilation in a cat that may constitute a valid alternative when adequately sized bronchial blockers or fibreoptic bronchoscopes are not available.