Background: Rabbits have a longer inter-blink time (approximately 10 minutes) compared with humans (five to eight seconds), suggesting that rabbits have a much more stable tear film. Using fluorescein, the tear break-up time of rabbits has been reported to be similar to that of humans. This study set out to measure the tear break-up time in rabbits using non-invasive methods and to establish the pattern of tear break-up compared to humans. Methods: The tear break-up time was measured and the pattern of tear break-up was observed in six New Zealand White rabbits on two separate occasions using both the Keeler Tearscope-plusTM and a slitlamp biomicroscope. Results: The mean rabbit tear break-up time was 29.8 ± 3.4 (SD) minutes. This contrasts with the reports of human tear break-up time of eight to 30 seconds. The tear breaking spread very slowly and was often restricted to the area of the initial break. Conclusion: Rabbit tears have a significantly higher tear break-up time than humans and this aligns with previously demonstrated differences in inter-blink time between rabbits and humans. Understanding the underlying mechanisms of tear stability may lead to novel ways of increasing human tear film stability.