Cat caregivers face many potential barriers when accessing veterinary care, such as cat-related challenges with carriers, travel, and the clinic environment, as well as time away from home and/or work, distance to the veterinary clinic, and costs of veterinary care. A potential solution to overcoming some of these barriers may be the use of video telemedicine: using video technology to deliver health and behavior care, education, and information remotely. This study recruited companion cat caregivers and their cats (n = 30) to participate in two mock spay re-check examinations: an in-clinic appointment and a virtual video telemedicine appointment. During the mock examinations, cat behavioral and physiological responses were assessed. Order of appointment was counterbalanced, and caregivers completed a pre- and post- appointment questionnaire to assess attitudes and experience with the in-clinic versus video appointments before and after participation. The study results show that cats had a higher respiratory rate (p < 0.001), larger pupil dilation (p < 0.001), and more negative ear positions (p = 0.015) during the mock clinic appointment compared to the video appointment. Cats were also more likely to leave the handling area after the mock clinic examination compared to the video examination (p = 0.001). In addition, caregiver ratings of the video appointment were very positive (62.1%) or somewhat positive (34.5%), and participants stated it was very easy to access the technology (58.6%), internet (65.5%), and video conferencing platform (75.9%) needed. Post-survey participant ratings indicated that video telemedicine was perceived to be more helpful than in-clinic visits for reducing caregiver stress (p = 0.029) and cat stress (p < 0.0001). There was also a post-survey difference in caregiver ratings on accessibility of veterinary care between video telemedicine and clinic appointments (p < 0.0001), with results indicating that video telemedicine increases caregivers’ access to cat care. Thus, from cat caregiver perspectives, video telemedicine may reduce stress and increase accessibility to cat health care compared to clinic visits. Results also suggest that video appointments reduce negative responses in cats compared to in-clinic visits, and thus may be particularly useful for cats who show higher levels of stress in a clinic environment. More research is needed to evaluate cat responses during other appointment types for further assessment and replication of the study results.