This study explores the color preferences of elderly residents of nursing homes based on the expected activities for different rooms, activity rooms, and bedrooms and compares the results with the arousal level induced by each color. Two experiments were conducted, one with elderly people living in nursing homes in the Valencian Community, and the other in a laboratory with virtual reality and physiological markers (i.e., heart rate variability). Individuals assessed six colors in two groups of warm and cool. Results demonstrate that preferences for warm and cool colors depend on the room type. For the activity room, warm colors were preferred over the cool colors by both genders, fitting the higher arousal levels induced by warm versus cool colors. For the bedrooms, cool colors were preferred by both genders, fitting the lower arousal levels induced by cool colors in females and the other models that suggest a U-shaped relation between the arousal level and the visible spectra of colors. Therefore, the color preferences for interior spaces in nursing homes depend on the room type and are related to the arousal level for the expected activity in them.