Background The development of a mobile telephone food record has the potential to ameliorate much of the burden associated with current methods of dietary assessment. When using the mobile telephone food record, respondents capture an image of their foods and beverages before and after eating. Methods of image analysis and volume estimation allow for automatic identification and volume estimation of foods. To obtain a suitable image, all foods and beverages and a fiducial marker must be included in the image. Objective To evaluate a defined set of skills among adolescents and adults when using the mobile telephone food record to capture images and to compare the perceptions and preferences between adults and adolescents regarding their use of the mobile telephone food record. Methods We recruited 135 volunteers (78 adolescents, 57 adults) to use the mobile telephone food record for one or two meals under controlled conditions. Volunteers received instruction for using the mobile telephone food record prior to their first meal, captured images of foods and beverages before and after eating, and participated in a feedback session. We used chi-square for comparisons of the set of skills, preferences, and perceptions between the adults and adolescents, and McNemar test for comparisons within the adolescents and adults. Results Adults were more likely than adolescents to include all foods and beverages in the before and after images, but both age groups had difficulty including the entire fiducial marker. Compared with adolescents, significantly more adults had to capture more than one image before (38% vs 58%, P = .03) and after (25% vs 50%, P = .008) meal session 1 to obtain a suitable image. Despite being less efficient when using the mobile telephone food record, adults were more likely than adolescents to perceive remembering to capture images as easy (P < .001). Conclusions A majority of both age groups were able to follow the defined set of skills; however, adults were less efficient when using the mobile telephone food record. Additional interactive training will likely be necessary for all users to provide extra practice in capturing images before entering a free-living situation. These results will inform age-specific development of the mobile telephone food record that may translate to a more accurate method of dietary assessment.