In science education, learners’ conceptions of scientists and their work are often assessed by the Draw-A-Scientist Test (DAST). Due to validity concerns, methodical literature demands the development of alternative instruments to measure learners’ conceptions validly and efficiently. This study presents an instrument with 29 rating scale items to assess pre-service teachers’ (PSTs) Views of Scientists, their Activities, and Locations (VoSAL). The items were developed based on theoretical considerations, previous findings, and repeated discussions by biology education experts. After several steps of test development, PSTs filled out the questionnaire (N = 1,098). Exploratory factor analyses and reliability measurements mostly confirm the proposed structure. Groups comparisons were performed regarding the results from pre-service biology teachers of three different study stages (nfreshmen = 114; nsecond and third years = 124; ngraduates = 107). Analyses of variance and corresponding post-hoc tests showed that undergraduates (freshmen, second and third years) differ significantly from graduates regarding the scales stereotypical appearance, inquiry location, and scientific activity, with undergraduates having more stereotypical conceptions than graduates. In sum, the VoSAL can be utilized to gain valid data of PSTs’ conceptions about scientists and their work. Also, the VoSAL can be considered efficient since the test time is between 5 and 10 min. Thus, the questionnaire is valuable in studies that aim to introduce and expose PSTs to realistic science images.