This article describes the development and validation process of a scale designed to assess the assistive technology (AT) knowledge of preservice special education teachers related to visual impairment (VI). The scale was administered to two groups of preservice special education teachers, and data from the first group (N = 291) were subjected to an exploratory factor analysis (EFA). The results of the EFA revealed that the scale had a five-factor structure, which included "braille technologies," "accessibility for documents and software," "AT for diverse needs," "AT for accessing print," and "AT for transforming content into accessible digital format." Based on the findings of the EFA, the researchers revised the scale and applied it to the second group of preservice teachers (N = 119) to confirm the five-factor structure through a confirmatory factor analysis (CFA). The fit indices calculated during the CFA indicated a good model fit. By following a systematic approach, a valid 23-item scale for assessing AT knowledge related to VI was developed. This study offers a useful assessment tool and knowledge constructs for designing AT courses. A newly developed, valid, and reliable measurement tool has been created to gauge the self-efficacy levels of preservice special education teachers with respect to specific knowledge related to AT for VI. Both practitioners and researchers can employ this developed scale to assess the self-efficacy of preservice teachers in their knowledge and application of AT for VI, as well as to evaluate the effectiveness of AT courses. The knowledge constructs that have been identified in this study, based on the results of EFA and CFA, can serve as valuable guidance for instructors when designing the content of courses related to AT.