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This study aimed to develop and validate the Motivated Strategies for Science Learning (MSSL) instrument, which measures motivation and strategy utilization in elementary students' science learning. The MSSL was administered to 587 third-grade students in rural Texas schools following a literacy-infused science intervention. Psychometric analysis, including exploratory and confirmatory factor analyses, yielded satisfactory results for the instrument's properties. Additionally, measurement invariance analyses confirmed the MSSL's reliability and validity across online and in-person groups, affirming its effectiveness in accurately measuring students' science motivation. The validated MSSL provides valuable insights for science education research, offering educators a tool to measure students' science learning motivation in diverse learning environments.