The purpose of the current research was to develop an instrument for high school students aiming to explore the satisfaction and frustration of their basic psychological needs of autonomy, relatedness, and competence while learning science and to investigate their relation to self-efficacy in science learning. This research involves two studies. In study 1, a total of 392 students (32% females) were solicited from high schools in Taiwan. The results of exploratory factor analysis showed, as expected, that the students' responses were grouped into autonomy satisfaction, autonomy frustration, relatedness satisfaction, relatedness frustration, competence satisfaction, and competence frustration. The regression analysis results indicated that students whose psychological needs were satisfied displayed higher self-efficacy while learning science; however, students whose psychological needs were frustrated showed lower science learning self-efficacy. Study 2 involved 699 students (33.5% females) from high schools in Taiwan. The results of confirmatory factor analysis suggest that the survey items have good construct validity. The results of structural equation modeling also revealed that satisfaction of psychological needs was the positive factor in explaining self-efficacy; on the other hand, frustration of psychological needs did not predict self-efficacy. The findings revealed that the satisfaction of psychological needs of autonomy, relatedness, and competence provides potential impetus for positive functioning with respect to science learning.