Scientific thinking constitutes a vital component of scientific competencies, crucial for citizens to adapt to the evolving societal landscape. To cultivate students’ scientific thinking, teachers should possess an adequate professional knowledge foundation, which encompasses pedagogical content knowledge (PCK). Assessing teachers’ PCK of scientific thinking facilitates the development of effective curricula tailored to their continuous professional development. Despite its significance, empirical studies on biology teachers’ PCK of scientific thinking are notably lacking. Hence, this research aimed to create a reliable and valid tool to evaluate upper-secondary school biology teachers’ PCK of scientific thinking. The results showed that the instrument exhibits high reliability and good validity, affirming its efficiency for investigative purposes. A collective of 292 in-service biology teachers from upper-secondary schools participated in this investigation through the completion of an online survey. The results indicated that, overall, as well as for each component, upper-secondary school biology teachers’ performance on PCK of scientific thinking fell within the lower to middle range. Specifically, the performance levels of four components: knowledge of students (KSU), knowledge of instructional strategies (KIS), knowledge of curriculum (KC), and knowledge of assessment (KA) declined sequentially. Keywords: assessment instrument, pedagogical content knowledge, scientific thinking, upper-secondary school biology teacher
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