The development of effective measures to understand anxiety in future teachers is essential. Research shows that the initial, training stages of a teacher’s career are often accompanied by heightened anxiety, driven by the challenges of managing academic assessment demands, gaining confidence in classroom management skills, and facing new teaching environments. This study employed a two-phase, mixed-methods approach to develop and pilot a new questionnaire to measure stress and anxiety in future teachers. Phase 1 involved informal observations and interviews to identify markers of anxiety and inform questionnaire design. Phase 2 piloted the questionnaire with participants enrolled in a teacher training course at the University of Palermo, assessing the internal consistency and exploratory factor analysis to guide future item refinement and domain structure. Preliminary results demonstrated high reliability for the total scale (Cronbach’s α = 0.966) and individual subscales addressing Classroom Management and Student Engagement (α = 0.900), Workload and Teaching Stress (α = 0.903), Performance and Evaluation Anxiety (α = 0.895), and Emotional and Physical Manifestations (α = 0.896). Exploratory factor analysis then revealed three distinct factors, suggesting a need for revisions to improve alignment with theoretical constructs. Qualitative participant feedback highlighted opportunities to enhance the questionnaire by addressing areas not yet covered, such as systemic issues like precarious teaching conditions and bureaucratic stress, along with suggestions regarding scale length and item clarity. This initial analysis establishes the questionnaire as a foundation for future refinement and validation, essential for triangulating data with emerging technologies in subsequent research.