Our research is centered on the development of advanced microdroplet-based mass spectrometric methodologies and imaging techniques for real-time, in situ investigation of chemical and biological transformations. By leveraging the unique physicochemical properties of microdroplet surface (e.g., air-water interface), we systematically explore reaction pathways, stabilize transient reactive intermediates, and facilitate environmentally benign synthetic processes. Our contributions have significantly advanced the rapidly evolving field of microdroplet chemistry and mass spectrometry imaging (in biomedical applications such as disease biomarker discovery). Our interdisciplinary research integrates principles from organic and physical chemistry, analytical science, chemical biology, and instrumental development to address key challenges in the elucidation of chemical and biochemical pathways, sustainable reaction engineering, enzyme catalysis, environmental science, and clinical analysis.