PROJECT SUMMARY / ABSTRACT The overarching research goal of the PI’s research program aims to advance imaging science and technology to transform biomedical research. To date, a complete understanding is still lacking to elucidate how biomole- cules, organelles, and microenvironments are assembled within single cells, their variations over large popula- tions, and their integrated roles in cell and tissue functions, developments, and disease initiation and therapeutics. There have been major unmet imaging needs to probe the intracellular and multi-parametric complexities and heterogeneity in cells and tissues with 3D nanoscale resolution, volumetric capability, high throughput and sen- sitivity, and platform generalizability. To address the demands, the PI’s laboratory investigates the novel physical, engineering, and instrumental principles and systems and deploys them to illuminate both fundamental and medical discoveries. This renewal proposal aims to continue the efforts and establish and strengthen the pro- gram’s leadership in systems biophotonics at the critical interface between imaging innovations and life sciences. Specifically, the PI proposes the research program to proceed in three major directions to provide enabling technologies that overcome imaging challenges in space, time, and accessibility for a deeper understanding of biological complexities: (1) wave physics and super-resolution microscopy to probe intracellular complexities and heterogeneity with 3D ultrahigh-resolution, volumetric capability, high throughput and sensitivity, and platform generalizability; (2) light-field microscopy and computational microscopy to enable the interrogation and ultrafast, in vivo imaging of multi-scale, volumetric biological dynamics and activities; (3) miniature microscopy and camera physics to transform conventional imaging platforms and enhance imaging device accessibility to wide-ranging imaging conditions, modalities, and biological systems. The successful accomplishment and dissemination of the proposed research are anticipated to (i) promise and catalyze the discovery and translation of imaging sci- ence and technology, (ii) provide methodological avenues and revolutionize biomedical investigations restrained by conventional methods, and (iii) transform existing imaging infrastructure, laying a critical intellectual founda- tion for broader science, engineering, and technology breakthroughs.