Enabling Super-Resolution Quantitative Phase Imaging via OpenSRQPI—A Standardized Plug-and-Play Open-Source Tool for Digital Holographic Microscopy with Structured and Oblique Illumination

Accurate and label-free quantitative phase imaging (QPI) plays a crucial role in advancing diagnostic techniques that streamline histology and diagnostic procedures by minimizing sample preparation time, resources, and requirements. Although Digital Holographic Microscopy (DHM) has become a prominent tool within QPI, its diffraction-limited resolution has hindered broader adoption of QPI-DHM. The use of structured and oblique illumination in DHM platforms has overcome the resolution limit, advancing QPI-DHM technology to super-resolution QPI. Despite demonstrated success, adoption of super-resolution DHM (SR-DHM) in clinical and biomedical research remains limited by the absence of a standardized reconstruction algorithm capable of delivering quantitatively accurate, distortion-free super-resolved phase images. This work presents OpenSRQPI, the first standardized computational framework for super-resolution phase reconstruction in DHM systems, whether using structured or oblique illumination. Through its intuitive graphical user interface (GUI) and minimal parameter requirements, OpenSRQPI reduces the technical barrier for non-experts, making super-resolution QPI broadly accessible, enabling new studies of live-cell dynamics, subcellular structure, and tissue morphology.