Incoherent lateral shearing digital holographic microscopy

Revealing the structural and functional organization of biological samples at submicrometer scales requires high-resolution imaging techniques that provide real-time, quantitative information. However, resolving and analyzing unlabeled biological structures at the subcellular level from a single-shot image remain challenging. We introduce a novel incoherent quantitative phase imaging technique based on lateral shearing digital holographic microscopy, with shearing distances exceeding the coherence area of the employed illumination—a regime not previously achieved and exploited. This strategy consequently ensures artifact-free, high-accuracy, and high-resolution quantitative phase reconstructions. This regime extends single-shot lateral shearing digital holographic microscopy toward incoherent illumination, thus increasing the space–time bandwidth product of the method. The practical common-path geometry also provides enhanced vibration resistance, which is essential for consistent time-lapse measurements. We verify the scalability and effectiveness of the method by investigating samples via multiple condenser–objective pairs, providing a wide range of lateral resolutions and fields of view, thereby ensuring its applicability for various microscope settings. We experimentally verify long-term phase stability and unprecedented phase-reconstruction accuracy. Finally, we use our method to investigate biological samples, including cheek cells, diatoms, and yeast cells, highlighting its potential for dynamic label-free analysis at the organelle level. These results establish our approach as a powerful tool for quantitative phase imaging, enabling high-precision investigations across a wide range of scientific applications.