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Abstract There is a significant gap in cost-effective quantitative phase microscopy (QPM) systems for studying dynamic cellular processes while maintaining accuracy for long-term cellular monitoring. Current QPM systems often rely on complex and expensive voltage-controllable components like Spatial Light Modulators or two-beam interferometry. To address this, we introduce a QPM system optimized for time-varying phase samples using azobenzene liquid crystal as a Zernike filter with a polarization-sensing camera. This system operates without input voltage or moving components, reducing complexity and cost. Optimized for gentle illumination to minimize phototoxicity, it achieves a 1 Hz frame rate for prolonged monitoring. The system demonstrated accuracy with a maximum standard deviation of ±42 nm and low noise fluctuations of ±2.5 nm. Designed for simplicity and single-shot operations, our QPM system is efficient, robust, and precisely calibrated for reliable measurements. Using inexpensive optical components, it offers an economical solution for long-term, noninvasive biological monitoring and research applications. 

This study showcases the multifunctionality of a single-shot quantitative phase microscopy (QPM) system for comprehensive cell analysis. The system captures four high-contrast images in one shot, enabling tasks like cell segmentation, measuring cell confluence, and estimating cell mass. We demonstrate the usability of the QPM system in routine biological workflows, showing how its integration with computational algorithms enables automated, precise analysis, achieving accuracy scores between 85% and 97% across samples with varying cell densities, even those with low signal-to-noise ratios. This cost-effective tool operates under low-intensity light and resists vibrations, making it highly versatile for researchers in both optical and biological fields.