We report the implementation of an interferometric null test using a high-definition spatial light modulator (SLM) as a reconfigurable alternative to a computer-generated hologram. We detail the alignment process chain, including novel techniques using the SLM to project alignment fiducials on the test part. To validate the alignment protocol, we measure a mild off-axis conic with the SLM-based system and cross-validate with conventional interferometry within 30 nm root-mean-square (RMS) surface figure. Finally, we report the null test of a 65 mm clear aperture concave freeform with 91 μm peak-valley sag departure from the base sphere. The measured surface figure of the freeform is within 40 nm RMS compared to the measurement with a commercial metrology instrument.

A vibration-insensitive, single-shot phase-calibration method for phase-only spatial light modulators (SLM) is reported. The proposed technique uses a geometric phase lens to form a phase-shifting radial shearing interferometer to enable common-path measurements. This configuration has several advantages: (a) unlike diffraction-based SLM calibration techniques, this technique is robust against intensity errors due to misalignment; (b) unlike two-beam interferometers, this technique offers a high environmental stability; and (c) unlike intensity-based methods, the phase-shifting capability provides a phase uncertainty routinely in the order of 2 π / 100 . The experimental results show a significantly higher accuracy when compared to the diffraction-based approaches.

A quantitative phase measuring technique is presented that estimates the object phase from a series of phase shifted interferograms that are obtained in a common-path configuration with unknown phase shifts. The derived random phase shifting algorithm for common-path interferometers is based on the Generalized Phase Contrast theory [pl. Opt. 40(2), 268 (2001) [CrossRef] ], which accounts for the particular image formation and includes effects that are not present in two-beam interferometry. It is shown experimentally that this technique can be used within common-path configurations employing nonlinear liquid crystal materials as self-induced phase filters for quantitative phase imaging without the need of phase shift calibrations. The advantages of such liquid crystal elements compared to spatial light modulator based solutions are given by the cost-effectiveness, self-alignment, and the generation of diminutive dimensions of the phase filter size, giving unique performance advantages.

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.

Kudsi R, et al. J Periodontal Implant Sci. 2024 Dec;55:e5. https://doi.org/10.5051/jpis.2401320066