Abstract
•Vortex-Legendre method effectively compensates phase aberrations in digital holographic microscopy for complex biological samples without iterative steps or additional optics.•The method employs numerical optical vortices for tilt aberration correction, enhancing subsequent Legendre polynomial fitting for high-order aberrations.•Experimental validation demonstrates preservation of high-frequency spatial details across both telecentric and non-telecentric configurations, crucial for intricate biological samples.•Benchmarking reveals this method outperforms existing approaches in quantitative accuracy and computational efficiency, proving suitable for biomedical imaging applications.
Digital holographic microscopy (DHM) offers label-free, high-resolution quantitative phase imaging, making it a powerful tool for real-time visualization of dynamic biological processes. However, when imaging intricate biological samples with detailed cellular structures—such as tissues containing diverse cell types, fine organelles, or intricate vascular networks— and with subtle variations in refractive index, the accuracy of phase reconstruction is compromised by several types of phase aberrations. These aberrations include tilt distortions due to the off-axis configuration, quadratic phase errors introduced by microscope objectives, and additional higher-order aberrations caused by sample heterogeneity and imperfections in the optical system. Traditional compensation techniques either rely on iterative computations, multi-shot acquisitions, or additional optical components, thereby limiting their applicability in fast, real-time imaging scenarios. This work introduces a novel, fully computational hybrid approach—termed the vortex-Legendre method—that addresses these limitations. This method leverages a numerical optical vortex to achieve precise, sub-pixel localization of the +1 diffraction order for tilt aberration correction, followed by Legendre polynomial fitting to efficiently compensate for residual higher-order aberrations. Validation experiments on calibrated phase targets and various biological samples demonstrate that the vortex-Legendre method preserves high-frequency details and delivers consistent performance across both telecentric and non-telecentric imaging configurations. Compared to state-of-the-art approaches, this method improves phase compensation accuracy while maintaining computational efficiency, paving the way for high-fidelity quantitative phase imaging of complex biomedical samples.