Enhancement of optical resolution in three-dimensional refractive-index tomograms of biological samples by employing micromirror-embedded coverslips

Abstract

Optical diffraction tomography (ODT) enables the reconstruction of the three-dimensional (3D) refractive-index (RI) distribution of a biological cell, which provides invaluable information for cellular and subcellular structures in a non-invasive manner. However, ODT suffers from an inferior axial resolution, due to the limited accessible angles imposed by the numerical aperture of the objective lens. In this study, we propose and experimentally demonstrate an approach to enhance the 3D reconstruction performance in ODT. By employing trapezoidal micromirrors, side scattered signals from the sample are measured for various side plane-wave-illumination angles. By combining the side scattered fields with the forward scattered fields, the axial resolution and 3D image quality of ODT are improved, without changing optical instruments. The feasibility and applicability of the proposed method are demonstrated by reconstructing the 3D RI distribution of a red blood cell and HeLa cells in hydrogel. We also present systematic analyses of the improved 3D imaging performance using numerical simulations and experimental measurements for the 3D transfer function, a point object, and a microsphere. The analyses demonstrate an improved axial resolution of 0.31 μm, 4.8 times smaller than that of the conventional method. The proposed method enables the non-invasive and accurate 3D imaging of 3D cultured cells, which is crucial for cell biology studies.