Reflection-mode optical diffraction tomography for label-free imaging of thick biological specimens

Optical diffraction tomography (ODT) has emerged as a powerful label-free three-dimensional (3D) bioimaging techniques for observing living cells and thin tissue layers. We report a new reflection-mode ODT (rODT) method for imaging thick biological specimens with ~ 500 nm lateral resolution and ~1 μm axial resolution. In rODT, multiple scattering background is rejected through spatio-temporal gating provided by dynamic speckle-field interferometry, while depth-resolved refractive index maps are reconstructed by developing a comprehensive inverse scattering model that also considers specimeninduced aberration. Benefiting from the high-resolution and full-field quantitative imaging capabilities of rODT, we succeeded in imaging red blood cells and quantifying their membrane fluctuations behind a turbid sample with a thickness of 2.8 scattering mean-free-paths. We further realized volumetric imaging of cornea inside an ex vivo rat eye and quantified its optical properties, including mapping the topography of Dua’s and Descemet’s membrane surfaces on the nanometer scale.

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