Imaging models for three-dimensional transmitted-light DIC microscopy

Abstract

Nomarski Differential-Interference-Contrast (DIC) microscopy is a widely used method for imaging transparent specimens that are not visible with ordinary light microscopy. DIC microscopy enhances contrast in the images of such specimens by converting differential phase changes to intensity variations via the method of light interference. These phase changes are introduced in light as it passes through regions of different refractive index within a specimen. In this paper, the development of an imaging model that describes 3D DIC imaging under partially-coherent illumination is presented. Our approach in deriving the model involves the derivation of a 2D model and its extension to three dimensions, assuming weak optical interactions within the specimen. The coherent limit of our 2D model coincides with existing DIC models. Model predictions generated with the coherent limit of the 3D model are compared to real DIC images acquired from imaging phantom specimens. It is shown that the model predictions resemble the real images obtained with the condenser aperture closed better than the images obtained with the aperture open. This result confirms the need for the general model that we have derived.

Publication Title

Proceedings of SPIE - The International Society for Optical Engineering

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