Abstract

We present an investigation of the impact of partial coherence on optical imaging systems with the focus on whole slide imaging (WSI) systems for digital pathology. The investigation is based on the analysis of the edge response of the optical system, which gives rise to an apparent optical transfer function (OTF) that can be linked to two elementary complex functions Q and U. The function Q is directly related to the transmission cross coefficient (TCC) and can be identified with the performance function first introduced by Kintner and Sillitto. The function U depends on the TCC in a more involved way. When there are no aberrations the Q-function corresponds to the real part of the apparent OTF and the U function to the imaginary part of the apparent OTF. Close to the incoherent limit the effect of the U function is a mere shift of the edge compared to the fully incoherent case. We propose a new expression for the dependence of the depth of focus (DOF) on spatial frequency and on the partial coherence factor σ, and validate it by simulation. Partial coherence effects are investigated experimentally on a WSI system with a compact LED-based Köhler illumination unit with variable condenser NA. This unit incorporates a top hat diffuser for providing a reasonably uniform illumination field, with variations below 10% across the imaged field of view. The measurements of the apparent through-focus OTF derived from edges on a custom resolution chart for different σ were substantially in agreement with the simulations. Finding an optimal value for σ is not straightforward as lateral resolution and the level of edge ringing improve with increasing σ, whereas edge contrast and DOF improve with decreasing σ. We assess that the trade-off for the particular application of WSI systems for digital pathology is optimized for a σ value in the range of 0.55-0.75.

Highlights

  • Partial coherence influences image formation in microscopy via the partial coherence factor σ = NAill/NA, the ratio between the illumination Numerical Aperture NAill and the imaging NA [1, 2, 3]

  • We present an investigation of the impact of partial coherence on optical imaging systems with the focus on Whole Slide Imaging (WSI) systems for digital pathology

  • A typical example of the measured edge response is shown in Fig.13, which shows measured intensity profiles at the center of the Field Of View (FOV) of the objective lens for tangentially oriented edges

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Summary

Introduction

Partial coherence influences image formation in microscopy via the partial coherence factor σ = NAill/NA, the ratio between the illumination (condenser) Numerical Aperture NAill and the imaging (objective) NA [1, 2, 3]. The sharpness in the image, that can be quantified by the steepness of the edge response, improves towards the coherent limit. This beneficial effect, is accompanied by edge ringing artefacts. A general rule of thumb in the field of microscopy is to slightly sacrifice resolution for image sharpness by reducing the illumination NAill to about 0.75 times the imaging NA

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