Accurate color imaging of pathology slides using holography and absorbance spectrum estimation of histochemical stains

TL;DR

This paper introduces a holographic microscopy method that estimates absorbance spectra from multispectral images to accurately reproduce colors in pathology slides, addressing color distortion issues caused by narrow-band illumination.

Contribution

It presents a novel framework combining multispectral holography and absorbance spectrum estimation to improve color accuracy in pathology slide imaging.

Findings

Significantly reduces color errors in holographic pathology images.

Optimizes wavelength selection for accurate spectral estimation.

Applicable across various tissue and stain combinations.

Abstract

Holographic microscopy presents challenges for color reproduction due to the usage of narrow-band illumination sources, which especially impacts the imaging of stained pathology slides for clinical diagnoses. Here, an accurate color holographic microscopy framework using absorbance spectrum estimation is presented. This method uses multispectral holographic images acquired and reconstructed at a small number (e.g., three to six) of wavelengths, estimates the absorbance spectrum of the sample, and projects it onto a color tristimulus. Using this method, the wavelength selection is optimized to holographically image 25 pathology slide samples with different tissue and stain combinations to significantly reduce color errors in the final reconstructed images. The results can be used as a practical guide for various imaging applications and, in particular, to correct color distortions in holographic imaging of pathology samples spanning different dyes and tissue types.