Automated pathology image classification through modern machine learning (ML) techniques in quantitative microscopy is an emerging AI application area aiming to alleviate the increased workload of pathologists and improve diagnostic accuracy and consistency. However, there are very few efforts focusing on fluorescence histology image data, which is a challenging task, not least due to the variable imaging acquisition parameters in pooled data, which can diminish the performance of ML-based decision support tools. To this end, this study introduces a harmonization preprocessing protocol for image classification within a heterogeneous fluorescence dataset in terms of image acquisition parameters and presents two state-of-the-art feature-based approaches for differentiating three classes of nuclei labelled by an expert based on (a) pathomics analysis scoring an accuracy (ACC) up to 0.957 ± 0.105, and, (b) transfer learning model exhibiting ACC up-to 0.951 ± 0.05. The proposed analysis pipelines offer good differentiation performance in the examined fluorescence histology image dataset despite the heterogeneity due to the lack of a standardized image acquisition protocol.
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