Staining normalization in histopathology: Method benchmarking using multicenter dataset

Hematoxylin and Eosin (H&E) has been the gold standard in tissue analysis for decades, however, tissue specimens stained in different laboratories vary, often significantly, in appearance. This variation poses a challenge for both pathologists' and AI-based downstream analysis. Minimizing stain variation computationally is an active area of research. To further investigate this problem, we collected a unique multi-center tissue image dataset, wherein tissue samples from colon, kidney, and skin tissue blocks were distributed to 66 different labs for routine H&E staining. To isolate staining variation, other factors affecting the tissue appearance were kept constant. Further, we used this tissue image dataset to compare the performance of eight different stain normalization methods, including four traditional methods, namely, histogram matching, Macenko, Vahadane, and Reinhard normalization, and two deep learning-based methods namely CycleGAN and Pixp2pix, both with two variants each. We used both quantitative and qualitative evaluation to assess the performance of these methods. The dataset's inter-laboratory staining variation could also guide strategies to improve model generalizability through varied training data

Improving Performance in Colorectal Cancer Histology Decomposition using Deep and Ensemble Machine Learning

In routine colorectal cancer management, histologic samples stained with hematoxylin and eosin are commonly used. Nonetheless, their potential for defining objective biomarkers for patient stratification and treatment selection is still being explored. The current gold standard relies on expensive and time-consuming genetic tests. However, recent research highlights the potential of convolutional neural networks (CNNs) in facilitating the extraction of clinically relevant biomarkers from these readily available images. These CNN-based biomarkers can predict patient outcomes comparably to golden standards, with the added advantages of speed, automation, and minimal cost. The predictive potential of CNN-based biomarkers fundamentally relies on the ability of convolutional neural networks (CNNs) to classify diverse tissue types from whole slide microscope images accurately. Consequently, enhancing the accuracy of tissue class decomposition is critical to amplifying the prognostic potential of imaging-based biomarkers. This study introduces a hybrid Deep and ensemble machine learning model that surpassed all preceding solutions for this classification task. Our model achieved 96.74% accuracy on the external test set and 99.89% on the internal test set. Recognizing the potential of these models in advancing the task, we have made them publicly available for further research and development.

Domain-specific transfer learning in the automated scoring of tumor-stroma ratio from histopathological images of colorectal cancer

Tumor-stroma ratio (TSR) is a prognostic factor for many types of solid tumors. In this study, we propose a method for automated estimation of TSR from histopathological images of colorectal cancer. The method is based on convolutional neural networks which were trained to classify colorectal cancer tissue in hematoxylin-eosin stained samples into three classes: stroma, tumor and other. The models were trained using a data set that consists of 1343 whole slide images. Three different training setups were applied with a transfer learning approach using domain-specific data i.e. an external colorectal cancer histopathological data set. The three most accurate models were chosen as a classifier, TSR values were predicted and the results were compared to a visual TSR estimation made by a pathologist. The results suggest that classification accuracy does not improve when domain-specific data are used in the pre-training of the convolutional neural network models in the task at hand. Classification accuracy for stroma, tumor and other reached 96.1$\%$ on an independent test set. Among the three classes the best model gained the highest accuracy (99.3$\%$) for class tumor. When TSR was predicted with the best model, the correlation between the predicted values and values estimated by an experienced pathologist was 0.57. Further research is needed to study associations between computationally predicted TSR values and other clinicopathological factors of colorectal cancer and the overall survival of the patients.