Automating tumor-infiltrating lymphocyte assessment in breast cancer histopathology images using QuPath: a transparent and accessible machine learning pipeline

In this study, we built an end-to-end tumor-infiltrating lymphocytes (TILs) assessment pipeline within QuPath, demonstrating the potential of easily accessible tools to perform complex tasks in a fully automatic fashion. First, we trained a pixel classifier to segment tumor, tumor-associated stroma, and other tissue compartments in breast cancer H&E-stained whole-slide images (WSI) to isolate tumor-associated stroma for subsequent analysis. Next, we applied a pre-trained StarDist deep learning model in QuPath for cell detection and used the extracted cell features to train a binary classifier distinguishing TILs from other cells. To evaluate our TILs assessment pipeline, we calculated the TIL density in each WSI and categorized them as low, medium, or high TIL levels. Our pipeline was evaluated against pathologist-assigned TIL scores, achieving a Cohen's kappa of 0.71 on the external test set, corroborating previous research findings. These results confirm that existing software can offer a practical solution for the assessment of TILs in H&E-stained WSIs of breast cancer.

Open-source framework for detecting bias and overfitting for large pathology images

Even foundational models that are trained on datasets with billions of data samples may develop shortcuts that lead to overfitting and bias. Shortcuts are non-relevant patterns in data, such as the background color or color intensity. So, to ensure the robustness of deep learning applications, there is a need for methods to detect and remove such shortcuts. Today's model debugging methods are time consuming since they often require customization to fit for a given model architecture in a specific domain. We propose a generalized, model-agnostic framework to debug deep learning models. We focus on the domain of histopathology, which has very large images that require large models - and therefore large computation resources. It can be run on a workstation with a commodity GPU. We demonstrate that our framework can replicate non-image shortcuts that have been found in previous work for self-supervised learning models, and we also identify possible shortcuts in a foundation model. Our easy to use tests contribute to the development of more reliable, accurate, and generalizable models for WSI analysis. Our framework is available as an open-source tool available on github.

A Lightweight and Extensible Cell Segmentation and Classification Model for Whole Slide Images

Developing clinically useful cell-level analysis tools in digital pathology remains challenging due to limitations in dataset granularity, inconsistent annotations, high computational demands, and difficulties integrating new technologies into workflows. To address these issues, we propose a solution that enhances data quality, model performance, and usability by creating a lightweight, extensible cell segmentation and classification model. First, we update data labels through cross-relabeling to refine annotations of PanNuke and MoNuSAC, producing a unified dataset with seven distinct cell types. Second, we leverage the H-Optimus foundation model as a fixed encoder to improve feature representation for simultaneous segmentation and classification tasks. Third, to address foundation models' computational demands, we distill knowledge to reduce model size and complexity while maintaining comparable performance. Finally, we integrate the distilled model into QuPath, a widely used open-source digital pathology platform. Results demonstrate improved segmentation and classification performance using the H-Optimus-based model compared to a CNN-based model. Specifically, average $R^2$ improved from 0.575 to 0.871, and average $PQ$ score improved from 0.450 to 0.492, indicating better alignment with actual cell counts and enhanced segmentation quality. The distilled model maintains comparable performance while reducing parameter count by a factor of 48. By reducing computational complexity and integrating into workflows, this approach may significantly impact diagnostics, reduce pathologist workload, and improve outcomes. Although the method shows promise, extensive validation is necessary prior to clinical deployment.

Fast TILs estimation in lung cancer WSIs based on semi-stochastic patch sampling

Addressing the critical need for accurate prognostic biomarkers in cancer treatment, quantifying tumor-infiltrating lymphocytes (TILs) in non-small cell lung cancer (NSCLC) presents considerable challenges. Manual TIL quantification in whole slide images (WSIs) is laborious and subject to variability, potentially undermining patient outcomes. Our study introduces an automated pipeline that utilizes semi-stochastic patch sampling, patch classification to retain prognostically relevant patches, and cell quantification using the HoVer-Net model to streamline the TIL evaluation process. This pipeline efficiently excludes approximately 70% of areas not relevant for prognosis and requires only 5% of the remaining patches to maintain prognostic accuracy (c-index 0.65 +- 0.01). The computational efficiency achieved does not sacrifice prognostic accuracy, as demonstrated by the TILs score's strong correlation with patient survival, which surpasses traditional CD8 IHC scoring methods. While the pipeline demonstrates potential for enhancing NSCLC prognostication and personalization of treatment, comprehensive clinical validation is still required. Future research should focus on verifying its broader clinical utility and investigating additional biomarkers to improve NSCLC prognosis.