Domanskyi, S.; Rubinstein, J. C.; Sheridan, T. B.; Thiesen, A.; Noorbakhsh, J.; Alcoforado Diniz, J.; Ramasamy, R.; Baker, D. S.; Sheldon, R.; Wu, Q.; Kuchel, G.; Robson, P.; Chuang, J. H.

Pathologist-guided distinctions within histology and spatial omic images provide insights into health and disease, with digital pathology leveraging artificial intelligence to automate such assessments. To train computational models, current digital pathology methods rely on upfront manual annotations, which are time-consuming to generate. Pre-annotation is poorly suited to investigating novel spatial behaviors - a major need driven by advances in spatial profiling - for which annotation criteria and data needs will be uncertain. To address these challenges, we present DIANNE, a digital pathology approach for rapid training and inference of spatial differential attributes based on train-time Positive Class Mixup Augmentation. DIANNE can compute foundation model-derived segmentation-free localization of differential classifiers across whole slide H&E images within seconds on a workstation, enabling interactive investigation of spatial niches. Predictive models can be re-trained in real-time in response to patch or regional annotation changes, clarifying determinative biological attributes across slides from only a few dozen annotated patches. We demonstrate the effectiveness of DIANNE for tumor detection, artifact identification, and exploration of pancreatic, fetal membranes and kidney tissue structures. DIANNE also provides analogous capabilities for IHC, multiplex immunofluorescence, and registered spatial transcriptomic+H&E images. DIANNE is implemented in a Jupyter toolkit, enabling rapid development of high-resolution classifiers from weakly-supervised training. DIANNE provides a practical system to quantitatively understand known and novel spatial phenotypes.