Spleen Tissue Segmentation Algorithm for Cryo-Imaging Data

Spleen tissue segmentation is an essential process for analyzing various immunological diseases as observed in the cryo-imaging data. Because manual labeling of the spleen tissue by human experts is not efficient, an automatic segmentation algorithm is needed. In this study, we developed a novel algorithm for automatically segmenting spleen substructures including white pulp and red pulp for the first time. The algorithm is designed for datasets created by a cryo-imaging system. This unique technology can effectively enable cellular tracking anywhere in the whole mouse with single-cell sensitivity. The proposed algorithm consists of four components: initial spleen mask creation, feature extraction, Supervised Patch-based Fuzzy c-Mean (spFCM) classification, and post-processing. The algorithm accurately and efficiently labeled spleen tissues in all experiment settings. The algorithm also improved the spleen segmentation throughput by 90 folds as compared to the manual segmentation. Moreover, we show that our novel spFCM algorithm outperformed traditional fast-learning classifiers as well as the U-Net deep-learning model in many aspects. Two major contributions of this paper are (1) an explainable algorithm for segmenting spleen tissues in cryo-images for the first time and (2) an spFCM algorithm as a new classifier. We also discussed that our work can be beneficial to researchers who work not only in the fields of graft-versus-host disease (GVHD) mouse models, but also in that of other immunological disease models where spleen analysis is essential. Future work building upon our research may lay the foundations for biomedical studies that utilize cryo-imaging technology.