Graphical and interactive spatial proteomics image analysis workflow

Spatial proteomics provides a spatially resolved view of protein expression and localization within cells and tissues by mapping the location and abundance of proteins. There is a need for fully-integrated end-to-end imaging workflows for spatial proteomic analysis that are flexible, reproducible, and support graphical and interactive visualizations. We present a modular and interactive spatial proteomic image analysis workflow with individual containerized steps that empowers biomedical researchers to reproducibly execute and customize complex analyses. Our workflow consists of cell segmentation, unsupervised clustering with optional batch correction, validation of clusters on the image, and cell type clustering results visualization. A form-based graphical interface can be utilized to execute and customize multi-step workflows with a single click or interactively adjust image processing steps within the workflow, apply workflows to various datasets, and modify input parameters as needed. We illustrated the functionality of our workflow using human normal tonsil and colorectal cancer tissues stained by high-plex immunohistochemistry.