Aim

Tumor buds are tumor cells that are detaching themselves from the main tumor body and are hypothesized to be the starting point of a metastatic progression of the disease. Tumor buds are therefore a powerful biomarker in colorectal cancer. However, Currently, no phenotypic marker for tumor buds exists. Additionally, the interactions between tumor buds and their tumor microenvironment need to be investigated more thoroughly. To understand the phenotypes of tumor buds and their interacting cells, sequential Immunofluorescence (seqIF) is used to visualize up to 32 different proteins. An analysis pipeline is built to detect tumor buds and other cell types. Subsequently, unsupervised methods are used to uncover distinct subpopulations as well as enriched cell-cell interactions.