<p dir="ltr">The use of spatial multiplexed imaging of tissue allows for the exploration between cellular, structural and microenvironmental relationships within a sample. Imaging systems like the Xenium Analyzer (10X Genomics) and CosMX Spatial Molecular Imager (Nanostring) are used for multiplex antibody imaging, corresponding to different proteins in multiple compartments within the tissue rather than being limited to just 3-4 wavelengths via traditional confocal imaging. Using a 19-plex panel with the Phenocycler (Akoya Biosciences), we studied spatial relationships between cells in healthy lung and fibrotic disease, using human tissue samples from control and interstitial lung disease (ILD)/idiopathic pulmonary fibrosis (IPF). The architecture of the distal lung is unique, composed of many cell types- from fine, one cell thick to cuboidal- like epithelial structures, intertwined with the corresponding endothelial layer and held open under tension to permit gaseous exchange. Using the commercial platform Visiopharm, integrated analysis protocol packages (APPs) were customised to correctly segment and identify the lung parenchyma through scripts before phenotypic exploration of healthy vs. disease.</p><p dir="ltr">Poster presented as part of the Crick BioImage Analysis Symposium 2025.</p><p dir="ltr">Permission has been given by authors to upload to Crick Figshare. Copyright remains with the original authors.</p>