SPECTREPlex: an automated, fast, high-resolution enabled approach for multiplexed cyclic imaging and tissue spatial analysis.

Mapping the spatial organization of tissues is critical to understanding organ biology in health and disease. Developments in multiplexed antibody-based, fluorescence labelling methods have provided unique insights into tissue microenvironments. However, many current methods have a variety of limitations that reduce their practical utilization including cost, time and technical complexity. To address these drawbacks, we developed Spatial Photo-inactivation Enhanced Cyclic Target REsolved multiPlexing (SPECTRE-Plex). SPECTRE-Plex is a relatively low cost, end-to-end technique based on a series of methods that significantly improve speed, automation, and resolution of cyclic multiplex immunofluorescence imaging. We describe a representative example of the application of the method by investigating spatial cellular and neighborhood changes in the proximal small intestine between healthy tissue and active celiac disease.