Spatial mapping of protein composition and tissue organization: a primer\n for multiplexed antibody-based imaging

Abstract

Tissues and organs are composed of distinct cell types that must operate in\nconcert to perform physiological functions. Efforts to create high-dimensional\nbiomarker catalogs of these cells are largely based on transcriptomic\nsingle-cell approaches that lack the spatial context required to understand\ncritical cellular communication and correlated structural organization. To\nprobe in situ biology with sufficient coverage depth, several multiplexed\nprotein imaging methods have recently been developed. Though these\nantibody-based technologies differ in strategy and mode of immunolabeling and\ndetection tags, they commonly utilize antibodies directed against protein\nbiomarkers to provide detailed spatial and functional maps of complex tissues.\nAs these promising antibody-based multiplexing approaches become more widely\nadopted, new frameworks and considerations are critical for training future\nusers, generating molecular tools, validating antibody panels, and harmonizing\ndatasets. In this perspective, we provide essential resources and key\nconsiderations for obtaining robust and reproducible multiplexed antibody-based\nimaging data compiling specialized knowledge from domain experts and technology\ndevelopers.\n