Spatial resolution of Alzheimer’s disease using automated high-plex immunofluorescence 3556

Abstract Description Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by amyloid-beta plaques, neurofibrillary tangles, neuronal loss, and cognitive impairment. While our understanding of AD has evolved, mapping the cellular and molecular interplay between neurodegeneration and neuroimmunology remains challenging. Recent innovations in spatial biology have enabled multiple advances through the detailed identification of cell lineages and intercellular interactions within the native architecture of many tissue types. In this study, we leveraged a spatial proteomics approach to map the cellular interactions underlying neurodegeneration in AD. Formalin-fixed paraffin-embedded tissue sections of human AD and healthy brain samples were stained and imaged using the fully-automated sequential immunofluorescence (seqIF™) workflow on COMET™. Images were analyzed by HORIZON™ software to reveal in situ single-cell level features. We designed a highplex proteomics panel to characterize the spatial localization of AD-associated protein aggregates, along with key cell populations such as neurons, astrocytes, microglia, and vasculature-associated cells in the same tissue section. Our analysis identified inflammatory activation of microglia and astrocytes in the proximity of amyloid-beta plaques. This spatial approach offers novel insights into the spatial relationships between neurodegenerative lesions and the neighboring cells, shedding light on the neuroimmune landscape of AD. Topic Categories Neuroimmunology (NEUR)