Aptamer-based expansion microscopy platform enables signal-amplified imaging of dendritic spines

Super-resolution imaging of dendritic spines (DS) can provide valuable information for mechanistic studies related to synaptic physiology and neural plasticity, but challenged by their small dimension (50–200 nm) below the spatial resolution of conventional optical microscopes. In this work, by combining the molecular recognition specificity of aptamer with high programmability of DNA nanotechnology, we developed an expansion microscopy (ExM) platform for imaging DS with enhanced spatial resolution and amplified signal output. Our results demonstrated that the aptamer probe could specifically bind to DS of primary hippocampal neurons. With physical expansion, the DS structure could be effectively enlarged by 4–5 folds, leading to the generation of more structural information. Meantime, the aptamer binding signal could be readily amplified by the introduction of DNA signal amplification strategy, overcoming the drawback of fluorescence dilution during the ExM treatment. This platform enabled evaluation of ischemia-induced early stroke based on the morphological change of DS, highlighting a promising avenue for studying nanoscale structures in biological systems. [Display omitted] •We developed an aptamer-HCR-based ExM platform for super-resolution imaging of dendritic spines with enhanced sensitivity.•The signal output could be enhanced to address the drawback of signal dilution during the physical expansion process.•Our platform allowed super-resolution of morphological changes of dendritic spines in the early stage of stroke.