Unraveling Subcellular Ultrastructure with Cyclically Multiplexed Expansion Microscopy
Seweryn GałeckiiD, Bo-Jui ChangiD, Felix ZhouiD, Qionghua SheniD, Daniel StoddardiD, Bingying CheniD, Daniela NicastroiD, Reto Paul FiolkaiD, Kevin M. DeaniD
Abstract
Despite advances in fluorescence microscopy, spectral overlap and limited resolution hinder the dense mapping of the cellular ultrastructure. To overcome these challenges, we developed Cy-ExM, a high-plex imaging strategy that integrates optimized cryo-fixation for antigen preservation, expansion microscopy, and iterative immunofluorescence labeling. Using oblique plane microscopy, we perform three-dimensional super-resolution imaging of 20 biological targets encompassing the full cellular volume of individual mammalian cells.
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