Unraveling Subcellular Ultrastructure with Cyclically Multiplexed Expansion Microscopy
Gałecki, S.; Chang, B.-J.; Zhou, F. Y.; Shen, Q.; Stoddard, D.; Chen, B.; Nicastro, D.; Fiolka, R. P.; Dean, K. M.
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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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