Adaptation of Iterative Ultrastructure Expansion Microscopy (iU-ExM) enables high-resolution imaging of the cytoskeleton and subcellular architecture of Trypanosoma cruzi

iU-ExM adapted for Trypanosoma cruzi epimastigotes achieves ~6.2-fold expansion in round one and ~18-fold final linear expansion, enabling confocal visualization of individual subpellicular microtubules.

• Why it matters: Kinetoplastid parasites rely on an organized microtubule network for shape, organelle positioning, and division, but conventional microscopy limits resolution of these subcellular structures.
• What they did: Cells were immunostained after an initial DHEBA hydrogel crosslinking, thermal denaturation at 85°C, neutral re-embedding, and a second BIS hydrogel, with alkaline dissolution enabling iterative expansion.
• The result: Confocal imaging after iU-ExM resolves microtubule arrangements near the basal body, flagellar pocket, Golgi complex, and cytopharynx, and links cytoskeletal reorganization to cell-cycle stages including mitotic spindle formation and cytokinesis.

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