Key result
Ultrastructure Expansion Microscopy (U-ExM) was successfully validated in all life cycle stages of Trypanosoma cruzi, enabling the localization of tubulin isoforms and isotypes.
Why the study?
Ultrastructure Expansion Microscopy had not yet been validated or detailed in a protocol for imaging subcellular compartments and tubulin isoforms across life cycle stages of Trypanosoma cruzi.
U-ExM is a robust super-resolution method that improves the spatial resolution of T. cruzi subcellular structures without requiring highly specialized instrumentation.
Advances accessible imaging in Chagas models; leaves open validation in cardiac tissue before research adoption.
Ultrastructure Expansion Microscopy (U-ExM) is a recently developed technique that enables the increase of the spatial resolution within a cell or a tissue for microscopic imaging by physically expanding the sample. For the first time, I report a detailed protocol validating the use of U-ExM in Trypanosoma cruzi and quantifying the expansion factors of different subcellular compartments. I was able to determine the localization patterns of different tubulin isoforms, such as α-tubulin and β-tubulin. Also, I immunolocalized acetylated and tyrosinated α-tubulin isotypes in epimastigotes and use mitochondrial cell-permeable dyes to identify this organelle. Finally, U-ExM was also performed in trypomastigotes and amastigotes validating this technique in all life cycle stages of T. cruzi .
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Victoria Lucía Alonso (2022) studied Trypanosoma cruzi. Ultrastructure Expansion Microscopy (U-ExM) was evaluated on Validation of U-ExM and quantification of expansion factors of subcellular compartments. Ultrastructure Expansion Microscopy (U-ExM) was successfully validated in all life cycle stages of Trypanosoma cruzi, enabling the localization of tubulin isoforms and isotypes.
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