Ultrastructure Expansion Microscopy (U-ExM) in Trypanosoma cruzi: localization of tubulin isoforms and isotypes

Parasitol Res. 2022 Oct;121(10):3019-3024. doi: 10.1007/s00436-022-07619-z. Epub 2022 Aug 9.


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.

Keywords: Acetylation; Confocal microscopy; Cytoskeleton; Mitochondria; Tyrosination.

MeSH terms

  • Animals
  • Life Cycle Stages
  • Microscopy
  • Protein Isoforms
  • Trypanosoma cruzi* / metabolism
  • Tubulin* / metabolism


  • Protein Isoforms
  • Tubulin