Electron microscopy cytochemistry and three-dimensional reconstruction of labeled structures in Trypanosoma cruzi

We report our observations by combining this approach with classical cytochemical methods to facilitate the 3D reconstruction of labeled structures and organelles

Wanderley de Souza; Carolina L. Alcantara; Narcisa L. Cunha e Silva

2020

Scholarcy highlights

  • Significant advances have occurred in the area of high-resolution scanning electron microscopy, especially related to methodologies that allow the observation of intracellular structures that are exposed either by successive abrasion with a gallium ion beam or by sectioning in epoxy-embedded cells
  • We report our observations by combining this approach with classical cytochemical methods to facilitate the 3D reconstruction of labeled structures and organelles
  • We used epimastigotes of Trypanosoma cruzi whose endocytic pathway was labeled with horseradish peroxidase, followed by fixation and detection of the peroxidase activity using the classical diaminobenzidine-osmium method followed by incubation with thiocarbohydrazide, which increases the concentration of osmium at the sites where the enzyme is located as well as the contrast of lipid-containing structures
  • All structures involved in the endocytic activity could be traced and reconstructed

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