Endocytosis in Trypanosoma cruzi (Euglenozoa: Kinetoplastea) epimastigotes: Visualization of ingested transferrin–gold nanoparticle complexes by confocal laser microscopy

Here we describe the visualization by confocal microscopy of ingested gold (15 nm)-labeled transferrin in epimastigote forms of the protozoan Trypanosoma cruzi. Intracellular gold labeling was evident at two sites, which represent the bottom of the cytopharynx and the reservosomes. The gold tracer was best observed by confocal microscopy by using the 633 nm excitation wavelength. Intracellular gold clusters larger than 60 nm could be visualized by either gold reflection (light scattering) or photoluminescence modes. The gold reflection mode, the gold photoluminescence mode and the anti-transferrin immunofluorescence image of gold-labeled transferrin showed co-localization, thus demonstrating that the gold visualization modes did not represent artifacts or mislocalization of the biomarker. Visualization of protein–gold nanoparticle complexes by confocal microscopy thus emerges as a promising imaging tool to explore the endocytic pathway in trypanosomes and other cell types, as well as to perform immunolocalization studies using gold-labeled secondary antibodies.