Imaging Giardia intestinalis cellular organisation using expansion microscopy reveals atypical centrin localisation
Year: 2024
Type: article
Abstract: Advanced imaging of microorganisms, including protists, is challenging due to their small size. Specimen expansion prior to imaging is thus beneficial to increase resolution and cellular details. Here, we present a sample preparation workflow for improved observations of the single-celled eukaryotic pathogen Giardia intestinalis (Excavata, Metamonada). The binucleated trophozoites colonize the small intestine of humans and animals and cause a diarrhoeal disease. Their remarkable morphology includes two nuclei and a pronounced microtubular cytoskeleton enabling cell motility, attachment and proliferation. By use of expansion and confocal microscopy, we resolved in a great detail subcellular structures and organelles of the parasite cell. The acquired spatial resolution enabled novel observations of centrin localization at Giardia basal bodies. Interestingly, non-luminal centrin localization between the Giardia basal bodies was observed, which is an atypical eukaryotic arrangement. Our protocol includes antibody staining and can be used for the localization of epitope-tagged proteins, as well as for differential organelle labelling by amino reactive esters. This fast and simple technique is suitable for routine use without a superresolution microscopy equipment. • Expansion microscopy (ExM) is suitable for imaging parasitic protists, such as Giardia intestinalis (Metamonada), to overcome limitations imposed by tiny size. • The ExM pipeline presented here enables high-resolution imaging of the structural organisation of Giardia cells without a need of a superresolution device. • Novel observations regarding a more precise centrin localization in Giardia trophozoites are presented. • Centrin is in Giardia localized among and around basal bodies of its eight flagella, and is absent from the basal body lumen.
Source: Experimental Parasitology
Language: English
Subfield: Parasitology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $3,340
OpenAlex ID: W4402315541