Imaging Giardia intestinalis cellular organisation using expansion microscopy reveals atypical centrin localisation
doi: 10.1016/j.exppara.2024.108831.
Online ahead of print.
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Exp Parasitol.
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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 localisation 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 localisation 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.
Keywords:
Giardia; caltractin; centrin; cytoskeleton; expansion microscopy; protist.
Copyright © 2024. Published by Elsevier Inc.
Conflict of interest statement
Declaration of Competing Interest The authors of Imaging Giardia intestinalis cellular organisation using expansion microscopy reveals atypical centrin localisation declare no conflict of interests.
Fonte original PubMed