Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U- ExM protocols

Authors:

Clirim Jetishi1,2,a, Erina A. Balmer1,2,a, Bianca M. Berger1,2,a, Carmen Faso1,3,4 and Torsten Ochsenreiter1

doi: 10.15698/mic2024.06.825
Volume 11, pp. 198 to 206, published 21/06/2024.

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Affiliations:

1 Institute of Cell Biology, University of Bern, Bern, Switzerland.

2 Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland.

3 Multidisciplinary Center for Infectious Diseases, University of Bern, Bern, Switzerland.

4 Institute of Infectious Diseases, University of Bern, Bern, Switzerland.

a Equal contribution as a first author.

Keywords: 

Giardia lamblia, expansion microscopy, subcellular compartment, metabolic labelling, endocytosis, cytoskeleton, endoplasmic reticulum.

Corresponding Author(s):

Conflict of interest statement:

All authors declare that they have no conflicts of interest.

Please cite this article as:

Clirim Jetishi, Erina A Balmer, Bianca M Berger, Carmen Faso, Torsten Ochsenreiter (2024). Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U- ExM protocols. Microbial Cell 11: 198-206. doi: 10.15698/mic2024.06.825

© 2024 Jetishi et al. This is an open-access article released under the terms of the Creative Commons Attribution (CC BY) license, which allows the unrestricted use, distribution, and reproduction in any medium, provided the original author and source are acknowledged.

Abstract:

Understanding cellular ultrastructure is tightly bound to microscopic resolution and the ability to identify individual components at that resolution. Expansion microscopy has revolutionised this topic. Here we present and compare two protocols of ultrastructure expansion microscopy that allow for 4.5-fold mostly isotropic expansion and the use of antibodies, metabolic labelling, and DNA stains to demarcate individual regions such as the endoplasmic reticulum, the nuclei, the peripheral endocytic compartments as well as the ventral disc and the cytoskeleton in Giardia lamblia. We present an optimised, shortened, and modular protocol that can be swiftly adjusted to the investigators needs in this important protozoan model organism.