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

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...

Full description

Saved in:
Bibliographic Details
Main Authors: Clirim Jetishi, Erina A Balmer, Bianca M Berger, Carmen Faso, Torsten Ochsenreiter
Format: Article
Language:English
Published: Shared Science Publishers OG 2024-06-01
Series:Microbial Cell
Subjects:
Online Access:http://microbialcell.com/researcharticles/2024a-jetishi-microbial-cell/
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850053020676521984
author Clirim Jetishi
Erina A Balmer
Bianca M Berger
Carmen Faso
Torsten Ochsenreiter
author_facet Clirim Jetishi
Erina A Balmer
Bianca M Berger
Carmen Faso
Torsten Ochsenreiter
author_sort Clirim Jetishi
collection DOAJ
description 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.
format Article
id doaj-art-7b19c96b5d8e42dd8ecfc2bf394741de
institution DOAJ
issn 2311-2638
language English
publishDate 2024-06-01
publisher Shared Science Publishers OG
record_format Article
series Microbial Cell
spelling doaj-art-7b19c96b5d8e42dd8ecfc2bf394741de2025-08-20T02:52:38ZengShared Science Publishers OGMicrobial Cell2311-26382024-06-011119820610.15698/mic2024.06.825Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U-ExM protocolsClirim Jetishi0https://orcid.org/0000-0002-5632-7075Erina A Balmer1https://orcid.org/0000-0003-4815-2065Bianca M Berger2https://orcid.org/0000-0003-1009-4293Carmen Faso3https://orcid.org/0000-0002-1831-9365Torsten Ochsenreiter4https://orcid.org/0000-0002-8846-8526Institute of Cell Biology, University of Bern, Bern, SwitzerlandInstitute of Cell Biology, University of Bern, Bern, SwitzerlandInstitute of Cell Biology, University of Bern, Bern, SwitzerlandInstitute of Cell Biology, University of Bern, Bern, SwitzerlandInstitute of Cell Biology, University of Bern, Bern, SwitzerlandUnderstanding 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.http://microbialcell.com/researcharticles/2024a-jetishi-microbial-cell/giardia lambliaexpansion microscopysubcellular compartmentmetabolic labellingendocytosiscytoskeletonendoplasmic reticulum
spellingShingle Clirim Jetishi
Erina A Balmer
Bianca M Berger
Carmen Faso
Torsten Ochsenreiter
Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U-ExM protocols
Microbial Cell
giardia lamblia
expansion microscopy
subcellular compartment
metabolic labelling
endocytosis
cytoskeleton
endoplasmic reticulum
title Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U-ExM protocols
title_full Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U-ExM protocols
title_fullStr Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U-ExM protocols
title_full_unstemmed Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U-ExM protocols
title_short Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U-ExM protocols
title_sort expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in giardia lamblia using optimised u exm protocols
topic giardia lamblia
expansion microscopy
subcellular compartment
metabolic labelling
endocytosis
cytoskeleton
endoplasmic reticulum
url http://microbialcell.com/researcharticles/2024a-jetishi-microbial-cell/
work_keys_str_mv AT clirimjetishi expansionofmetabolicallylabelledendocyticorganellesandcytoskeletalcellstructuresingiardialambliausingoptimiseduexmprotocols
AT erinaabalmer expansionofmetabolicallylabelledendocyticorganellesandcytoskeletalcellstructuresingiardialambliausingoptimiseduexmprotocols
AT biancamberger expansionofmetabolicallylabelledendocyticorganellesandcytoskeletalcellstructuresingiardialambliausingoptimiseduexmprotocols
AT carmenfaso expansionofmetabolicallylabelledendocyticorganellesandcytoskeletalcellstructuresingiardialambliausingoptimiseduexmprotocols
AT torstenochsenreiter expansionofmetabolicallylabelledendocyticorganellesandcytoskeletalcellstructuresingiardialambliausingoptimiseduexmprotocols