Abstract
The recently developed ultrastructure expansion microscopy (U-ExM) technique allows us to increase the spatial resolution within a cell or tissue for microscopic imaging through the physical expansion of the sample. In this study, we validate the use of U-ExM in Trypanosoma brucei measuring the expansion factors of several different compartments/organelles and thus verify the isotropic expansion of the cell. We furthermore demonstrate the use of this sample preparation protocol for future studies by visualizing the nucleus and kDNA, as well as proteins of the cytoskeleton, the basal body, the mitochondrion and the endoplasmic reticulum. Lastly, we discuss the challenges and opportunities of U-ExM.
Highlights
Trypanosoma brucei is a unicellular flagellated protozoan parasite and the causative agent of human African sleeping sickness and Nagana in cattle
The use of ultrastructure expansion microscopy was first demonstrated in Kinetoplastea localizing a component of (a) non-expanded expanded expanded cytoskeleton kinetoplast DNA (kDNA)
We were able to increase the expansion factor to 4.2–4.6 as measured by the expansion of the kDNA, nucleus and cytoskeleton maintaining the variation at about 10%
Summary
Cite this article: Kalichava A, Ochsenreiter T. 2021 Ultrastructure expansion microscopy in Trypanosoma brucei. Cite this article: Kalichava A, Ochsenreiter T. 2021 Ultrastructure expansion microscopy in Trypanosoma brucei. Subject Area: cellular biology/molecular biology Keywords: T. brucei, expansion microscopy, centriole, kDNA, cell structure, super resolution. The recently developed ultrastructure expansion microscopy (U-ExM) technique allows us to increase the spatial resolution within a cell or tissue for microscopic imaging through the physical expansion of the sample. We validate the use of U-ExM in Trypanosoma brucei measuring the expansion factors of several different compartments/organelles and verify the isotropic expansion of the cell. We demonstrate the use of this sample preparation protocol for future studies by visualizing the nucleus and kDNA, as well as proteins of the cytoskeleton, the basal body, the mitochondrion and the endoplasmic reticulum. We discuss the challenges and opportunities of U-ExM
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