Ultrastructure expansion microscopy in Trypanosoma brucei

Open Biol. 2021 Oct;11(10):210132. doi: 10.1098/rsob.210132. Epub 2021 Oct 13.

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.

Keywords: T. brucei; cell structure; centriole; expansion microscopy; kDNA; super resolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus / genetics
  • Cell Nucleus / ultrastructure
  • DNA, Kinetoplast / ultrastructure*
  • Microscopy, Fluorescence
  • Microtubules / metabolism
  • Mitochondria / genetics
  • Mitochondria / metabolism
  • Mitochondria / ultrastructure
  • Protozoan Proteins / ultrastructure*
  • Trypanosoma brucei brucei / genetics*
  • Trypanosoma brucei brucei / ultrastructure

Substances

  • DNA, Kinetoplast
  • Protozoan Proteins