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Ultrastructure expansion microscopy in Trypanosoma brucei.

Open biology (2021-10-13)
Ana Kalichava, Torsten Ochsenreiter
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.

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Sigma-Aldrich
Paraformaldehyde, reagent grade, crystalline
Sigma-Aldrich
Anti-Guinea Pig IgG (H+L), CF 647 antibody produced in goat, ~2 mg/mL, affinity isolated antibody