Expansion Microscopy of trichomonads

Area of Science:

• Cell Biology
• Microscopy Techniques
• Parasitology

Background:

• Recent advancements in light microscopy have focused on improving resolution in fluorescence imaging.
• Expansion Microscopy (ExM) is a technique that physically expands biological samples to enhance spatial resolution.

Purpose of the Study:

• To apply Expansion Microscopy (ExM) to the parasites Trichomonas vaginalis and Tritrichomonas foetus.
• To improve the localization and visualization of cytoskeleton-associated proteins using immunofluorescence microscopy.

Main Methods:

• Samples of Trichomonas vaginalis and Tritrichomonas foetus were embedded in a hydrophilic, swellable gel.
• The gel was expanded isotropically by 4.5 times, achieving a spatial resolution of approximately 70 nm.
• Immunofluorescence microscopy was used to visualize specific proteins, including tubulin and costain 1.

Main Results:

• ExM significantly improved the resolution of immunofluorescence imaging in both parasites.
• The structural organization of the protozoa was observed in greater detail.
• Individual microtubules of the axostyle were visualized for the first time using fluorescence microscopy.
• Detailed localization of tubulin and costain 1 was achieved, revealing new insights into the costa structure.

Conclusions:

• ExM is a valuable tool for high-resolution imaging of protozoan parasites.
• The technique enhances the study of cytoskeleton organization and protein localization in these organisms.
• ExM provides unprecedented detail for understanding parasite ultrastructure and molecular components.