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Motility Analysis of Trypanosomatids.

Timothy Krüger1, Markus Engstler2

  • 1Lehrstuhl für Zell- und Entwicklungsbiologie, Theodor-Boveri-Institut, Julius-Maximilians-Universität Würzburg, Biozentrum, Am Hubland, Würzburg, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|March 30, 2020
PubMed
Summary

Analyzing microswimmer motility is now easier for diverse species using advanced microscopy. Our new protocol and tomographic approach reveal 3D movement from 2D videos.

Keywords:
Flagellar beatFlagellumIn vivo microscopyMicroswimmerMotilityProcyclicSpatiotemporal resolutionTime-dependent tomographyTrypanosome

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Area of Science:

  • Microbiology
  • Biophysics
  • Cell Biology

Background:

  • Microswimmer motility analysis is traditionally limited by microscopy resolution and cell type.
  • Recent interdisciplinary interest drives the need for advanced motility analysis techniques.
  • Diverse eukaryotic flagellates present unique challenges for in vivo motility studies.

Purpose of the Study:

  • To present a protocol for high spatiotemporal resolution motility analysis of eukaryotic flagellates.
  • To introduce a time-dependent tomographic approach for analyzing rotational locomotion.
  • To enable comprehensive 3D in vivo motility assessment from 2D videomicroscopy data.

Main Methods:

  • High spatiotemporal resolution videomicroscopy applied to diverse microswimmers.
  • Development of a novel time-dependent tomographic reconstruction method.
  • Integration of 2D microscopy data for 3D motility reconstruction.

Main Results:

  • A standardized protocol for analyzing microswimmer motility was established.
  • Rotational locomotion of periodically oscillating microswimmers was successfully demonstrated.
  • The methods allow for detailed 3D in vivo motility analysis previously unattainable.

Conclusions:

  • The described protocol and tomographic approach overcome limitations in microswimmer motility analysis.
  • This integrative method provides crucial insights into the 3D movement of diverse microswimmers.
  • The findings pave the way for more comprehensive studies of microswimmer behavior in their natural environments.