Nanoresolution real-time 3D orbital tracking for studying mitochondrial trafficking in vertebrate axons in vivo
- Ludwig Maximilian University of Munich, Germany
- Technische Universität München, Germany
Abstract
We present the development and in vivo application of a feedback-based tracking microscope to follow individual mitochondria in sensory neurons of zebrafish larvae with nanometer precision and millisecond temporal resolution. By combining various technical improvements, we tracked individual mitochondria with unprecedented spatiotemporal resolution over distances of >100µm. Using these nanoscopic trajectory data, we discriminated five motional states: a fast and a slow directional motion state in both the anterograde and retrograde directions and a stationary state. The transition pattern revealed that mitochondria predominantly persist in the original direction of travel after a short pause, while transient changes of direction often exhibited longer pauses. Moreover, mitochondria in the vicinity of a second, stationary mitochondria displayed an increased probability to pause. The capability of following and optically manipulating a single organelle with high spatiotemporal resolution in a living organism offers a new approach to elucidating their function in its complete physiological context.
Data availability
The analysis software program is available on Gitlab and the wide-field images and trajectories are available on Zenodo. Source data files have been provided for all the figures.
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Wide-field Images and TrajectoriesZenodo, 10.5281/zenodo.2813946.
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Wide-field Images and TrajectoriesZenodo, 10.5281/zenodo.2815430.
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Wide-field Images and TrajectoriesZenodo, 10.5281/zenodo.281550.
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Wide-field Images and TrajectoriesZenodo, 10.5281/zenodo.2815703.
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Wide-field Images and TrajectoriesZenodo, 10.5281/zenodo.2815801.
Article and author information
Author details
Funding
Deutsche Forschungsgemeinschaft (SFB1032 (Project B3))
- Thomas Misgeld
- Don C Lamb
Fakultät für Chemie und Pharmazie, Ludwig-Maximilians-Universität München (Center for NanoScience (CeNS) and the BioImaging Network (BIN))
- Don C Lamb
H2020 European Research Council (ERC Grant Agreement n. 616791)
- Thomas Misgeld
German Center for Neurodegenerative Diseases
- Thomas Misgeld
Deutsche Forschungsgemeinschaft (research grants Mi 694/7)
- Thomas Misgeld
- Don C Lamb
Deutsche Forschungsgemeinschaft (Priority Program SPP1710)
- Thomas Misgeld
- Don C Lamb
Deutsche Forschungsgemeinschaft (SFB870 15 (Project A11))
- Thomas Misgeld
- Don C Lamb
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2019, Wehnekamp et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Nanoresolution real-time 3D orbital tracking for studying mitochondrial trafficking in vertebrate axons in vivo
eLife 8:e46059.
https://doi.org/10.7554/eLife.46059
