Nanoresolution real-time 3D orbital tracking for studying mitochondrial trafficking in vertebrate axons in vivo

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 mitocho...

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Bibliographic Details
Published in:eLife 2019-06, Vol.8
Main Authors: Wehnekamp, Fabian, Plucińska, Gabriela, Thong, Rachel, Misgeld, Thomas, Lamb, Don C
Format: Article
Language:English
Subjects:
Algorithms
Animals
Anterograde transport
Axons
Axons - metabolism
Biological Transport
Cell Tracking - methods
Coordinate transformations
Electronics
fluorescence
Homeostasis
Imaging, Three-Dimensional - methods
Larva - metabolism
Microscopy, Confocal - methods
Mitochondria
Mitochondria - metabolism
mitochondria trafficking
Motility
Nanotechnology - methods
Neural circuitry
Neurons
Physics of Living Systems
Physiological aspects
Physiology
Retrograde transport
Sensory neurons
Sensory Receptor Cells - metabolism
single particle tracking
Software
Structural Biology and Molecular Biophysics
Tools and Resources
Vertebrates - metabolism
Zebrafish
Zebrafish - metabolism
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Summary: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 >100 µm. 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, after a pause, mitochondria predominantly persist in the original direction of travel, 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.
ISSN:2050-084X
2050-084X
DOI:10.7554/elife.46059