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Multipolarization Dark-Field Imaging of Single Endosomes in Microfluidic Neuronal Culture for Simultaneous Orientation and Displacement Tracking

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Single Molecule Microscopy in Neurobiology

Part of the book series: Neuromethods ((NM,volume 154))

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Abstract

Robust axonal transport of endosomes is critical for neuronal function. Increasingly it is appreciated that single axonal endosomes are transported by teams of molecular motors. How these motors are regulated remains poorly understood. In part, this is due to the high variability between individual endosomes. Conventional imaging approaches also only track the position of endosomes, effectively measuring one variable in a team of several motors. Here we describe how to image transport of individual axonal endosomes using multipolarization dark-field microscopy to measure both the position and the orientation of axonal endosomes. This is accomplished in a high-throughput manner by combining the imaging approach with microfluidic cell culture.

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Author information

Authors and Affiliations

  1. Department of Chemistry, Stanford University, Stanford, CA, USA

    Luke Kaplan & Bianxiao Cui

Authors
  1. Luke Kaplan
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  2. Bianxiao Cui
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Corresponding author

Correspondence to Bianxiao Cui .

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Editors and Affiliations

  1. Laboratory of Cellular and Molecular Neurobiology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan

    Nobuhiko Yamamoto

  2. Laboratory for Cell Polarity Regulation, RIKEN Center for Biosystems Dynamics Research, Suita, Osaka, Japan

    Yasushi Okada

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Kaplan, L., Cui, B. (2020). Multipolarization Dark-Field Imaging of Single Endosomes in Microfluidic Neuronal Culture for Simultaneous Orientation and Displacement Tracking. In: Yamamoto, N., Okada, Y. (eds) Single Molecule Microscopy in Neurobiology . Neuromethods, vol 154. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0532-5_8

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  • DOI: https://doi.org/10.1007/978-1-0716-0532-5_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0531-8

  • Online ISBN: 978-1-0716-0532-5

  • eBook Packages: Springer Protocols

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