Skip to main content
SpringerLink
Log in

Three-Color Simultaneous Live Imaging of Autophagy-Related Structures

  • Protocol
  • First Online:
Autophagy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1880))

Abstract

Simultaneous live cell imaging of multiple proteins helps to analyze mobility and interactions among proteins over time. Since autophagosomes depend on other organelles for their formation, it is necessary to observe this process with multiple fluorsphores to mark multiple organelles and the autophagosomes. To do so, we set up three cameras on one microscope to be able to acquire three colors at the same time. Here we describe the setup using a Yokogawa spinning disk confocal microscope (CSU-W1) with Andor TuCam system attaching 3 × Zyla 4.2 CMOS cameras (Andor) and detail the method for acquiring live images.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 189.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675

    Article  CAS  Google Scholar 

  2. Miura K, Rueden C, Hiner M, Schindelin J, Riefdort J (2014) ImageJ Plugin CorrectBleach V2.0.2. Zenodo. https://doi.org/10.5281/zenodo.30769

Download references

Author information

Authors and Affiliations

  1. Department of Genetics, Osaka University Graduate School of Medicine, Osaka, Japan

    Hiroyuki Ueda & Ouin Kunitaki

  2. Department of Intracellular Membrane Dynamics, Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan

    Hiroyuki Ueda

  3. Andor Technology Ltd., Newark, DE, USA

    Maho Hamasaki

Authors
  1. Hiroyuki Ueda
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ouin Kunitaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maho Hamasaki
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Signalling Programme, Babraham Institute, Cambridge, UK

    Nicholas Ktistakis

  2. Signalling Programme, Babraham Institute, Cambridge, UK

    Oliver Florey

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Ueda, H., Kunitaki, O., Hamasaki, M. (2019). Three-Color Simultaneous Live Imaging of Autophagy-Related Structures. In: Ktistakis, N., Florey, O. (eds) Autophagy. Methods in Molecular Biology, vol 1880. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8873-0_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8873-0_14

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8872-3

  • Online ISBN: 978-1-4939-8873-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation