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Autophagy pp 223-230 | Cite as

Three-Color Simultaneous Live Imaging of Autophagy-Related Structures

  • Hiroyuki Ueda
  • Ouin Kunitaki
  • Maho Hamasaki
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Live cell imaging Three cameras 

References

  1. 1.
    Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675CrossRefGoogle Scholar
  2. 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

Copyright information

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

Authors and Affiliations

  • Hiroyuki Ueda
    • 1
    • 2
  • Ouin Kunitaki
    • 1
  • Maho Hamasaki
    • 3
  1. 1.Department of GeneticsOsaka University Graduate School of MedicineOsakaJapan
  2. 2.Department of Intracellular Membrane DynamicsGraduate School of Frontier Biosciences, Osaka UniversityOsakaJapan
  3. 3.Andor Technology Ltd.NewarkUSA

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