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Design and Construction of a Cost-Effective Spinning Disk System for Live Imaging of Inner Ear Tissue

  • Federico Ceriani
  • Catalin D. Ciubotaru
  • Mario BortolozziEmail author
  • Fabio Mammano
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1427)

Abstract

Confocal imaging of fluorescent probes offers a powerful, non-invasive tool which enables data collection from vast population of cells at high spatial and temporal resolution. Spinning disk confocal microscopy parallelizes the imaging process permitting the study of dynamic events in populations of living cells on the millisecond time scale. Several spinning disk microscopy solutions are commercially available, however these are often poorly configurable and relatively expensive. This chapter describes a procedure to assemble a cost-effective homemade spinning disk system for fluorescence microscopy, which is highly flexible and easily configurable. We finally illustrate a reliable protocol to obtain high-quality Ca2+ and voltage imaging data from cochlear preparations.

Key words

Confocal spinning disk microscopy Calcium imaging Voltage imaging Spontaneous activity Calcium action potentials Calcium signaling Inner hair cells Non-sensory cells Connexins 

Notes

Acknowledgments

This work was supported by Telethon Italy grants GGP13114 and GGP12269 to FM and MB, respectively.

Supplementary material

Video 1

(M4V 38728 kb)

Video 2

(MP4 16281 kb)

Video 3

(MP4 1847 kb)

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Federico Ceriani
    • 1
    • 2
    • 3
  • Catalin D. Ciubotaru
    • 4
  • Mario Bortolozzi
    • 1
    • 2
    • 3
    Email author
  • Fabio Mammano
    • 1
    • 2
    • 3
  1. 1.Department of Physics and AstronomyUniversity of PaduaPaduaItaly
  2. 2.Foundation for Advanced Biomedical ResearchVenetian Institute of Molecular Medicine (VIMM)PaduaItaly
  3. 3.CNR, Institute of Cell Biology and NeurobiologyMonterotondoItaly
  4. 4.CNR, Institute of NeurosciencePaduaItaly

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