UNIT 2.14 High-Speed Multineuron Calcium Imaging Using Nipkow-Type Confocal Microscopy

  1. Naoya Takahashi1,
  2. Shigeyuki Oba2,
  3. Naoto Yukinawa2,
  4. Sakiko Ujita1,
  5. Mika Mizunuma1,
  6. Norio Matsuki1,
  7. Shin Ishii2,
  8. Yuji Ikegaya1

Published Online: 1 OCT 2011

DOI: 10.1002/0471142301.ns0214s57

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Takahashi, N., Oba, S., Yukinawa, N., Ujita, S., Mizunuma, M., Matsuki, N., Ishii, S. and Ikegaya, Y. 2011. High-Speed Multineuron Calcium Imaging Using Nipkow-Type Confocal Microscopy. Current Protocols in Neuroscience. 57:2.14:2.14.1–2.14.10.

Author Information

  1. 1

    Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan

  2. 2

    Laboratory for Integrated Systems Biology, Graduate School of Informatics, Kyoto University, Kyoto, Japan

Publication History

  1. Published Online: 1 OCT 2011
  2. Published Print: OCT 2011


Conventional confocal and two-photon microscopy scan the field of view sequentially with single-point laser illumination. This raster-scanning method constrains video speeds to tens of frames per second, which are too slow to capture the temporal patterns of fast electrical events initiated by neurons. Nipkow-type spinning-disk confocal microscopy resolves this problem by the use of multiple laser beams. We describe experimental procedures for functional multineuron calcium imaging (fMCI) based on Nipkow-disk confocal microscopy, which enables us to monitor the activities of hundreds of neurons en masse at a cellular resolution at up to 2000 fps. Curr. Protoc. Neurosci. 57:2.14.1-2.14.10. © 2011 by John Wiley & Sons, Inc.


  • imaging;
  • microscopy;
  • calcium;
  • neuron;
  • spike