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In Vivo Functional Imaging of the Olfactory Bulb at Single-Cell Resolution

  • Stefan Fink
  • Yury Kovalchuk
  • Ryota Homma
  • Bernd Schwendele
  • Stephan Direnberger
  • Lawrence B. Cohen
  • Oliver Griesbeck
  • Olga Garaschuk
Protocol
Part of the Neuromethods book series (NM, volume 67)

Abstract

Functional properties of neuronal circuits can be best studied in vivo in the living mammalian brain. The use of optical methods, like two-photon calcium imaging, permits analyses of network function at single-cell resolution. This chapter provides a step-by-step description of this technique. Using mouse olfactory bulb as a model system, we compare the performance of genetically encoded calcium sensor TN-XXL and small-molecule calcium indicators; describe how to choose the right calcium indicator and how to load it into the cells of interest; discuss the use of cell type-specific markers and, finally, illustrate the application of this technique for high-resolution in vivo imaging of sensory-driven neuronal activity.

Key words

Two-photon microscopy In vivo calcium imaging Multicell bolus loading Olfactory bulb Genetically encoded calcium indicators Small-molecule calcium indicators TN-XXL Fluorescence resonance energy transfer 

Notes

Acknowledgments

We thank A. Weible, S. Kasperek, G. Heck, and K. Schoentag for technical assistance. This work was supported by grants from the Deutsche Forschungsgemeinschaft (SFB 596, GA 654/1-1, SFB 870), EU FP7, and the NIH NS DC005259-39.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Stefan Fink
    • 1
  • Yury Kovalchuk
    • 1
  • Ryota Homma
    • 2
  • Bernd Schwendele
    • 1
  • Stephan Direnberger
    • 3
  • Lawrence B. Cohen
    • 2
  • Oliver Griesbeck
    • 3
  • Olga Garaschuk
    • 1
  1. 1.Department of Physiology IIUniversity of TuebingenTuebingenGermany
  2. 2.Department of PhysiologyYale UniversityNew HavenUSA
  3. 3.Max-Planck-Institute of NeurobiologyMartinsriedGermany

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