Article | Published:

Reconstruction of firing rate changes across neuronal populations by temporally deconvolved Ca2+ imaging

Nature Methods volume 3, pages 377383 (2006) | Download Citation

Subjects

Abstract

Methods to record action potential (AP) firing in many individual neurons are essential to unravel the function of complex neuronal circuits in the brain. A promising approach is bolus loading of Ca2+ indicators combined with multiphoton microscopy. Currently, however, this technique lacks cell-type specificity, has low temporal resolution and cannot resolve complex temporal firing patterns. Here we present simple solutions to these problems. We identified neuron types by colocalizing Ca2+ signals of a red-fluorescing indicator with genetically encoded markers. We reconstructed firing rate changes from Ca2+ signals by temporal deconvolution. This technique is efficient, dramatically enhances temporal resolution, facilitates data interpretation and permits analysis of odor-response patterns across thousands of neurons in the zebrafish olfactory bulb. Hence, temporally deconvolved Ca2+ imaging (TDCa imaging) resolves limitations of current optical recording techniques and is likely to be widely applicable because of its simplicity, robustness and generic principle.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , , & Optical imaging of neuronal activity. Physiol. Rev. 68, 1285–1366 (1988).

  2. 2.

    et al. Imaging membrane potential with voltage-sensitive dyes. Biol. Bull. 198, 1–21 (2000).

  3. 3.

    & On the nature of the BOLD fMRI contrast mechanism. Magn. Reson. Imaging 22, 1517–1531 (2004).

  4. 4.

    , & Two-photon laser scanning fluorescence microscopy. Science 248, 73–76 (1990).

  5. 5.

    & Deep tissue two-photon microscopy. Nat Methods 2, 932–940 (2005).

  6. 6.

    , , & In vivo two-photon calcium imaging of neuronal networks. Proc. Natl. Acad. Sci. USA 100, 7319–7324 (2003).

  7. 7.

    , , , & “In vivo” monitoring of neuronal network activity in zebrafish by two-photon Ca2+ imaging. Pflugers Arch. 446, 766–773 (2003).

  8. 8.

    , , , & Functional imaging with cellular resolution reveals precise micro-architecture in visual cortex. Nature 433, 597–603 (2005).

  9. 9.

    et al. Early development of functional spatial maps in the zebrafish olfactory bulb. J. Neurosci. 25, 5784–5795 (2005).

  10. 10.

    , , , & In vivo calcium imaging of circuit activity in cerebellar cortex. J. Neurophysiol. 94, 1636–1644 (2005).

  11. 11.

    , & Imaging input and output of neocortical networks in vivo. Proc. Natl Acad. Sci. USA 102, 14063–14068 (2005).

  12. 12.

    , & Ca2+ buffering and action potential-evoked Ca2+ signaling in dendrites of pyramidal neurons. Biophys. J. 70, 1069–1081 (1996).

  13. 13.

    Olfactory network dynamics and the coding of multidimensional signals. Nat. Rev. Neurosci. 3, 884–895 (2002).

  14. 14.

    & Functional organization of olfactory system. J. Neurobiol. 30, 123–176 (1996).

  15. 15.

    et al. A highly conserved enhancer in the Dlx5/Dlx6 intergenic region is the site of cross-regulatory interactions between Dlx genes in the embryonic forebrain. J. Neurosci. 20, 709–721 (2000).

  16. 16.

    , , & Imaging neuronal activity during zebrafish behavior with a genetically encoded calcium indicator. J. Neurophysiol. 90, 3986–3997 (2003).

  17. 17.

    & Dynamic optimization of odor representations in the olfactory bulb by slow temporal patterning of mitral cell activity. Science 291, 889–894 (2001).

  18. 18.

    A novel single-cell staining procedure performed in vivo under electrophysiological control: morpho-functional features of juxtacellularly labeled thalamic cells and other central neurons with biocytin or Neurobiotin. J. Neurosci. Methods 65, 113–136 (1996).

  19. 19.

    & Combinatorial and chemotopic odorant coding in the zebrafish olfactory bulb visualized by optical imaging. Neuron 18, 737–752 (1997).

  20. 20.

    & Dynamics of olfactory bulb input and output activity during odor stimulation in zebrafish. J. Neurophysiol. 91, 2658–2669 (2004).

  21. 21.

    , & Multiplexing using synchrony in the zebrafish olfactory bulb. Nat. Neurosci. 7, 862–871 (2004).

  22. 22.

    & Optical imaging and control of genetically designated neurons in functioning circuits. Annu. Rev. Neurosci. 28, 533–563 (2005).

  23. 23.

    , , & Internal dynamics determine the cortical response to thalamic stimulation. Neuron 48, 811–823 (2005).

  24. 24.

    & Odor- and context-dependent modulation of mitral cell activity in behaving rats. Nat. Neurosci. 2, 1003–1009 (1999).

  25. 25.

    & Matching patterns of activity in primate prefrontal area 8a and parietal area 7ip neurons during a spatial working memory task. J. Neurophysiol. 79, 2919–2940 (1998).

  26. 26.

    , & Natural waking and sleep states: a view from inside neocortical neurons. J. Neurophysiol. 85, 1969–1985 (2001).

  27. 27.

    , & Transformation of olfactory representations in the Drosophila antennal lobe. Science 303, 366–370 (2004).

  28. 28.

    Neuronal synchrony: a versatile code for the definition of relations? Neuron 24, 49–65 (1999).

  29. 29.

    , , , & Two-photon calcium imaging reveals an odor-evoked map of activity in the fly brain. Cell 112, 271–282 (2003).

  30. 30.

    & The coding of odour-intensity in the honeybee antennal lobe: local computation optimizes odour representation. Eur. J. Neurosci. 18, 2119–2132 (2003).

Download references

Acknowledgements

We thank W. Denk, T. Euler, J. Kerr, G. Laurent, H. Riecke, P.H. Seeburg and members of the Friedrich laboratory for support, helpful discussions, and/or comments on the manuscript. This work was supported by the Max Planck-Society, the Deutsche Forschungsgemeinschaft (DFG; SFB 488), and a fellowship from the Boehringer Ingelheim Fonds to E.Y.

Author information

Affiliations

  1. Max Planck Institute for Medical Research, Department of Biomedical Optics, Jahnstr. 29, 69120 Heidelberg, Germany.

    • Emre Yaksi
    •  & Rainer W Friedrich

Authors

  1. Search for Emre Yaksi in:

  2. Search for Rainer W Friedrich in:

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Rainer W Friedrich.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Principle of firing rate reconstruction by temporal deconvolution.

  2. 2.

    Supplementary Fig. 2

    Decay time constants and their influence on deconvolution.

  3. 3.

    Supplementary Fig. 3

    Iterative smooting procedure.

  4. 4.

    Supplementary Fig. 4

    Deconvolution parameter search results.

  5. 5.

    Supplementary Fig. 5

    Comparison of temporal deconvolution to other methods.

  6. 6.

    Supplementary Methods

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nmeth874

Further reading