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Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins

Nature Methods volume 9, pages 159–172 (2012)Cite this article

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Abstract

Diverse optogenetic tools have allowed versatile control over neural activity. Many depolarizing and hyperpolarizing tools have now been developed in multiple laboratories and tested across different preparations, presenting opportunities but also making it difficult to draw direct comparisons. This challenge has been compounded by the dependence of performance on parameters such as vector, promoter, expression time, illumination, cell type and many other variables. As a result, it has become increasingly complicated for end users to select the optimal reagents for their experimental needs. For a rapidly growing field, critical figures of merit should be formalized both to establish a framework for further development and so that end users can readily understand how these standardized parameters translate into performance. Here we systematically compared microbial opsins under matched experimental conditions to extract essential principles and identify key parameters for the conduct, design and interpretation of experiments involving optogenetic techniques.

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Figure 1: Properties of depolarizing optogenetic tools.
Figure 2: Performance of depolarizing tools.
Figure 3: Properties and performance of ultrafast depolarizing tools.
Figure 4: Relationship between off kinetics and light sensitivity of optogenetic tools.
Figure 5: Properties of hyperpolarizing tools.
Figure 6: Performance of hyperpolarizing tools.

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Change history

  • 10 January 2012

    In the version of this article initially published online, the x-axis labels in Figure 5d were incorrectly labeled. The error has been corrected for the print, PDF and HTML versions of this article.

  • 10 January 2012

    In the version of this article initially published online, in the Discussion the statement "to achieve sufficient activation of cells far from the light source may require excessive hyperpolarization" was incorrect. The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

We thank A. Andalman, T. Davidson, I. Diester, S. Evans, I. Goshen, D. Mattis, I. Witten, L. Grosenick, S.-Y. Kim and C. Perry for helpful discussions, M. Lin (Stanford University) for ChIEF clones, and all memebers of the Deisseroth laboratory for their support. All viruses were packaged at University of North Carolina Vector Core. Supported by Bio-X and the Stanford Medical Scientist Training Program (J.M.), the US National Institute of Mental Health (1F32MH088010-01, K.M.T.), and the International Fulbright Science and Technology Award and a Stanford Graduate Fellowship (E.A.F.). K.D. is supported by National Institute of Mental Health, National Institute on Drug Abuse, National Institute of Neurological Disorders and Stroke, Howard Hughes Medical Institute, the Defense Advanced Research Projects Agency Reorganization and Plasticity to Accelerate Injury Recovery Program, the Keck Foundation, the McKnight Foundation and the Gatsby Charitable Foundation.

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Author notes

  1. Joanna Mattis, Kay M Tye and Emily A Ferenczi: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Bioengineering, Stanford University, Stanford, California, USA

    Joanna Mattis, Kay M Tye, Emily A Ferenczi, Charu Ramakrishnan, Daniel J O'Shea, Rohit Prakash, Lisa A Gunaydin, Minsuk Hyun, Lief E Fenno, Viviana Gradinaru, Ofer Yizhar & Karl Deisseroth

  2. Neuroscience Program, Stanford University, Stanford, California, USA

    Joanna Mattis, Emily A Ferenczi, Daniel J O'Shea, Rohit Prakash, Lisa A Gunaydin, Lief E Fenno & Karl Deisseroth

  3. CNC Program, Stanford University, Stanford, California, USA

    Viviana Gradinaru & Karl Deisseroth

  4. Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel

    Ofer Yizhar

  5. Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, California, USA

    Karl Deisseroth

  6. Howard Hughes Medical Institute, Stanford University, Stanford, California, USA

    Karl Deisseroth

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Contributions

J.M., K.M.T., E.A.F., C.R., R.P., O.Y. and K.D. contributed to study design and data interpretation. J.M. coordinated all experiments and data analysis. J.M., K.M.T., E.A.F., D.J.O., R.P. and L.E.F. contributed to acquisition of electrophysiological data. C.R. cloned all constructs, cultured primary neurons, performed transfections and managed viral packaging processes. D.J.O. wrote custom analysis scripts and analyzed all electrophysiological data. M.H. contributed to data analysis. J.M., K.M.T., C.R., L.A.G. and V.G. contributed to the histological processing and fluorescence imaging. K.D. supervised all aspects of the work. J.M., K.M.T., E.A.F. and K.D. wrote the paper.

Corresponding author

Correspondence to Karl Deisseroth.

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The authors declare no competing financial interests.

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Mattis, J., Tye, K., Ferenczi, E. et al. Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins. Nat Methods 9, 159–172 (2012). https://doi.org/10.1038/nmeth.1808

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