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Patterned Two-Photon Illumination for High-Speed Functional Imaging of Brain Networks In Vivo

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Advanced Optical Methods for Brain Imaging

Part of the book series: Progress in Optical Science and Photonics ((POSP,volume 5))

Abstract

Two-photon functional imaging combined with optogenetic manipulation represents a powerful tool to investigate neural network function and how the activity of specific circuits drives behavior. To fulfill these goals, imaging methods should ideally have a temporal resolution close to the temporal scale of neural activity (i.e., in the millisecond range). Classical laser-scanning two-photon microscopy, however, relies on beam deflection by galvanometric mirrors whose inertia, as well as the time the beam dwells on each image pixel, severely limits its temporal resolution. In this chapter, we focus on one approach to high-speed fluorescence recording, namely scanless imaging using patterned two-photon illumination. We will discuss recent evidence showing that this technique can be used to perform high-speed (up to 1 kHz) recording of calcium signals hundreds of micrometers deep in the rodent brain. Moreover, we describe the combination of scanless imaging with single-photon optogenetic manipulation for all-optical readout and perturbation of brain networks and with microendoscopic imaging for high-speed monitoring of circuit dynamics in deeper brain areas. Finally, we discuss the advantages and limitations of scanless imaging as well as possible future technical improvements that will likely contribute to significantly increasing our understanding of the role of neural networks in driving brain function and behavior.

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Acknowledgements

We thank members of the Fellin laboratory for helpful comments on the manuscript. This work was supported by the ERC (NEURO-PATTERNS), NIH (1U01NS090576-01), FP7 (DESIRE), and, in part, from Flag-Era JTC Human Brain Project (SLOW-DYN) to TF.

Competing financial interests The authors declare no competing financial interests.

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Authors and Affiliations

  1. Optical Approaches to Brain Function Laboratory, Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genoa, Italy

    Serena Bovetti, Claudio Moretti & Tommaso Fellin

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  1. Serena Bovetti
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  2. Claudio Moretti
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  3. Tommaso Fellin
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Correspondence to Tommaso Fellin .

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  1. Institute of Biophotonics, National Yang-Ming University, Taipei, Taiwan

    Fu-Jen Kao

  2. Department of Electrical and Computer Engineering, Boston University, Boston, Massachusetts, USA

    Gerd Keiser

  3. Institute of Biophotonics, National Yang-Ming University, Taipei, Taiwan

    Ankur Gogoi

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Bovetti, S., Moretti, C., Fellin, T. (2019). Patterned Two-Photon Illumination for High-Speed Functional Imaging of Brain Networks In Vivo. In: Kao, FJ., Keiser, G., Gogoi, A. (eds) Advanced Optical Methods for Brain Imaging. Progress in Optical Science and Photonics, vol 5. Springer, Singapore. https://doi.org/10.1007/978-981-10-9020-2_7

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  • DOI: https://doi.org/10.1007/978-981-10-9020-2_7

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