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  Two-photon imaging of neuronal populations in the visual cortex of freely-moving animals

Sawinski, J., Wallace, D., Greenberg, D., Grossmann, S., Denk, W., & Kerr, J. (2009). Two-photon imaging of neuronal populations in the visual cortex of freely-moving animals. Poster presented at 39th Annual Meeting of the Society for Neuroscience (Neuroscience 2009), Chicago, IL, USA.

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Sawinski, J1, 2, 3, Author              
Wallace, DJ1, 2, 3, Author              
Greenberg, DS1, 2, 3, Author              
Grossmann, S, Author
Denk, W, Author
Kerr, JN1, 2, 3, Author              
Affiliations:
1Former Research Group Network Imaging, Max Planck Institute for Biological Cybernetics, Max Planck Society, ou_2528697              
2Max Planck Institute for Biological Cybernetics, Max Planck Society, ou_1497794              
3Research Group Neural Population Imaging, Max Planck Institute for Biological Cybernetics, Max Planck Society, Spemannstrasse 38, 72076 Tübingen, DE, ou_1497807              

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 Abstract: While the neuronal basis of certain behaviors and sensory modalities can be studied under anesthesia or head-fixation, the full gamut of neural activity and its functions is accessible only in awake and unrestrained animals. We therefore developed a miniaturized, head mounted 2-photon microscope or “fiberscope” capable of resolving functional action-potential derived fluorescent signals from individual cortical cells. Using the fiberscope, we imaged spontaneous and stimulus-evoked activity from populations of layer 2/3 neurons and astrocytes in the visual cortex in both anesthetized and freely moving rats. The fiberscope weighs 5.5 g, and employs a custom-designed water immersion lens and a non-resonant fiber scanner leveraged by a piezo-element. Excitation light is delivered to the mobile animal through a single-mode optical fiber. Emitted light is collected through a plastic optical fiber before being split into a green channel, for the calcium-indicator Oregon Green BAPTA-1 (OGB1), and a red channel for the specific astrocyte marker sulforhodamin-101. Images acquired through the fiberscope were stable through a range of behaviors, even when the animal is moving vigorously (> 0.6 m/s). Using optical intrinsic imaging to locate the visual cortex binocular region, OGB1 was bulk loaded into cells within this region and remained viable over the course of the four hour experiments. Movement-based artifacts were small and could be successfully corrected offline using custom built software. Fiberscope imaging of neurons under anesthesia revealed robust stimulus responses and orientation selectivity. For imaging in freely-moving animals, the rat was allowed to run in a rectangular arena with one wall made of transparent Perspex behind which static visual stimuli were presented on a CRT monitor. Imaging sessions were run in darkness and recorded using infra-red videography. Transients could be observed in a subgroup of cells in response to activation of the visual stimulus. Further, movements of the animal which resulted in the stimulus sweeping through the animals visual field also evoked transients in a subgroup of the imaged cells. Thus, the miniaturized two-photon fiberscope can record spontaneous and stimulus-evoked Ca2+ transients in freely moving animals. We expect that the fiberscope will facilitate the study of neuronal population activity of during complex behavioral tasks.

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 Dates: 2009-10
 Publication Status: Published in print
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Title: 39th Annual Meeting of the Society for Neuroscience (Neuroscience 2009)
Place of Event: Chicago, IL, USA
Start-/End Date: 2009-10-17 - 2009-10-21

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Title: 39th Annual Meeting of the Society for Neuroscience (Neuroscience 2009)
Source Genre: Proceedings
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Pages: - Volume / Issue: - Sequence Number: 353.11 Start / End Page: - Identifier: -