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A miniature head−mounted two−photon microscope: High−resolution brain imaging in freely moving animals

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Helmchen,  Fritjof
Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society;
In Vivo Microscopy of Cortical Dynamics, Max Planck Institute for Medical Research, Max Planck Society;
Cortical Two Photon Imaging, Max Planck Institute for Medical Research, Max Planck Society;

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Denk,  Winfried
Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society;

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Citation

Helmchen, F., Fee, M. S., Tank, D. W., & Denk, W. (2001). A miniature head−mounted two−photon microscope: High−resolution brain imaging in freely moving animals. Neuron, 31, 903-912. doi:10.1016/S0896-6273(01)00421-4.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0019-A043-7
Abstract
Two−photon microscopy has enabled anatomical and functional fluorescence imaging in the intact brain of rats. Here, we extend two−photon imaging from anesthetized, head−stabilized to awake, freely moving animals by using a miniaturized head−mounted microscope. Excitation light is conducted to the microscope in a single−mode optical fiber, and images are scanned using vibrations of the fiber tip. Microscope performance was first characterized in the neocortex of anesthetized rats. We readily obtained images of vasculature filled with fluorescently labeled blood and of layer 2/3 pyramidal neurons filled with a calcium indicator. Capillary blood flow and dendritic calcium transients were measured with high time resolution using line scans. In awake, freely moving rats, stable imaging was possible except during sudden head movements