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

Fritjof Helmchen, Michale S. Fee, David W. Tank, Winfried Denk

Research output: Contribution to journalArticle

468 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)903-912
Number of pages10
JournalNeuron
Volume31
Issue number6
DOIs
StatePublished - Sep 27 2001
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

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