TY - JOUR
T1 - A miniature head-mounted two-photon microscope
T2 - High-resolution brain imaging in freely moving animals
AU - Helmchen, Fritjof
AU - Fee, Michale S.
AU - Tank, David W.
AU - Denk, Winfried
N1 - Funding Information:
We thank G. Buzsaki for discussions; and M. Brecht and D.J. Waters for critically reading the manuscript. This work was supported by Lucent Technologies and the Whitehall Foundation. F.H. acknowledges support by a postdoctoral fellowship from the Human Frontier Science Program.
PY - 2001/9/27
Y1 - 2001/9/27
N2 - 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.
AB - 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.
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U2 - 10.1016/S0896-6273(01)00421-4
DO - 10.1016/S0896-6273(01)00421-4
M3 - Article
C2 - 11580892
AN - SCOPUS:0035959949
SN - 0896-6273
VL - 31
SP - 903
EP - 912
JO - Neuron
JF - Neuron
IS - 6
ER -