Journal
NEURON
Volume 56, Issue 1, Pages 43-57Publisher
CELL PRESS
DOI: 10.1016/j.neuron.2007.08.003
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Funding
- NIDA NIH HHS [R90 DA023419-01, R90 DA023419] Funding Source: Medline
- NIGMS NIH HHS [T32 GM007388, T32 GM007388-31] Funding Source: Medline
- NIMH NIH HHS [R01 MH083686, K25 MH067876-01] Funding Source: Medline
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We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball's upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their heads remain motionless. Image sequences demonstrate that running-associated brain motion is limited to similar to 2-5 lam. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom-designed Hidden-Markov-Model-based motion correction algorithm useful for postprocessing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion-induced false positive error rate of < 5%.
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