Chronic calcium imaging of neurons in the mouse visual cortex using a troponin C-based indicator.
This protocol describes the use of the genetically encoded troponin C-based calcium indicator TN-XXL to chronically monitor the functional properties of single neocortical neurons in the mouse visual cortex. A cranial window is implanted over the brain of a mouse expressing TN-XXL in pyramidal neurons of the cerebral cortex. Several days later, the visual cortex is mapped and photographed to facilitate repeated imaging of the same region using two-photon microscopy. Initial two-photon imaging may be done ∼2 wk after the window is implanted. We show the application of this technique for long-term in vivo imaging of stimulus response properties. Beyond providing functional information, long-term imaging of TN-XXL-labeled neurons also enables the simultaneous monitoring of structural properties down to the level of single dendritic spines.
Alexandre Ferrão
AF
Santos
Mark
M
Hübener
D000818:Animals; D002118:Calcium / Q000032:analysis*; D020013:Calcium Signaling; D007202:Indicators and Reagents / Q000032:analysis*; D051379:Mice; D009474:Neurons / Q000737:chemistry / Q000502:physiology*; D061848:Optical Imaging / Q000379:methods*; D013997:Time Factors; D019209:Troponin C / Q000032:analysis*; D014793:Visual Cortex / Q000737:chemistry / Q000502:physiology*
D007202:Indicators and Reagents; D019209:Troponin C; D002118:Calcium