In vivo cell-attached recordings were performed using glass pipettes (∼5-7 MΩ) filled with solution containing the following (in mM): 125 NaCl, 5 KCl, 10 glucose, 10 HEPES, 2 CaCl2, 2 MgSO4, and 0.1 Alexa Fluor 594; pH 7.4). Signals were amplified using an AxoPatch 200B amplifier (Molecular Devices), filtered at 5 kHz, and digitized at 10 kHz. Spikes were recorded using current clamp mode. The frame trigger pulses of ScanImage 4.0 were also recorded and used offline to synchronize individual frames to electrophysiological recordings. After establishment of a low-resistance seal (15-50 MΩ), the orientation, spatial and temporal frequency of the stimuli was quickly optimized for individual neurons using recorded spikes. The optimal grating stimulus was repeated at a reduced contrast to maintain a moderate spiking rate.
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In vivo Cell-Attached Electrophysiology
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Other organizations : Janelia Research Campus, Helix (United States), Howard Hughes Medical Institute, Champalimaud Foundation
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Variable analysis
independent variables
- Orientation of the stimulus
- Spatial frequency of the stimulus
- Temporal frequency of the stimulus
- Spiking rate of neurons
- Glass pipette resistance (∼5-7 MΩ)
- Composition of the pipette solution (125 mM NaCl, 5 mM KCl, 10 mM glucose, 10 mM HEPES, 2 mM CaCl2, 2 mM MgSO4, 0.1 mM Alexa Fluor 594; pH 7.4)
- Amplifier settings (AxoPatch 200B, filtered at 5 kHz, digitized at 10 kHz)
- Seal resistance (15-50 MΩ)
- Positive control: Not applicable
- Negative control: Not applicable
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