Parallel two-photon imaging and electrophysiological recordings provide different aspects to study the neuronal cell and network activity:
- the electrical signals show local changes on the studied area
- and the changes in fluorescence reveal the calcium currents at multiple sites of the cell or network
Patch-clamp recording and Ca2+ imaging in OGB-1 and Alexa-594 filled hippocampal neuron using Femto2D-Galvo. The two parallel signals are time-aligned and co-analyzed by MES software.
The electrical signals can be digitalized directly by the microscope control electronics or by the separated external digitizer and software imported later. In both cases the analysis is performed by the MES control software.
Dendritic patch clamp recording
- automatic software module moves the tip of the pipette to the selected dendritic location
- guide points can be positioned by the user according to the scanned transmitted or two-photon images
- the recording pipettes are moved in the tissue only diagonally in order to minimize tissue damage
- pipette movement and distance to target location can be monitored real-time
- only a simple calibration step is required before using a new recording pipette
- the final steps of dendritic targeting can be controlled both manually and automatically
- real-time transmitted infrared and two photon images can be shown separately or in overlay
Simultaneous real-time visualization of the patch pipette and the dendrite in order to create dendritic patch-clamp recording from thin dendrites (tIR: transmitted infrared).