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In this protocol, whole cell patch-clamp recordings were made from neurons using an intracellular recording solution containing 50 µM Alexa Fluor® 633. Thus, only the recorded neurone will be filled with a fluorescent dye that will subsequently be visualized on the confocal microscope following the treatment of the slice with this protocol.
Remove the brain slice from the recording chamber and submerge overnight in 4% PFA (paraformaldehyde) in 0.1 M phosphate buffer pH 7.4.
Once coverslip has been applied, the sample is ready for visualization with the confocal microscope.
The video protocol was produced by Abcam in partnership with NeuroSolutions.