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Nat Protoc. 2012 May 31;7(6):1228-34. doi: 10.1038/nprot.2012.061.

Targeted axon-attached recording with fluorescent patch-clamp pipettes in brain slices.

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Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan.


Understanding the physiology of axons in the central nervous system requires experimental access to intact axons. This protocol describes how to perform cell-attached recordings from narrow axon fibers (ϕ <1 μm) in acute and cultured brain slice preparations (with a success rate of ∼50%). By using fluorophore-coated glass pipettes and Nipkow disk confocal microscopy, fluorescently labeled axons can be visually targeted under online optical control. In the cell-attached configuration, axonal action potentials are extracellularly recorded as unit-like, sharp negative currents. The axon morphology labeling and cell-attached recordings of axons can be completed within 1-2 h. The recordings are stable for at least 30 min.

[Indexed for MEDLINE]

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