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

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


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials
  • Animals
  • Axons / physiology*
  • Brain*
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / metabolism
  • Image Processing, Computer-Assisted
  • In Vitro Techniques
  • Mice
  • Mice, Inbred ICR
  • Microscopy, Confocal
  • Patch-Clamp Techniques / instrumentation*
  • Patch-Clamp Techniques / methods*
  • Rats
  • Rats, Wistar
  • Software


  • Fluorescent Dyes