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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.

Author information

1
Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan. t.sasaki.0224@gmail.com

Abstract

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.

PMID:
22653161
DOI:
10.1038/nprot.2012.061
[Indexed for MEDLINE]

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