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.
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Acknowledgements
This work was supported in part by the Funding Program for Next Generation World-Leading Researchers (LS023).
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T.S. collected experimental data and carried out the data analysis. T.S. and Y.I. wrote the manuscript. N.M. supervised the project and provided feedback on the manuscript.
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Supplementary information
Supplementary Video 1
Targeted axon-attached recording from an axon of a hippocampal pyramidal cell. (AVI 8228 kb)
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Sasaki, T., Matsuki, N. & Ikegaya, Y. Targeted axon-attached recording with fluorescent patch-clamp pipettes in brain slices. Nat Protoc 7, 1228–1234 (2012). https://doi.org/10.1038/nprot.2012.061
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DOI: https://doi.org/10.1038/nprot.2012.061
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