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. 2017 Jul 13;7:5318. doi: 10.1038/s41598-017-05607-3

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Patch clamp-assisted single neuron lipidomics. (a) Mouse brain slices are prepared using a vibratome and transferred to the perfusion chamber of a standard electrophysiology setup. A target neuron is visually identified and patched in the cell-attached configuration using a standard manipulator. A patched granule cell of the dentate gyrus and the attached pipette are shown in pseudocolor. (b) The neuronal soma is carefully pulled away from the slice while maintaining the giga-ohm seal. Visual inspection is used to confirm that the neuron is free of extraneous contaminating material such as glial cells; calibration bar: 10 μm (c) The patched neuron is aspirated into the pipette tip and transferred to a sterile vial kept in dry ice. Stimulated cells are collected from the slice within 3 min of stimulation. Samples are subjected to single-phase isopropanol extraction and lipids are analyzed by nanoflow liquid chromatography/high-resolution time-of-flight mass spectrometry (nLC-MS).