[en] The patch-clamp technique has revolutionized neurophysiology by allowing to study single neuronal excitability, synaptic connectivity, morphology, and the transcriptomic profile. However, the throughput in recordings is limited because of the manual replacement of patch-pipettes after each attempt which are often also unsuccessful. This has been overcome by automated cleaning the tips in detergent solutions, allowing to reuse the pipette for further recordings. Here, we developed a novel method of automated cleaning by sonicating the tips within the bath solution wherein the cells are placed, reducing the risk of contaminating the bath solution or internal solution of the recording pipette by any detergent and avoiding the necessity of a separate chamber for cleaning. We showed that the patch-pipettes can be used consecutively at least ten times and that the cleaning process does not negatively impact neither the brain slices nor other patched neurons. This method, combined with automated patch-clamp, highly improves the throughput for single and especially multiple recordings.
Disciplines :
Engineering, computing & technology: Multidisciplinary, general & others
Author, co-author :
Jehasse, Kevin ; Université de Liège - ULiège > Département d'électricité, électronique et informatique (Institut Montefiore) > Systèmes et modélisation
Jouhanneau, Jean-Sébastien; Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany ; Neuroscience Research Center, Charité-Universitätsmedizin Berlin, Berlin, Germany
Wetz, Sophie; Systems Neurophysiology, Institute of Biology II, RWTH-Aachen University, Aachen, Germany ; Research Training Group 2610 InnoRetVision, RWTH-Aachen University, Aachen, Germany
Schwedt, Alexander; Central Facility for Electron Microscopy, RWTH-Aachen University, Aachen, Germany
Poulet, James F A; Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany ; Neuroscience Research Center, Charité-Universitätsmedizin Berlin, Berlin, Germany
Kampa, Björn M; Systems Neurophysiology, Institute of Biology II, RWTH-Aachen University, Aachen, Germany. kampa@brain.rwth-aachen.de ; JARA BRAIN, Institute of Neuroscience and Medicine (INM-10), Forschungszentrum Jülich, Jülich, Germany. kampa@brain.rwth-aachen.de ; Research Training Group 2610 InnoRetVision, RWTH-Aachen University, Aachen, Germany. kampa@brain.rwth-aachen.de
Language :
English
Title :
Immediate reuse of patch-clamp pipettes after ultrasonic cleaning.
Luigs & Neumann GmbH [DE] DFG - Deutsche Forschungsgemeinschaft [DE] NINDS - National Institute of Neurological Disorders and Stroke [US-MD] ERC - European Research Council [BE] HGF - Helmholtz Association of German Research Centres [DE] RWTH Aachen University [DE]
Funding text :
This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) Project
number 424556709-GRK 2610, the European Research Council (ERC-2015-CoG-682422, J.F.A.P.), the Deutsche
Forschungsgemeinschaft (FOR 2143, J.F.A.P.; SFB 1315, J.F.A.P.), the Helmholtz Society (J.F.A.P.), the National
Institute Of Neurological Disorders and Stroke of the National Institutes of Health (R01NS123711, subaward
number 1090705-458441, J.F.A.P., J-S.J; note that the content of this manuscript is solely the responsibility of the
authors and does not necessarily represent the official views of the National Institutes of Health) and by Luigs
& Neumann GmbH. We thank members of the laboratory of J.F.A.P. for constructive comments on the manuscript,
S. Steinfelder for help with administrative and technical aspects. We also thank the technical staff from
the Central Facility for Electron Microscopy at RWTH-Aachen University and the staff from Luigs & Neumann
GmbH involved in that project for their help.
Bezanilla, F., Rojas, E. & Taylor, R. E. Sodium and potassium conductance during a membrane action potential. J. Physiol. 211, 729–751 (1970). DOI: 10.1113/jphysiol.1970.sp009301
de Hass, V. & Vogel, W. Sodium and potassium currents recorded during an action potential. Eur. Biophys. J. 17, 49–51 (1989). DOI: 10.1007/BF00257145
Spruston, N. & Johnston, D. Perforated patch-clamp analysis of the passive membrane properties of three classes of hippocampal neurons. J. Neurophysiol. 67, 508–529 (1992). DOI: 10.1152/jn.1992.67.3.508
Petersen, C. C. H. Whole-cell recording of neuronal membrane potential during behavior. Neuron 95, 1266–1281 (2017). DOI: 10.1016/j.neuron.2017.06.049
Chen, X., Leischner, U., Rochefort, N. L., Nelken, I. & Konnerth, A. Functional mapping of single spines in cortical neurons in vivo. Nature 475, 501–505 (2011). DOI: 10.1038/nature10193
Edwards, F. A., Konnerth, A. & Sakmann, B. Quantal analysis of inhibitory synaptic transmission in the dentate gyrus of rat hippocampal slices: A patch-clamp study. J. Physiol. 430, 213–249 (1990). DOI: 10.1113/jphysiol.1990.sp018289
Bi, G. Q. & Poo, M. M. Synaptic modifications in cultured hippocampal neurons: Dependence on spike timing, synaptic strength, and postsynaptic cell type. J. Neurosci. 18, 10464–10472 (1998). DOI: 10.1523/JNEUROSCI.18-24-10464.1998
Kampa, B. M., Letzkus, J. J. & Stuart, G. J. Dendritic mechanisms controlling spike-timing-dependent synaptic plasticity. Trends Neurosci. 30, 456–463 (2007). DOI: 10.1016/j.tins.2007.06.010
Kampa, B. M., Letzkus, J. J. & Stuart, G. J. Cortical feed-forward networks for binding different streams of sensory information. Nat. Neurosci. 9, 1472–1473 (2006). DOI: 10.1038/nn1798
Taof, C., Zhangf, G., Xiong, Y. & Zhou, Y. Functional dissection of synaptic circuits: In vivo patch-clamp recording in neuroscience. Front. Neural Circuits 9, 31 (2015).
Stuart, G. J. & Sakmann, B. Active propagation of somatic action potentials into neocortical pyramidal cell dendrites. Nature 367, 69–72 (1994). DOI: 10.1038/367069a0
Shu, Y., Hasenstaub, A., Duque, A., Yu, Y. & McCormick, D. A. Modulation of intracortical synaptic potentials by presynaptic somatic membrane potential. Nature 441, 761–765 (2006). DOI: 10.1038/nature04720
Seutin, V. & Engel, D. Differences in Na+ conductance density and Na+ channel functional properties between dopamine and GABA neurons of the rat substantia nigra. J. Neurophysiol. 103, 3099–3114 (2010). DOI: 10.1152/jn.00513.2009
Kole, M. H. P. et al. Action potential generation requires a high sodium channel density in the axon initial segment. Nat. Neurosci. 11, 178–186 (2008). DOI: 10.1038/nn2040
Lee, B. R. et al. Scaled, high fidelity electrophysiological, morphological, and transcriptomic cell characterization. Elife 10, 65482 (2021). DOI: 10.7554/eLife.65482
Cadwell, C. R. et al. Electrophysiological, transcriptomic and morphologic profiling of single neurons using Patch-seq. Nat. Biotechnol. 34, 199–203 (2016). DOI: 10.1038/nbt.3445
Li, L. et al. A robot for high yield electrophysiology and morphology of single neurons in vivo. Nat. Commun. 8, 1–10 (2017).
Wang, G. et al. An optogenetics- and imaging-assisted simultaneous multiple patch-clamp recording system for decoding complex neural circuits. Nat. Protoc. 10, 397–412 (2015). DOI: 10.1038/nprot.2015.019
Koos, K. et al. Automatic deep learning-driven label-free image-guided patch clamp system. Nat. Commun. 12, 1–11 (2021). DOI: 10.1038/s41467-021-21291-4
Neher, E. Ion channels for communication between and within cells. Neuron 8, 605–612 (1992). DOI: 10.1016/0896-6273(92)90083-P
Kolb, I. et al. Cleaning patch-clamp pipettes for immediate reuse. Sci. Rep. 6, 1–10 (2016). DOI: 10.1038/srep35001
Landry, C. R. et al. Method for rapid enzymatic cleaning for reuse of patch clamp pipettes: Increasing throughput by eliminating manual pipette replacement between patch clamp attempts. Bio Protoc. 11, 4085 (2021). DOI: 10.21769/BioProtoc.4085
Kolb, I. et al. PatcherBot: A single-cell electrophysiology robot for adherent cells and brain slices. J. Neural Eng. 16, 046003 (2019). DOI: 10.1088/1741-2552/ab1834
Holst, G. L. et al. Autonomous patch-clamp robot for functional characterization of neurons in vivo: Development and application to mouse visual cortex. J. Neurophysiol. 121, 2341–2357 (2019). DOI: 10.1152/jn.00738.2018
Kodandaramaiah, S. B. et al. Multi-neuron intracellular recording in vivo via interacting autopatching robots. Elife 7, 24656 (2018). DOI: 10.7554/eLife.24656
Davie, J. T. et al. Dendritic patch-clamp recording. Nat. Protoc. 1, 1235–1247 (2006). DOI: 10.1038/nprot.2006.164
Kao, L., Abuladze, N., Shao, X. M., McKeegan, K. & Kurtz, I. A new technique for multiple re-use of planar patch clamp chips. J. Neurosci. Methods 208, 205–210 (2012). DOI: 10.1016/j.jneumeth.2012.05.002
Peng, Y. et al. High-throughput microcircuit analysis of individual human brains through next-generation multineuron patch-clamp. Elife 8, 48178 (2019). DOI: 10.7554/eLife.48178
Halfmann, C., Rüland, T., Müller, F., Jehasse, K. & Kampa, B. M. Electrophysiological properties of layer 2/3 pyramidal neurons in the primary visual cortex of a retinitis pigmentosa mouse model (rd10). Front. Cell Neurosci. 17, 1258773 (2023). DOI: 10.3389/fncel.2023.1258773
Ciganok-Hückels, N. et al. Postnatal development of electrophysiological and morphological properties in layer 2/3 and layer 5 pyramidal neurons in the mouse primary visual cortex. Cereb. Cortex 33, 5875–5884 (2023). DOI: 10.1093/cercor/bhac467
Kitamura, K., Judkewitz, B., Kano, M., Denk, W. & Häusser, M. Targeted patch-clamp recordings and single-cell electroporation of unlabeled neurons in vivo. Nat. Methods 5, 61–67 (2008). DOI: 10.1038/nmeth1150
Jouhanneau, J. S. & Poulet, J. F. A. Multiple two-photon targeted whole-cell patch-clamp recordings from monosynaptically connected neurons in vivo. Front. Synaptic Neurosci. 11, 444417 (2019). DOI: 10.3389/fnsyn.2019.00015
