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Cleaning patch-clamp pipettes for immediate reuse

Patch-clamp recording has enabled single-cell electrical, morphological and genetic studies at unparalleled resolution. Yet it remains a laborious and low-throughput technique, making it largely impractical for large-scale measurements such as cell type and connectivity characterization of neurons i...

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Autores principales: Kolb, I., Stoy, W. A., Rousseau, E. B., Moody, O. A., Jenkins, A., Forest, C. R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5057089/
https://www.ncbi.nlm.nih.gov/pubmed/27725751
http://dx.doi.org/10.1038/srep35001
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author Kolb, I.
Stoy, W. A.
Rousseau, E. B.
Moody, O. A.
Jenkins, A.
Forest, C. R.
author_facet Kolb, I.
Stoy, W. A.
Rousseau, E. B.
Moody, O. A.
Jenkins, A.
Forest, C. R.
author_sort Kolb, I.
collection PubMed
description Patch-clamp recording has enabled single-cell electrical, morphological and genetic studies at unparalleled resolution. Yet it remains a laborious and low-throughput technique, making it largely impractical for large-scale measurements such as cell type and connectivity characterization of neurons in the brain. Specifically, the technique is critically limited by the ubiquitous practice of manually replacing patch-clamp pipettes after each recording. To circumvent this limitation, we developed a simple, fast, and automated method for cleaning glass pipette electrodes that enables their reuse within one minute. By immersing pipette tips into Alconox, a commercially-available detergent, followed by rinsing, we were able to reuse pipettes 10 times with no degradation in signal fidelity, in experimental preparations ranging from human embryonic kidney cells to neurons in culture, slices, and in vivo. Undetectable trace amounts of Alconox remaining in the pipette after cleaning did not affect ion channel pharmacology. We demonstrate the utility of pipette cleaning by developing the first robot to perform sequential patch-clamp recordings in cell culture and in vivo without a human operator.
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spelling pubmed-50570892016-10-24 Cleaning patch-clamp pipettes for immediate reuse Kolb, I. Stoy, W. A. Rousseau, E. B. Moody, O. A. Jenkins, A. Forest, C. R. Sci Rep Article Patch-clamp recording has enabled single-cell electrical, morphological and genetic studies at unparalleled resolution. Yet it remains a laborious and low-throughput technique, making it largely impractical for large-scale measurements such as cell type and connectivity characterization of neurons in the brain. Specifically, the technique is critically limited by the ubiquitous practice of manually replacing patch-clamp pipettes after each recording. To circumvent this limitation, we developed a simple, fast, and automated method for cleaning glass pipette electrodes that enables their reuse within one minute. By immersing pipette tips into Alconox, a commercially-available detergent, followed by rinsing, we were able to reuse pipettes 10 times with no degradation in signal fidelity, in experimental preparations ranging from human embryonic kidney cells to neurons in culture, slices, and in vivo. Undetectable trace amounts of Alconox remaining in the pipette after cleaning did not affect ion channel pharmacology. We demonstrate the utility of pipette cleaning by developing the first robot to perform sequential patch-clamp recordings in cell culture and in vivo without a human operator. Nature Publishing Group 2016-10-11 /pmc/articles/PMC5057089/ /pubmed/27725751 http://dx.doi.org/10.1038/srep35001 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Kolb, I.
Stoy, W. A.
Rousseau, E. B.
Moody, O. A.
Jenkins, A.
Forest, C. R.
Cleaning patch-clamp pipettes for immediate reuse
title Cleaning patch-clamp pipettes for immediate reuse
title_full Cleaning patch-clamp pipettes for immediate reuse
title_fullStr Cleaning patch-clamp pipettes for immediate reuse
title_full_unstemmed Cleaning patch-clamp pipettes for immediate reuse
title_short Cleaning patch-clamp pipettes for immediate reuse
title_sort cleaning patch-clamp pipettes for immediate reuse
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5057089/
https://www.ncbi.nlm.nih.gov/pubmed/27725751
http://dx.doi.org/10.1038/srep35001
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