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Author Notes:

Correspondence and requests for materials should be addressed to C.R.F. (email: cforest@gatech.edu).

I.K., W.A.S. and C.R.F. conceived the project.

I.K. and O.A.M. performed in vitro experiments.

W.A.S. performed in vivo experiments.

E.B.R. performed SEM imaging.

Experimental work was supervised by A.J. and C.R.F.

The manuscript was written by I.K. and C.R.F. with contributions from all authors.

We thank The Georgia Tech Bioanalytical Mass Spectrometry Facility for MS analysis and Thomas Capocasale for SEM imaging. We thank the Allen Institute for Brain Science for hosting us and our collaborators there for continued helpful discussions.

The authors declare no competing financial interests.

Subjects:

Research Funding:

IK acknowledges the NIH Computational Neuroscience Training grant (DA032466-02) and Georgia Tech Neural Engineering Center Seed Grant.

CRF acknowledges NIH grants (1-U01-MH106027-01, 1-R01-EY023173, 5-R44-NS083108-03), Georgia Tech Fund for Innovation in Research and Education (GT-FIRE), support from Georgia Tech through the Institute for Bioengineering and Biosciences Junior Faculty Award, Technology Fee Fund, Invention Studio, and the George W. Woodruff School of Mechanical Engineering.

EBR acknowledges the National Science Foundation grant (ECCS-0335765).

Keywords:

  • Science & Technology
  • Multidisciplinary Sciences
  • NEURONS IN-VIVO
  • SYNAPTIC-TRANSMISSION
  • RECEPTOR FUNCTION
  • ION CHANNELS
  • BRAIN
  • RECORDINGS
  • MEMBRANE
  • SYSTEM
  • CORTEX
  • Cellular neuroscience
  • Patch clamp

Cleaning patch-clamp pipettes for immediate reuse

Tools:

Journal Title:

Scientific Reports

Volume:

Volume 6

Publisher:

, Pages 35001-35001

Type of Work:

Article | Final Publisher PDF

Abstract:

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.

Copyright information:

© The Author(s) 2016.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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