title: Cleaning patch-clamp pipettes for immediate reuse.
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.
authors.affiliation: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332 USA.
authors.forename: I
authors.initials: I
authors.lastname: Kolb
authors.affiliation: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332 USA.
authors.forename: W A
authors.initials: WA
authors.lastname: Stoy
authors.affiliation: Colleges of Nanoscale Science and Engineering, SUNY Polytechnic Institute, Albany, NY, 12203 USA.
authors.forename: E B
authors.initials: EB
authors.lastname: Rousseau
authors.affiliation: Neuroscience Graduate Program, GDBBS, Emory University, Atlanta, GA, 30322 USA.
authors.forename: O A
authors.initials: OA
authors.lastname: Moody
authors.affiliation: Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, 30322 USA.|Department of Anesthesiology, Emory University, Atlanta, GA, 30322 USA.
authors.forename: A
authors.initials: A
authors.lastname: Jenkins
authors.affiliation: George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta GA, 30332 USA.
authors.forename: C R
authors.initials: CR
authors.lastname: Forest
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003902:Detergents; D005898:Glass; D057809:HEK293 Cells; D006801:Humans; D008839:Microelectrodes; D009474:Neurons / Q000166:cytology*; D018408:Patch-Clamp Techniques / Q000295:instrumentation* / Q000379:methods; D051381:Rats
chemical_list: D003902:Detergents