Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Patch Clamp01:18

Patch Clamp

5.6K
Many fundamental cell functions such as muscle contraction and nerve transmission rely on the electrical signals produced by the movement of positively and negatively charged ions across the cell membrane. One competent method to record current flowing across the whole cell or single ion channel is the patch-clamp technique.
In this method, a glass micropipette containing electrolyte solution is tightly sealed against a small portion of the cell membrane. As a result, a patch of the cell...
5.6K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Targeting intracellular cholesterol imbalance rescues sarcomere-ER contact site signaling and ER remodeling in dilated cardiomyopathy.

Signal transduction and targeted therapy·2026
Same author

Resolving three GABA induced electrogenic events and GABA-coupling stoichiometry for Na<sup>+</sup> and Cl<sup>-</sup> in hGAT-1.

The FEBS journal·2026
Same author

Long-term outcomes of venous graft preservation in a rat autologous transplant model.

Scientific reports·2026
Same author

Transcriptomic analysis and high throughput functional characterization of human induced pluripotent stem cell derived sensory neurons.

Neurobiology of pain (Cambridge, Mass.)·2026
Same author

A modular method for high-throughput measurement of ion channel currents in cardiac myocytes.

Nature protocols·2026
Same author

Long-term outcomes of aortic grafts after 2-week storage in a rat model of orthotopic aortic transplantation.

Naunyn-Schmiedeberg's archives of pharmacology·2026

Related Experiment Video

Updated: Aug 29, 2025

Isolation of Human Atrial Myocytes for Simultaneous Measurements of Ca2+ Transients and Membrane Currents
10:53

Isolation of Human Atrial Myocytes for Simultaneous Measurements of Ca2+ Transients and Membrane Currents

Published on: July 3, 2013

20.5K

There is no F in APC: Using physiological fluoride-free solutions for high throughput automated patch clamp

Markus Rapedius1, Alison Obergrussberger1, Edward S A Humphries2

  • 1Nanion Technologies GmbH, Munich, Germany.

Frontiers in Molecular Neuroscience
|September 8, 2022
PubMed
Summary

This study presents a novel fluoride-free internal solution for automated patch clamp, achieving high GΩ seal success rates. This method enables reliable recordings of ion channels without fluoride

Keywords:
KCa3.1NaV1.5NaV1.7automated patch clampfluoridehERGion channelsphysiological solutions

More Related Videos

Hypothalamic Kisspeptin Neurons as a Target for Whole-Cell Patch-Clamp Recordings
09:39

Hypothalamic Kisspeptin Neurons as a Target for Whole-Cell Patch-Clamp Recordings

Published on: March 17, 2023

2.4K
Perforated Patch-clamp Recording of Mouse Olfactory Sensory Neurons in Intact Neuroepithelium: Functional Analysis of Neurons Expressing an Identified Odorant Receptor
10:16

Perforated Patch-clamp Recording of Mouse Olfactory Sensory Neurons in Intact Neuroepithelium: Functional Analysis of Neurons Expressing an Identified Odorant Receptor

Published on: July 13, 2015

26.8K

Related Experiment Videos

Last Updated: Aug 29, 2025

Isolation of Human Atrial Myocytes for Simultaneous Measurements of Ca2+ Transients and Membrane Currents
10:53

Isolation of Human Atrial Myocytes for Simultaneous Measurements of Ca2+ Transients and Membrane Currents

Published on: July 3, 2013

20.5K
Hypothalamic Kisspeptin Neurons as a Target for Whole-Cell Patch-Clamp Recordings
09:39

Hypothalamic Kisspeptin Neurons as a Target for Whole-Cell Patch-Clamp Recordings

Published on: March 17, 2023

2.4K
Perforated Patch-clamp Recording of Mouse Olfactory Sensory Neurons in Intact Neuroepithelium: Functional Analysis of Neurons Expressing an Identified Odorant Receptor
10:16

Perforated Patch-clamp Recording of Mouse Olfactory Sensory Neurons in Intact Neuroepithelium: Functional Analysis of Neurons Expressing an Identified Odorant Receptor

Published on: July 13, 2015

26.8K

Area of Science:

  • Electrophysiology
  • Ion Channel Research
  • Pharmacology

Background:

  • Fluoride-containing internal solutions are standard in patch clamp electrophysiology for enhancing seal resistance and recording duration.
  • Fluoride's presence can interfere with ion channel gating kinetics and intracellular signaling pathways, necessitating fluoride-free alternatives.
  • Automated patch clamp (APC) systems rely on internal fluoride to achieve high gigaseal (GΩ) formation rates, crucial for data quality.

Purpose of the Study:

  • To develop and validate a high-throughput, fluoride-free internal solution for automated patch clamp experiments.
  • To assess the efficacy of the fluoride-free approach in achieving high GΩ seal rates across various cell lines and expressed ion channels.
  • To provide a comprehensive comparison of fluoride-containing versus fluoride-free solutions in patch clamp electrophysiology.

Main Methods:

  • Development of a novel, high-throughput, fluoride-free internal recording solution.
  • Implementation on a 384-well based automated patch clamp system.
  • Validation using cells expressing human ether-à-go-go-related gene (hERG), sodium channel NaV1.5, NaV1.7, and potassium calcium-activated channel KCa3.1.

Main Results:

  • Achieved success rates exceeding 40% for gigaseal (GΩ) formation using the fluoride-free internal solution in a high-throughput APC system.
  • Successfully recorded from multiple ion channels including hERG, NaV1.5, NaV1.7, and KCa3.1 expressed in HEK and CHO cells.
  • Demonstrated the feasibility of performing patch clamp experiments without internal fluoride, preserving physiological channel behavior.

Conclusions:

  • The developed fluoride-free internal solution offers a viable and effective alternative for automated patch clamp recordings.
  • This approach overcomes the limitations of fluoride, such as altered channel gating and second messenger system interference.
  • The method is advantageous for studying ion channels where precise voltage-dependence and physiological intracellular conditions are critical.