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Related Concept Videos

Patch Clamp01:18

Patch Clamp

7.3K
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...
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Transport Number01:31

Transport Number

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The transport number is the fraction of the total current carried by an ion in an electrolyte solution. It is defined as the ratio of the current carried by a specific ion to the total current flowing through the solution. The transport number, t, is central to understanding ionic mobility, which describes how fast an ion moves under the influence of an electric field. This link connects the physical behavior of ions in solution to the chemical processes that occur during electrochemical...
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Related Experiment Video

Updated: Mar 18, 2026

Measurement of Ion Concentration in the Unstirred Boundary Layer with Open Patch-Clamp Pipette: Implications in Control of Ion Channels by Fluid Flow
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Measurement of Ion Concentration in the Unstirred Boundary Layer with Open Patch-Clamp Pipette: Implications in Control of Ion Channels by Fluid Flow

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Patch-clamp fluorometry-based channel counting to determine HCN channel conductance.

Chang Liu1, Changan Xie2, Khade Grant2

  • 1Department of Physiology and Biophysics, School of Medicine, Virginia Commonwealth University, Richmond, VA 23298 School of Medicine, Nankai University, Tianjin 300071, China.

The Journal of General Physiology
|June 30, 2016
PubMed
Summary

We developed a new patch-clamp fluorometry (PCF) method to count ion channels on cell membranes. This technique accurately quantifies hyperpolarization-activated and cyclic nucleotide-regulated (HCN) channels, enabling precise biophysical measurements.

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Last Updated: Mar 18, 2026

Measurement of Ion Concentration in the Unstirred Boundary Layer with Open Patch-Clamp Pipette: Implications in Control of Ion Channels by Fluid Flow
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Area of Science:

  • Biophysics
  • Cell Biology
  • Molecular Physiology

Background:

  • Accurate ion channel counting is crucial for understanding cell membrane biophysics.
  • Traditional methods like radioactive labeling and noise analysis have limitations.
  • Hyperpolarization-activated and cyclic nucleotide-regulated (HCN) channels play vital roles in cellular function.

Purpose of the Study:

  • To develop and validate a novel method for counting ion channels on cell membranes.
  • To apply this method to quantify hyperpolarization-activated and cyclic nucleotide-regulated (HCN) channels.
  • To determine the single-channel conductance of HCN1 and HCN2 channels.

Main Methods:

  • Development of patch-clamp fluorometry (PCF) for simultaneous electrical and optical recordings.
  • Application of PCF to EGFP-tagged HCN channels.
  • Calibration using homologous cyclic nucleotide-gated (CNG) channels to correlate fluorescence intensity with channel number.

Main Results:

  • Established a reliable relationship between macroscopic fluorescence intensity and total channel number.
  • Quantified the total number of HCN channels on cell membranes.
  • Determined the single-channel conductance for HCN1 (0.46 pS) and HCN2 (1.71 pS).

Conclusions:

  • Patch-clamp fluorometry (PCF) offers a robust alternative for ion channel counting.
  • The PCF method overcomes limitations of traditional electrophysiological techniques for small conductance channels.
  • This technique has broad applicability for biophysical studies of various membrane proteins.