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Related Concept Videos

Patch Clamp01:18

Patch Clamp

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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.
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Amperometry: Overview01:10

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Amperometry is a technique commonly used to measure the concentration of specific analytes in a solution by monitoring the electric current generated during an electrochemical reaction. It involves applying a constant potential between a working electrode and a reference electrode to measure the resulting current, which is proportional to the concentration of the analyte. The Clark oxygen electrode operates based on this principle of amperometry. It consists of a cathode and an anode enclosed...
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Membrane electrodes, also known as p-ion electrodes, use membranes that selectively interact with free analyte ions, generating a potential difference across the membrane. The resulting membrane potential, known as the asymmetry potential, is not zero even when analyte concentrations on both sides of the membrane are equal. The membrane's response is typically not selective to a single analyte but proportional to the concentration of all ions in the sample solution capable of interacting at...
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Interfacial electrochemical methods focus on the phenomena occurring at the boundary between an electrode and a solution, as opposed to bulk methods that concentrate on the solution's overall properties. These interfacial methods are classified as either static or dynamic based on the presence of a nonzero current in the electrochemical cell and the consistency of analyte concentrations. Static methods, such as potentiometry, measure the cell's potential without any significant current...
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Single Cell Measurement of Dopamine Release with Simultaneous Voltage-clamp and Amperometry
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Patch Amperometry and Intracellular Patch Electrochemistry.

Eugene V Mosharov1, Manfred Lindau2

  • 1Departments of Psychiatry and Neurology, Columbia University Medical Center, New York State Psychiatric Institute, New York, NY, USA. johnmosh@yahoo.com.

Methods in Molecular Biology (Clifton, N.J.)
|October 7, 2022
PubMed
Summary
This summary is machine-generated.

Patch amperometry and intracellular patch electrochemistry offer precise measurements of catecholamines. These electrochemical techniques provide detailed insights into exocytosis and cytosolic transmitter concentrations.

Keywords:
CatecholamineCyclic voltammetryCytosolicElectrochemistryExocytosisFusion poreIPEPatch amperometrySynaptic vesicle

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Area of Science:

  • Neuroscience
  • Electrochemistry
  • Cell Biology

Background:

  • Patch clamp electrophysiology is a powerful tool for studying cellular function.
  • Electrochemical detection offers high sensitivity and specificity for neurotransmitter analysis.

Purpose of the Study:

  • To provide detailed instructions for setting up and performing patch amperometry (PA) and intracellular patch electrochemistry (IPE).
  • To enable researchers to analyze catecholamine release and cytosolic concentrations with high resolution.

Main Methods:

  • Utilizing a carbon fiber electrode (CFE) housed within a patch pipette for electrochemical detection.
  • Employing cell-attached or excised inside-out configurations for PA and whole-cell configuration for IPE.
  • Applying amperometric and cyclic voltammetric detection modes for catecholamine analysis.

Main Results:

  • PA allows high-resolution patch capacitance measurements and simultaneous detection of released catecholamines.
  • IPE, in amperometric mode, detects total catechols, while cyclic voltammetry offers selective quantification of catecholamines and metabolites.
  • Both methods provide precise data on vesicle size, quantal content, fusion pore dynamics, and cytosolic transmitter concentrations.

Conclusions:

  • PA and IPE are versatile techniques for studying catecholaminergic signaling at the single-cell level.
  • Detailed protocols facilitate the application of these methods for investigating neurotransmitter release and metabolism.