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Single Cell Measurement of Dopamine Release with Simultaneous Voltage-clamp and Amperometry
Published on: November 21, 2012
Quantal Release Analysis of Electrochemically Active Molecules Using Single-Cell Amperometry
José David Machado1, Pablo Montenegro2, Natalia Domínguez3
1Dpto. Medicina Física y Farmacología, Facultad de Ciencias de la Salud, Medicina, Universidad de La Laguna, La Laguna, Tenerife, Spain. david.machado@ull.es.
Abstract:
Single-cell amperometry is a powerful technique that permits the detection of electrochemically active transmitters, such as catecholamines, histamine, or serotonin, released by exocytosis from secretory cells.Amperometry has two main characteristics that make it ideal for the study of exocytosis at the single-cell level with single-vesicle resolution quantal release. (i) It is noninvasive. The carbon fiber microelectrode can be carefully positioned on plasma membrane of a single cell, allowing the detection of the oxidation current of the secreted molecules. (ii) High temporal resolution and sensitivity. Exocytosis can be monitored with a real-time resolution that allows the determination of the kinetics release with an attomol detection sensitivity, which ensures an accurate calculation of the amount of transmitter released.Here, we compile some recommendations and advices to perform amperometry quantal analysis.
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