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JoVE Journal Neuroscience
Quantification of Reactive Oxygen Species Using 2′,7′-Dichlorofluorescein Diacetate Probe and Flow-Cytometry in Müller Glial Cells
 

Quantification of Reactive Oxygen Species Using 2′,7′-Dichlorofluorescein Diacetate Probe and Flow-Cytometry in Müller Glial Cells

Article DOI: 10.3791/63337-v • 14:25 min • May 13th, 2022
May 13th, 2022 •
1,2, 1,3, 1,2, 1,2, 1,2
1Departamento de Bioquímica Clínica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, 2Centro de Investigaciones en Bioquímica Clínica e Inmunología (CIBICI), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), 3Instituto de Investigación Médica Mercedes y Martín Ferreyra (INIMEC), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)

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요약
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Here, we propose a systematized, accessible, and reproducible protocol to detect cellular reactive oxygen species (ROS) using 2′,7′-dichlorofluorescein diacetate probe (DCFH-DA) in Müller glial cells (MGCs). This method quantifies total cellular ROS levels with a flow cytometer. This protocol is very easy to use, suitable, and reproducible.

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Keywords: Reactive Oxygen Species ROS 2',7'-dichlorofluorescein Diacetate DCFDA Flow Cytometry Oxidative Stress Cell Membrane Esterases Dichlorofluorescein Antioxidant ROS Inducer DMEM PBS Centrifugation FACS Buffer
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