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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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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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ROS Quantification Reactive Oxygen Species 2'7'dichlorofluorescein Diacetate Probe Flow Cytometry Oxidative Stress Measurement Redox State Measurement Intracellular Esterases Cleavage Cell Membrane Permeability Fluorescent Product Dichlorofluorescein Detection Of ROS Confocal Microscopy Quantification Of ROS By Flow Cytometry Antioxidant Treatment ROS Inducer Treatment
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