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Quantification of Reactive Oxygen Species Using 2′,7′-Dichlorofluorescein Diacetate Probe and Flow-Cytometry in Müller Glial Cells
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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)
Capitoli
- 00:00Introduction
- 01:37Performing the Assay
- 04:21Cells Preparaion for Flow Cytometry
- 06:57Data Acquisition in a Flow Cytometer
- 09:00Analysis of the Acquired Data
- 12:07Representative Results
- 13:37Conclusion
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.
