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dc.contributor.author
Vaglienti, María Victoria  
dc.contributor.author
Subirada Caldarone, Paula Virginia  
dc.contributor.author
Barcelona, Pablo Federico  
dc.contributor.author
Bonacci, Gustavo Roberto  
dc.contributor.author
Sanchez, Maria Cecilia  
dc.date.available
2023-11-06T14:55:00Z  
dc.date.issued
2022-05  
dc.identifier.citation
Vaglienti, María Victoria; Subirada Caldarone, Paula Virginia; Barcelona, Pablo Federico; Bonacci, Gustavo Roberto; Sanchez, Maria Cecilia; Quantification of Reactive Oxygen Species Using 2′,7′- Dichlorofluorescein Diacetate Probe and Flow-Cytometry in Müller Glial Cells; Journal of Visualized Experiments; Journal of Visualized Experiments; 2022; 183; 5-2022; 1-16  
dc.identifier.issn
1940-087X  
dc.identifier.uri
http://hdl.handle.net/11336/217168  
dc.description.abstract
The redox balance has an important role in maintaining cellular homeostasis. The increased generation of reactive oxygen species (ROS) promotes the modification of proteins, lipids, and DNA, which finally may lead to alteration in cellular function and cell death. Therefore, it is beneficial for cells to increase their antioxidant defense in response to detrimental insults, either by activating an antioxidant pathway like Keap1/Nrf2 or by improving redox scavengers (vitamins A, C, and E, β-carotene,and polyphenols, among others). Inflammation and oxidative stress are involved in the pathogenesis and progression of retinopathies, such as diabetic retinopathy (DR) and retinopathy of prematurity (ROP). Since Müller glial cells (MGCs) play a key role in the homeostasis of neural retinal tissue, they are considered an ideal model to study these cellular protective mechanisms. In this sense, quantifying ROS levels with a reproducible and simple method is essential to assess the contribution of pathways or molecules that participate in the antioxidant cell defense mechanism. In this article, we provide a complete description of the procedures required for the measurement of ROS with DCFH-DA probe and flow cytometry in MGCs. Key steps for flow cytometry data processing with the software are provided here, so the readers will be able to measure ROS levels (geometric means of FITC) and analyze fluorescence histograms. These tools are highly helpful to evaluate not only the increase in ROS after a cellular insult but also to study the antioxidant effect of certain molecules that can provide a protective effect on the cells.  
dc.format
application/pdf  
dc.language.iso
eng  
dc.publisher
Journal of Visualized Experiments  
dc.rights
info:eu-repo/semantics/restrictedAccess  
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/  
dc.subject
Reactive oxygen species (ROS)  
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Müller glial cells (MGCs)  
dc.subject
Diabetic retinopathy (DR)  
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Retinopathy of prematurity (ROP  
dc.subject.classification
Bioquímica y Biología Molecular  
dc.subject.classification
Ciencias Biológicas  
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CIENCIAS NATURALES Y EXACTAS  
dc.title
Quantification of Reactive Oxygen Species Using 2′,7′- Dichlorofluorescein Diacetate Probe and Flow-Cytometry in Müller Glial Cells  
dc.type
info:eu-repo/semantics/article  
dc.type
info:ar-repo/semantics/artículo  
dc.type
info:eu-repo/semantics/publishedVersion  
dc.date.updated
2023-10-30T17:59:33Z  
dc.journal.volume
2022  
dc.journal.number
183  
dc.journal.pagination
1-16  
dc.journal.pais
Estados Unidos  
dc.journal.ciudad
Cambridge  
dc.description.fil
Fil: Vaglienti, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina  
dc.description.fil
Fil: Subirada Caldarone, Paula Virginia. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina  
dc.description.fil
Fil: Barcelona, Pablo Federico. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina  
dc.description.fil
Fil: Bonacci, Gustavo Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina  
dc.description.fil
Fil: Sanchez, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Bioquímica Clínica; Argentina  
dc.journal.title
Journal of Visualized Experiments  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.3791/63337  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.jove.com/es/t/63337/quantification-reactive-oxygen-species-using-27-dichlorofluorescein