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Biologie du développement
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Evaluation of Intracellular Location of Reactive Oxygen Species in Solea Senegalensis Spermatozoa
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Biologie du développement
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Journal JoVE Biologie du développement
Evaluation of Intracellular Location of Reactive Oxygen Species in Solea Senegalensis Spermatozoa

Evaluation of Intracellular Location of Reactive Oxygen Species in Solea Senegalensis Spermatozoa

6,134 Views

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10:24 min

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March 11, 2018

DOI:

10.3791/55323-v

DOI:
10.3791/55323-v

•

10:24 min
March 11, 2018

•

4 Views
,
1Department of Molecular Biology,University of León, 2INDEGSAL,University of León, 3Planta de Cultivos el Bocal,Spanish Institute of Oceanography (IEO)

Summary

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This protocol describes a detailed methodology for detecting H2O2 localization within Solea senegalensis spermatozoa using a sensitive fluorochrome DCFH-DA for ROS, a live mitochondria stain for mitochondria, and DAPI for nuclei visualization, respectively. The protocol is designed to be performed within 2 h with either fresh or thawed spermatozoa.

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Reactive Oxygen SpeciesSpermatozoaSolea SenegalensisConfocal MicroscopyFluorochromesMitochondriaDAPI2′7′-dichlorofluorescein DiacetateOxidative StressLipid PeroxidationApoptosisCryopreservation

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Cite this Article

Valcarce, D. G., Robles, V. Evaluation of Intracellular Location of Reactive Oxygen Species in Solea Senegalensis Spermatozoa. J. Vis. Exp. (133), e55323, doi:10.3791/55323 (2018).

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