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Evaluation of Intracellular Location of Reactive Oxygen Species in Solea Senegalensis Spermatozoa
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Entwicklungsbiologie
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JoVE Journal Entwicklungsbiologie
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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