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Flow Cytometry and Confocal Microscopy for ROS Evaluation in Fish and Human Spermatozoa

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Reactive Oxygen Species

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2202))

Abstract

Reactive oxygen species (ROS) could have a negative impact on sperm cellular function and viability. This chapter describes a protocol for oxidative stress evaluation using dichlorofluorescein (DCF) which can specifically reveal intracellular reactive oxygen species. The protocol described here has been used in human and teleost species sperm samples. The method can be used with two approaches: (1) flow cytometry, for quantification of DCF+ cells, or (2) confocal microscopy, for the localization of ROS within the cells.

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Acknowledgments

This work was supported by the project AGL2015 68330-C2-1-R (MINECO/FEDER) and PID2019-108509RB (MICINN). Authors acknowledge Juan de la Cierva grant FJC2018-037566-I.

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Authors and Affiliations

  1. IEO, Spanish Institute of Oceanography, Planta de Cultivos El Bocal, Santander, Spain

    Vanesa Robles, Marta F. Riesco, Juan M. Martínez-Vázquez & David G. Valcarce

  2. Cell Biology Area, Department of Molecular Biology, University of León, León, Spain

    Vanesa Robles

Authors
  1. Vanesa Robles
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  2. Marta F. Riesco
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  3. Juan M. Martínez-Vázquez
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  4. David G. Valcarce
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Editors and Affiliations

  1. Ramón y Cajal Institute for Health Research (IRYCIS), Ramón y Cajal University Hospital, Centro Integrativo de Biología y Química Aplicada (CIBQA), Universidad Bernardo O’Higgins, Santiago, Chile, Madrid, Spain

    Jesús Espada

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Robles, V., Riesco, M.F., Martínez-Vázquez, J.M., Valcarce, D.G. (2021). Flow Cytometry and Confocal Microscopy for ROS Evaluation in Fish and Human Spermatozoa. In: Espada, J. (eds) Reactive Oxygen Species. Methods in Molecular Biology, vol 2202. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0896-8_8

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  • DOI: https://doi.org/10.1007/978-1-0716-0896-8_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0895-1

  • Online ISBN: 978-1-0716-0896-8

  • eBook Packages: Springer Protocols

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