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Related Concept Videos

Spermatogenesis01:41

Spermatogenesis

Spermatogenesis is the process by which haploid sperm cells are produced in the male testes. It starts with stem cells located close to the outer rim of seminiferous tubules. These spermatogonial stem cells divide asymmetrically to give rise to additional stem cells (meaning that these structures “self-renew”), as well as sperm progenitors, called spermatocytes. Importantly, this method of asymmetric mitotic division maintains a population of spermatogonial stem cells in the male reproductive...
Fertilization01:38

Fertilization

During fertilization, an egg and sperm cell fuse to create a new diploid structure. In humans, the process occurs once the egg has been released from the ovary, and travels into the fallopian tubes. The process requires several key steps: 1) sperm present in the genital tract must locate the egg; 2) once there, sperm need to release enzymes to help them burrow through the protective zona pellucida of the egg; and 3) the membranes of a single sperm cell and egg must fuse, with the sperm...

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Related Experiment Video

Updated: Jul 26, 2026

Analysis of Oxidative Stress in Zebrafish Embryos
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Methods to Determine Oxidative Stress in Spermatozoa.

Jacinta H Martin1,2, Leah Calvert3,4, Amanda L Anderson3,4

  • 1School of Environmental and Life Sciences, The University of Newcastle, Callaghan, NSW, Australia. jacinta.martin@newcastle.edu.au.

Methods in Molecular Biology (Clifton, N.J.)
|July 2, 2025
PubMed
Summary

Assessing oxidative stress in sperm using Dihydroethidium and Mitosox Red assays offers insights into male reproductive health. These flow cytometry methods detect reactive oxygen species, aiding fertility research and treatments.

Keywords:
Cytosolic reactive oxygen speciesDihydroethidium (DHE)Flow cytometryHumanMitochondrial reactive oxygen speciesMitosox Red (MSR)Mouse/miceOxidative stressReactive oxygen species (ROS)Spermatozoa/spermSytox® Green (SyG)

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Area of Science:

  • Reproductive Biology
  • Cellular Physiology
  • Biochemistry

Background:

  • Elevated oxidative stress in spermatozoa compromises sperm function and is linked to male reproductive pathologies.
  • Spermatozoa oxidative stress assessment offers insights into cellular and individual systemic health.

Purpose of the Study:

  • To describe two flow cytometric assays, Dihydroethidium and Mitosox Red, for detecting reactive oxygen species in spermatozoa.
  • To highlight the utility of these assays in reproductive health research and therapeutic development.

Main Methods:

  • Utilized flow cytometry to detect reactive oxygen species (ROS).
  • Employed Dihydroethidium assay for cytosolic ROS detection.
  • Employed Mitosox Red assay for mitochondrial ROS detection in both mouse and human spermatozoa.

Main Results:

  • Successfully demonstrated the capacity of Dihydroethidium and Mitosox Red assays to quantify ROS in spermatozoa.
  • Validated assays for application in both mouse and human sperm samples.

Conclusions:

  • Dihydroethidium and Mitosox Red assays are valuable tools for studying male germline health.
  • These assays support research on reproductive aging, toxicants, environmental exposures, and fertility treatments, including assisted reproduction technologies.