Semen Sexing - Anatomy & Physiology

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Principle

  • X and Y chromosomes contain different quantities of DNA.
  • X-bearing sperm contain 2.8-4.2% more DNA (species dependent) than Y-bearing sperm.
  • This allows them to be sorted into two subpopulations.
  • Separation requires the uptake of DNA stain/dye called Fluorochrome into living and dead sperm.
  • X-bearing sperm take up more dye than Y-bearing sperm.
  • Vital dyes emit light at a specific wavelength when excited or activated.
    • X-bearing sperm will emit a larger signal than Y-bearing sperm.
  • This allows the sperm to be sorted by flow cytometry.

Flow Cytometry

  • Ejaculated spermatozoa are treated with a DNA dye (fluorochrome).
  • X-bearing sperm absorb more dye than Y-bearing sperm.
  • X-bearing sperm thus emit more intense light when excited by a laser.
  • Sperm are also treated with a dye that greatly suppresses the signal from dead sperm.
    • Dead sperm are therefore identified and rejected.
  • Once spermatozoa enter the flow cytometer chamber