I have sperm cells extracted from epididymis of a mouse. I would like to check and quantify their purity.
My first idea is to use an intercalating dye like Hoechst and then see what fraction of suspension is N and what is 2N using flow cytometry. Are there any protocols for checking purity of sperm cells with flow cytometry?
Hello Ivan Pokrovac,
You can check the "purity" of your sperm suspension by several techniques.
Spermogram, to evaluate:
• The pH
• The viscosity of the semen
• The amount of sperm (count) per millilitre
• Sperm vitality: This is measured by the sperm's ability to reject the dye eosin. If the sperm is alive, it will reject it. If the sperm is dead, the pinkish colouring of eosin will persist.
• Motility
• The presence of "round cells" (also checkable by flow cytometry with DNA content, if needed)
A fluorescent intercalating agent (like Propidium iodide) to check the "purity" of your sperm cells with flow cytometry. PI+ were dead and PI- alive.
Hoping that this comment will be useful. Feel free to discuss or share some informations about your sperm suspension studies.
Ludovic Dumont, Ph.D.
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