Fluorescence Microscopy Imaging of Spermatozoa (IMAGE)
Caption
Illustrations (a) and (b) show samples stained with a mix of fluorochromes (Hoechst, SYBR14, and Propidium Iodide). Illustrations (c) and (d) display samples stained with a second fluorochrome mix (Draq5, Calcein Violet, and PI). Both fluorochrome mixes are used to assess sperm viability, a crucial parameter for evaluating semen quality before and after freezing. The results obtained from both analyses were comparable.
(a) A living spermatozoon, where the nucleus is positive for Hoechst (blue), a DNA marker, and SYBR14 (green), also a marker for live cell DNA. (b) A dead spermatozoon, where the nucleus is marked blue by Hoechst and red by Propidium Iodide (PI), which stains the DNA of dead cells (permeable membrane) in orange. (c) The cell indicated by the arrow is dead, as it is also positive for PI. It is marked red by Draq5, another type of DNA marker. The second spermatozoon is alive, its DNA stained red, and its flagellum appears blue due to Calcein Violet staining, indicating cell viability. (d) A moribund spermatozoon with double staining: PI and Calcein Violet.
Credit
University of Liège / Sophie Egyptien
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