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Human sperm often contain vacuoles, and it is therefore necessary to determine whether vacuoles in the chromosomal region are responsible for DNA deterioration. To that end, the present study developed a novel clinical examination technique, translucent staining, for the visualization of vacuoles. Highly diluted Reactive Blue 2 (RB2, 100 pmol/L) faintly stained the sperm head to reveal a translucent bluish body with toneless spots. Differential interference contrast showed the vacuoles as shaded-relief images. In the same, field of view the localization and the shape of the toneless spots were coincided with the shaded-relief images.
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