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. 2019 Nov 20;7:295. doi: 10.3389/fcell.2019.00295

FIGURE 2.

FIGURE 2

PSA-FITC labeling for the assessment of acrosomal integrity. (A) Representative sperm PSA-FITC labeling of one donor sample. PSA-FITC labeling was conducted in purified sperm by density gradient centrifugation (DGC; upper left), sperm incubated with capacitation medium (CAP; upper right) and sperm in which the acrosome reaction was induced by incubation with a calcium ionophore A23187 (AR; lower left). Vehicle control is also shown (DMSO control; lower right). Acrosomal vesicles are shown in green and the sperm cells with reacted acrosomal vesicles are indicated by an arrowhead. Sperm nuclei are stained with DAPI (blue). Scale bar = 5 μM (B) Quantification of the number of sperm with an intact acrosome, partially intact acrosome and reacted acrosome in the three fertile donors used for this study. The number of spermatozoa was counted after PSA-FITC labeling on DGC, CAP, AR, and DMSO sperm. Significant differences in the percentage of sperm cells with reacted acrosomes are indicated with stars (p-value < 0.01; one-way ANOVA with Holm–Sidak correction).