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Mitochondria activity in frozen–thawed sperm using rhodamine 123 (R123) and propidium iodide (PI) dual staining; (a) dead sperm stained with PI; (b) live sperm with R123-stained mitochondria; (c) merged channels showing both PI and R123 stained sperm; (d) percentage of sperm with active mitochondria. Bars with the letters “a” or “b” are values with a statistically significant difference (p < 0.05, n = 4).
