A test for the practical evaluation of male fertility by acridine orange (AO) fluorescence

A new and practical test for evaluating the fertility of a male subject has been developed. Twenty-eight donors whose semen had induced at least one pregnancy resulting in a normal delivery and 61 patients attending our infertility clinic were studied. Semen smears stained with acridine orange were read on a fluorescence microscope; sperm heads appeared either green (fertile) or red ( nonfertile ). We introduce the concept of an "effective sperm count" which is obtained by multiplying the percentage of green-fluorescing sperm by the actual sperm count. Of our fertile subjects, 27 of 28 (96.4%) exhibited an effective sperm count of greater than or equal to 50 million/ml, while 60 of 61 (98.3%) infertile patients fell below this value. The percent green correlates with neither actual sperm count nor motility, indicating that this test measures a new parameter of male fertility.