Fig. 7. Quantification of polycarbonate filter penetration assay. (A) Approximately 2.1x10⁴ Acr3-EGFP sperm were assayed for penetration through polycarbonate filters. The average number of sperm (± s.e.m.) in the lower and upper chambers was determined by FACS and hand counting. The number of sperm recovered from the upper chamber was 123.3±38.1, 141.7±46.8, 1356.7±165.9, and 2165.0±188.0, after passage through filters with pore sizes 1.2, 3, 5 and 12 µm, respectively. (B) The acrosome status of Acr3-EGFP sperm was determined before (epididymal) and after (treated) passage through the glass bead column (controls) by FACS. Following the 30 minutes filter penetration assay, the percent of acrosome-reacted sperm in the lower (blue) and upper (red) chambers was determined by FACS and confirmed morphologically.