Fitc psa l 0770

To evaluate the integrity of the acrosome, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin (FITC-PSA, L0770; Sigma-Aldrich) was used. Initially, a 30-µL smear was prepared on a glass slide for each sample. After drying, the samples were fixed at room temperature with methanol for 30 minutes. Then, 50 µL of FITC-PSA solution was poured on each slide and incubated for 30 minutes. Finally, the stained slides were washed with distilled water and were evaluated using a BX51 fluorescence microscope (Olympus; excitation at 450–490 nm, emission at 520 nm) with a magnification of ×1,000. For each slide, 200 sperm were examined. Sperm with bright green fluorescence in the acrosome area were identified as having intact acrosomes, while sperm with no green or pale green fluorescence near the equator were identified as acrosome-reacted (Figure 5) [21 (link)].