Use of diamide-acridine orange fluorescence staining to detect aberrant protamination of human-ejaculated sperm nuclei

Objective: To investigate the influence of human sperm nuclear chromatin on fertilization. Design: Prospective study. Setting: Assisted reproductive technology unit at a university teaching hospital. Patient(s): Fifty men starting an IVF-ET program. Intervention(s): Epifluorescent microscopic observation of human-ejaculated sperm nuclei stained with diamide-acridine orange. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of extracted sperm nucleoproteins. Main Outcome Measure(s): Usefulness of diamide-acridine orange in analysis of human sperm nuclear chromatin and fertilization ability. Result(s): There was no correlation between the semen parameters and the diamide-acridine orange observation. A positive correlation was observed between the fertilization rate after conventional IVF and the green-type increase ratio (percentage of green-pattern sperm after diamide-acridine orange staining/percentage of green-pattern sperm after acridine orange staining). Furthermore, it was suggested by SDS-PAGE that structural differences were noticed between the fertile men and the men with sperm immaturity diagnosed after diamide-acridine orange staining. Conclusion(s): Diamide-acridine orange staining was a more precise method for detecting chromatin abnormalities in human-ejaculated sperm and evaluating fertilization ability than acridine orange staining alone. This method can be used as a diagnostic tool to assess the fertilization ability of human-ejaculated spermatozoa before IVF procedures. (Fertil Steril((R)) 2003;79(Suppl 1):670-6. 2003 by American Society for Reproductive Medicine.)