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

Haruo Katayose, Kaoru Yanagida, Shinako Hashimoto, Hidekazu Yamada, Akira Sato

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)670-676
Number of pages7
JournalFertility and sterility
Issue number3 SUPPL. 1
Publication statusPublished - 2003 Mar 1
Externally publishedYes


  • Acridine orange
  • Diamide
  • Fertilization
  • Human sperm
  • Protamine

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynaecology


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