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Updated: Sep 27, 2026
Fluorimetric Techniques for the Assessment of Sperm Membranes
Published on: November 28, 2018
Use of diamide-acridine orange fluorescence staining to detect aberrant protamination of human-ejaculated sperm
Haruo Katayose1, Kaoru Yanagida, Shinako Hashimoto
1Department of Obstetrics and Gynecology, School of Medicine, Fukushima Medical University, Fukushima, Japan. katayose@fmu.ac.jp
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.
