Journal of Medical Diagnostic Methods
Open Access

Department of Obstetrics and Gynaecology, Ichikawa General Hospital, Tokyo Dental College, 5-11-13 Sugano, Ichikawa, Chiba 272- 8513, Japan

Publications

• Research Article   
  Re-Evaluation of Sperm Chromatin Structure Assay (SCSA)
  Author(s): Satoru Kaneko* and Kiyoshi Takamatsu
  
  The Sperm Chromatin Structure Assay (SCSA) based on the polychromatic fluorescence of Acridine Orange (AO) is widely employed to observe DNA damages in human sperm nuclei. To re-evaluate the principle of the SCSA, human motile sperm with a negative DNA fragmentation rate of 87% were prepared as the Negative Control (NC) and the Positive Control (PC) was prepared through heat denaturation of the NC. The AO stainability for the controls was compared on the basis of microscopic, electrophoretic, and flow cytometric observations. Human motile sperm were separated by means of Percoll density gradient centrifugation and subsequent swim up. Then, DNA fragmentation was observed by the aid of Single-cell pulse-field gel electrophoresis. We found that the AO molecule emitted red and green fluorescence depending on the excitation wavelength. Both controls were change.. View More»
  DOI: 10.35248/2168-9784.23.12.399

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