Real-time Raman microspectroscopy scanning of the single live sperm bound to human zona pellucida

摘要

Objective: To determine if Raman microspectroscopy (RMS) can distinguish sperm bound to the human zona pellucida (ZP) from those unbound sperm. Design: Paired experiments to compare Raman scanning features of ZP-bound and unbound sperm. Setting: Public hospital-based clinical assisted reproduction center. Patient(s): Sperm samples from ten fertile donors were used in this study. Intervention(s): None. Main Outcome Measure(s): Sperm-ZP binding, ZP-induced acrosome reaction, and scanning intensity of various regions of sperm. Result(s): The RMS found two slightly low-intensity regions (800-900 and 3,200-4,000 cm-1) shifted to high-intensity grade at the acrosome region of the ZP-bound sperm compared with unbound sperm. Moreover, principal component analysis and statistical analysis showed that the RMS can distinguish the ZP-bound sperm from the unbound sperm. Conclusion(s): RMS scanning of single live sperm could be used to distinguish ZP-bound sperm from unbound sperm. Thus, RMS may be a useful tool to detect normal functional sperm and to select sperm for intracytoplasmic sperm injection. ?2013 by American Society for Reproductive Medicine.