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Real-time Raman microspectroscopy scanning of the single live sperm bound to human zona pellucida
Feng Liu1, Yong Zhu, Yufei Liu
1Department of Urology, Renji Hospital, Shanghai Human Sperm Bank, Sperm Development and Genetics Laboratory, Shanghai Institute of Andrology, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
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.
