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

Raman microspectroscopy scanning of single live sperm is able to distinguish the human zona pellucida–bound sperm from unbound sperm.


Feng Liu, B.Sc., Yong Zhu Ph.D., Yufei Liu, Xiaobo Wang, Ping Ping Ph.D., Xinyuan Zhu, Ph.D., Hongliang Hu, Ph.D., Zheng Li M.D., Ph.D., Lin He, Ph.D.

Volume 99, Issue 3, Pages 684-689.e4, 1 March 2013



To determine if the Raman micro-spectroscopy (RMS) could be used to distinguish sperm bound to the human zona pellucida (ZP) from those unbound sperm.


Paired experiments in comparison Raman scanning feature of ZP bound and unbound sperm.


Public hospital-based clinical ART center.


Sperm samples from 10 fertile donors were used in this study.



Main outcomes measure(s):

Sperm-ZP binding, the ZP-induced acrosome reaction and scanning intensity of various regions of sperm.


The RMS found two slightly low intensity regions (800-900 and 3200-4000 cm-1) shifted to high intensity grade at the acrosome region of the ZP-bound sperm compared with unbound sperm. Moreover, principal component analysis (PCA) and statistical analysis showed that the RMS can distinguish the ZP-bound sperm from those unbound sperm.


RMS scanning of single live sperm could be used to distinguish ZP-bound sperm from those unbound sperm. Thus the RAM may be useful tool to detect normal functional sperm and to select sperm for ICSI.

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