基于拉曼光谱评估人类精子存活能力的研究

Non-invasive evaluation of human sperm vitality using Raman spectroscopy

目的：探讨拉曼光谱用以鉴别正常精子和死精子的可行性。方法：以10例捐精者正常精子为实验组，5例死精子为对照组。伊红染色法鉴别死活精子，分别按照形态学对精子核仁、顶体、颈部进行拉曼光谱扫描，比较研究组和对照组拉曼光谱强度。结果：精子的拉曼光谱主要由500-650／cm、1000～1200／cm2个区域组成，峰强集中在550／cm及1100／cm左右，不同扫描位点的拉曼光谱无统计学差异（p〉0．05）。死精子在拉曼光谱强度上明显强于正常精子：正常精子强度普遍〈300IU，死精子强度〉400IU。结论：拉曼光谱可对精子的存活能力提供新的评估方法，并为进一步筛选正常精子用于辅助生殖技术开拓新思路。

Objective： To investigate if Raman spectroscopy can be used to distinguish live and dead human sperm. Methods： Five necrospermic semen samples were selected as experimental group, while 10 normal semen samples were set as control group. Eosin-Y test was firstly used to distinguish live and dead sperm, then Raman spectroscopic scanning was set on nucleus, acrosome, and neck of sperm, respectively. Results： Raman spectrum of sperm was mainly consisted of two areas at 500-650/cm and 1 000-1 200/cm, and the spectral peaks concentrated around 550/cm and 1 100/cm（P〉0.05）. The Raman spectra had a significantly higher intensity in dead sperm （〉400 IU） than in live sperm （〈300 IU）（P〈0.05）, while no obvious differences were found in nucleus, acrosome and neck. Conclusion： Raman spectroscopy provides a new evaluation method for human sperm vitality, and may serve as a new approach to select live sperm for assisted reproductive technique.