Rapid and label-free identification of normal spermatozoa based on image analysis and micro-Raman spectroscopy

Authors

  • Zufang Huang,

    1. Key Laboratory of Opto-Electronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, China
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  • Guannan Chen,

    1. Key Laboratory of Opto-Electronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, China
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  • Xiwen Chen,

    1. Key Laboratory of Opto-Electronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, China
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  • Jing Wang,

    1. Key Laboratory of Opto-Electronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, China
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  • Jinhua Chen,

    1. Fujian Provincial Hospital, Fuzhou 350001, China
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  • Peng Lu,

    1. Key Laboratory of Opto-Electronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, China
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  • Rong Chen

    Corresponding author
    1. Key Laboratory of Opto-Electronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, China
    • Phone: +86 591 83489919, Fax: +86 591 83465373===

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Abstract

Semen analysis is performed for evaluation of fertility disorders, however it is susceptible to subjectivity of investigators, and lacking of objective criterion for sperm cell quality remains a problem. There is an ongoing debate on which criteria should be employed to define normal spermatozoa. Here, the aim of our study is to evaluate the possibility of label-free and rapid identification of normal sperm cell through the use of image analysis combined with micro-Raman spectroscopy. By using a smooth-surfaced and hydrophobic substrate, spermatozoa smear was rapidly prepared for microscopic imaging and acrosome area can be clearly visualized without any extra stains; then a self-written image analysis program was utilized to segment sperm head and acrosome area and automatically calculates morphological indices. Most important, intensity ratio of 1055 cm–1 to 1095 cm–1 from the obtained Raman spectra is found to indicate a potential biomarker for assessing the sperm DNA integrity. Our preliminary study demonstrates that micro-Raman spectroscopy combined with image analysis can be a potentially useful tool for rapid and label-free identification of normal sperm cell by providing both morphological and biochemical information. (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

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