title: Rapid and label-free identification of normal spermatozoa based on image analysis and micro-Raman spectroscopy.
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.
authors.affiliation: 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.
authors.forename: Zufang
authors.initials: Z
authors.lastname: Huang
authors.forename: Guannan
authors.initials: G
authors.lastname: Chen
authors.forename: Xiwen
authors.initials: X
authors.lastname: Chen
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Jinhua
authors.initials: J
authors.lastname: Chen
authors.forename: Peng
authors.initials: P
authors.lastname: Lu
authors.forename: Rong
authors.initials: R
authors.lastname: Chen
mesh_terms: D006801:Humans; D007091:Image Processing, Computer-Assisted; D008297:Male; D008853:Microscopy; D013059:Spectrum Analysis, Raman; D013094:Spermatozoa / Q000166:cytology*; D013997:Time Factors
keywords: DNA integrity; acrosome; image analysis; micro‐Raman spectroscopy; sperm cell