title: Oxidative DNA damage in human sperm can be detected by Raman microspectroscopy.
abstract: OBJECTIVE
  To determine whether Raman microspectroscopy can identify different levels of oxidative sperm nDNA damage and to corroborate the findings using an established method and an alternative but complementary spectroscopic technique.
  
  
  METHODS
  Three-way comparison of Raman profiles, Fourier transform infrared spectroscopy (FTIR) spectra, and flow-cytometric assessments of sperm nDNA damage.
  
  
  METHODS
  University-based research laboratory.
  
  
  METHODS
  Thirty-eight men attending the infertility clinic at the Centre of Reproductive Medicine and Andrology.
  
  
  METHODS
  Induction of oxidative damage by Fenton's reaction on semen samples.
  
  
  METHODS
  Raman profiles, FTIR spectra, and flow-cytometric analysis of DNA fragmentation.
  
  
  RESULTS
  Raman and FTIR spectra contained distinctive differences between untreated and fragmented nDNA sperm that were indicative of oxidative attack. The changes in Raman profiles were similar to those previously seen and corresponded to the DNA backbone. The peak attributions were corroborated by the FTIR spectra. Principal component analysis of the entire Raman spectra distinguished samples with varying degrees of damage. After determination of a cutoff value (0.63), estimation of the percentage of sperm with nDNA damage using the intensity ratio of Raman peaks (1,050/1,095 cm(-1)) correlated linearly to the flow-cytometric assessment.
  
  
  CONCLUSIONS
  Raman microspectroscopy still requires further validation but may potentially provide a means of assessing the nDNA status of a living sperm.
authors: Centre for Reproductive Medicine and Andrology, University Clinics Muenster, Germany.
authors: Victoria
authors: V
authors: Sánchez
authors: Klaus
authors: K
authors: Redmann
authors: Joachim
authors: J
authors: Wistuba
authors: Frank
authors: F
authors: Wübbeling
authors: Martin
authors: M
authors: Burger
authors: Harriette
authors: H
authors: Oldenhof
authors: Willem F
authors: WF
authors: Wolkers
authors: Sabine
authors: S
authors: Kliesch
authors: Stefan
authors: S
authors: Schlatt
authors: Con
authors: C
authors: Mallidis
mesh_terms: D015415:Biomarkers / Q000032:analysis; D004247:DNA / Q000032:analysis*; D004249:DNA Damage; D005434:Flow Cytometry; D006801:Humans; D006861:Hydrogen Peroxide; D007248:Infertility, Male / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D007501:Iron; D016014:Linear Models; D008297:Male; D015591:Microspectrophotometry; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011237:Predictive Value of Tests; D025341:Principal Component Analysis; D015203:Reproducibility of Results; D055101:Semen Analysis / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared; D013059:Spectrum Analysis, Raman; D013094:Spermatozoa / Q000737:chemistry* / Q000473:pathology
chemical_list: D015415:Biomarkers; C045076:Fenton's reagent; D004247:DNA; D006861:Hydrogen Peroxide; D007501:Iron