BACKGROUND<\/p>\n
Beyond determining the percentage of damaged sperm, current methods of DNA assessment are of limited clinical utility as they render the sample unusable. We evaluated Raman microspectroscopy, a laser-based non-invasive technique that provides detailed chemical ‘fingerprints' of cells and which potentially could be used for nuclear DNA-based sperm selection.<\/p>\n
METHODS<\/p>\n
Eight healthy donors provided ejaculates. After system optimization, a minimum of 200 air-dried sperm\/sample\/donor, prior to\/and after UVB irradiation, were assessed by two observers. Spectra were analysed by Principal Component, Spectral Angle and Wavelet Analyses.<\/p>\n
RESULTS<\/p>\n
Spectra provided a chemical map delineating each sperm head region. Principal Component Analysis showed clear separation between spectra from UV-irradiated and untreated samples whilst averaged data identified two regions of interest (1040 and 1400 cm–1<\/sup>). Local spectral analysis around the DNA PO4<\/SUB> backbone peak (1042 cm–1<\/sup>), showed that changes in this region were indicative of DNA damage. Wavelet decomposition confirmed both the 1042 cm–1<\/sup> shift and a second UVB susceptible region (1400–1600 cm–1<\/sup>) corresponding to protein–DNA interactions. No difference was found between observer measurements.<\/p>\n CONCLUSIONS<\/p>\n Raman microspectroscopy can provide accurate and reproducible assessment of sperm DNA structure and the sites and location of damage.<\/p>\n","protected":false},"excerpt":{"rendered":" BACKGROUND Beyond determining the percentage of damaged sperm, current methods of DNA assessment are of limited clinical utility as they render the sample unusable. We evaluated Raman microspectroscopy, a laser-based non-invasive technique that provides detailed chemical ‘fingerprints' of cells and … Continue reading