In situ visualization of damaged DNA in human sperm by Raman microspectroscopy

Mallidis C, Wistuba J, Bleisteiner B, Damm OS, Gross P, Wuebbeling F, Fallnich C, Burger M, Schlatt S (2011)


Publication Language: English

Publication Type: Journal article

Publication year: 2011

Journal

Book Volume: 26

Pages Range: 1641-9

Issue: 7

DOI: 10.1093/humrep/der122

Abstract

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.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.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)). Local spectral analysis around the DNA PO(4) backbone peak (1042 cm(-1)), showed that changes in this region were indicative of DNA damage. Wavelet decomposition confirmed both the 1042 cm(-1) shift and a second UVB susceptible region (1400-1600 cm(-1)) corresponding to protein-DNA interactions. No difference was found between observer measurements.Raman microspectroscopy can provide accurate and reproducible assessment of sperm DNA structure and the sites and location of damage.

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APA:

Mallidis, C., Wistuba, J., Bleisteiner, B., Damm, O.S., Gross, P., Wuebbeling, F.,... Schlatt, S. (2011). In situ visualization of damaged DNA in human sperm by Raman microspectroscopy. Human Reproduction, 26, 1641-9. https://dx.doi.org/10.1093/humrep/der122

MLA:

Mallidis, Con, et al. "In situ visualization of damaged DNA in human sperm by Raman microspectroscopy." Human Reproduction 26 (2011): 1641-9.

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