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

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

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...

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Bibliographic Details
Published in:Human reproduction (Oxford) Vol. 26; no. 7; pp. 1641 - 1649
Main Authors: Mallidis, C., Wistuba, J., Bleisteiner, B., Damm, O.S., Groß, P., Wübbeling, F., Fallnich, C., Burger, M., Schlatt, S.
Format: Journal Article
Language:English
Published: Oxford Oxford University Press 01-07-2011
Subjects:
Biological and medical sciences
DNA - ultrastructure
DNA Damage
Gynecology. Andrology. Obstetrics
Humans
Male
Medical sciences
Principal Component Analysis
Spectrum Analysis, Raman
Spermatozoa - ultrastructure
Raman microspectroscopy
Sperm
DNA damage
Human
Visualization
Vertebrata
Spermatozoa
Mammalia
Semen
DNA
In situ
Germinal cell
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Summary: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 which potentially could be used for nuclear DNA-based sperm selection. METHODS 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. RESULTS 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 PO4 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. CONCLUSIONS Raman microspectroscopy can provide accurate and reproducible assessment of sperm DNA structure and the sites and location of damage.
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ISSN:0268-1161
1460-2350
DOI:10.1093/humrep/der122