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Investigation of spermatozoa and seminal plasma by Fourier transform infrared spectroscopy

Fourier transform infrared (FT-IR) spectra of human spermatozoa and seminal plasma were recorded and analyzed. Established procedure for sample preparation enabled reproducible spectra acquisition and the parameter for the control of the reproducibility of the spectra was defined. The assignment of bands was carried out using empirical approach and the origin of sperm specific "doublet", the bands at 968 cm^-1 and 981 cm^-1 was determined. Using principal component regression (PCR) model the specific regions of spectra that correlate to spermatozoa characteristics like concentration, straight line velocity (VSL) and beat cross frequency (BCF), were defined. For spermatozoa concentration amide I band, between 1700 cm^-1 and 1590 cm^-1, was defined as specific spectral region. The linear dependence of optical density of amide I band and the spermatozoa concentration was observed. Nucleic acids vibrational bands between 1119 cm^-1 and 943 cm^-1 were defined as specific spectral region for VSL determination. The relative amount of nucleic acids with respect to proteins showed linear dependence to spermatozoa straight line velocity. Spermatozoa BCF showed the best correlation to the bands of saturated hydrocarbons, between 2749 cm^-1 and 3678 cm^-1, which represent lipids and proteins. These results suggest that FT-IR spectroscopy can serve as an adjunct to conventional histopathology studies.

Fourier transform infrared spectroscopy; FT-IR spectroscopy; Human semen; Spermatozoa; Seminal plasma; Principal component regression; PCR

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