Abstract
Confocal Raman microspectroscopy is a valuable analytical tool in biological and medical research, allowing the detection of sample variations without external labels or extensive preparation. To determine whether this method can assess the effect of maleic acid on sperm, we prepared human sperm samples incubated in different concentrations of maleic acid, after which Raman spectra from the various regions of sperm cells were recorded. Following the maleic acid treatment, Raman spectra indicated significant changes. Combined with other means, we found that the structures and chemical compositions of sperm membranes were damaged, and even the sperm DNA was damaged by the incorporation of maleic acid. Thus, this technique can be used for detection and identification of maleic acid-induced changes in human sperm at a molecular level. Although this particular application of Raman microspectroscopy still requires further validation, it has potentially promise as a diagnostic tool for reproductive medicine.
Highlights
15 to 20% of couples face the fertility problem worldwide [1]
Several methods are currently available for the analysis of sperm nuclear DNA quality, such as sperm chromatin dispersion, the comet assay, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL), and sperm chromatin structure assay [12,13,14,15]
Each sample was further divided into three aliquots, which were assessed for sperm quality by Raman micro-spectroscopy, flow cytometry, and hypo-osmotic swelling (HOS) testing [33]
Summary
15 to 20% of couples face the fertility problem worldwide [1]. Even when fertilization is successful, poor-quality embryos, low implantation rates, and high miscarriage rates are still serious problems for some couples [2,3,4,5]. Male factor infertility is responsible for 40–50% of cases [6]. Several methods are currently available for the analysis of sperm nuclear DNA (nDNA) quality, such as sperm chromatin dispersion, the comet assay, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL), and sperm chromatin structure assay [12,13,14,15]. These methods face a similar problem, in that the processes required may lead to the destruction of the sperm sample. Is a reliable, non-invasive, and non-destructive analytical technique that provides precise information on the quality of a sperm while not affecting the integrity of the cell, thereby qualifying it for use in assisted reproductive technology (ART)
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