The utility of sperm DNA damage assay using toluidine blue and aniline blue staining in routine semen analysis

Abstract

ObjectiveThe aim of the present study was to examine the relationship among male age, strict morphology, and sperm chromatin structure and condensation.MethodsSperm samples from a total of 100 men underwent semen analysis, and sperm chromatin structure and condensation were assessed with toluidine blue (TB) and aniline blue (AB) tests.ResultsPrevalence of strict morphology of less than 4%, and abnormal sperm chromatin structure and condensation did not show any statistically significant differences according to male age (p=0.605, p=0.235, and p=0.080). No significant correlation was demonstrated among age of male partners, strict morphology, and abnormal sperm chromatin structure using TB and AB tests. However, abnormal sperm chromatin condensation was positively associated with sperm chromatin structure (r=0.594, p=0.000) and showed negative correlation with strict morphology (r=-0.219, p=0.029).ConclusionThe tests for sperm chromatin condensation showed a significant association with strict morphology. Further study is needed to elucidate the relationship between clinical outcome and sperm chromatin tests.

Histone retention, protein carbonylation, and lipid peroxidation in spermatozoa: Possible role in recurrent pregnancy loss

“…At least 100 to 200 sperm cells were analyzed in each slide and aniline blue staining intensity was quantified as light, intermediate, and dark, thus classifying them into 3 groups of maturity. The percentage abnormal sperm chromatin condensation was calculated from the ratio of the number of stained sperm (dark + intermediate) to the total number of sperm analyzed and multiplied by 100 as described [Kim et al 2013]. …”

Optimization of microelectrophoresis to select highly negatively charged sperm

“…However, this effect was comparable in all current settings, when corrected for neutrally charged sperm. Multiple studies have reported an association between sperm chromatin compaction and abnormal sperm head morphology [61][62][63] and acrosome abnormalities [64]. Although the compaction of the nuclear chromatin and DNA integrity are two interdependent parameters [65,66], some studies have reported an association between them [67,68].…”

Use of Silver Nitrate for the Assessment of Sperm Measurements in Selected Farm and Free-Living Animal Species

“…The most frequently assessed, and the most important semen parameters include concentration, motility, and morphology of the sperm (13). These parameters are considered to be the most useful since they have been correlated with the fertility potential (14,17,18). For example, low sperm concentration, which is connected with the number of spermatozoa present in one millimetre of semen, indicates male infertility (11).…”

“…Nevertheless, its interpretations still require discussion. This is because standard staining techniques applied for the identification of semen quality, and assessment of spermatozoa are in the majority of cases limited to detection of morphological abnormality only (17,18,24), while sperm chromatin structure is of a crucial importance for fertilisation and embryo development. Routine semen assessments are the first step in the evaluation of male infertility factors, but this analysis is not considered to be sufficient for determining infertility in in vivo and in vitro conditions (17).…”