Comparison of Diff-Quick and Spermac Staining Methods for Sperm  Morphology Evaluation

Lincoln Bastos  Farias Junior; André Rodrigues  da Cunha Barreto-Vianna; Mariana Duque  de Mello; Alexandre Leseur  dos Santos; Cristiane da  Fonte Ramos; Paula  Fontoura

doi:10.18502/jri.v24i3.13272

Lincoln Bastos Farias Junior Rio de Janeiro Sperm Bank (Banco de Sêmen do Rio de Janeiro), Rio de Janeiro, Brazil
André Rodrigues da Cunha Barreto-Vianna Federal University of Paraná-Palotina, Paraná, Brazil
Mariana Duque de Mello Rio de Janeiro Sperm Bank (Banco de Sêmen do Rio de Janeiro), Rio de Janeiro, Brazil
Alexandre Leseur dos Santos Federal University of Paraná-Palotina, Paraná, Brazil
Cristiane da Fonte Ramos State University of Rio de Janeiro, Rio de Janeiro, Brazil
Paula Fontoura Rio de Janeiro Sperm Bank (Banco de Sêmen do Rio de Janeiro), Rio de Janeiro, Brazil

DOI: https://doi.org/10.18502/jri.v24i3.13272

Keywords: Human, Morphology, Spermatozoa, Staining.

Abstract

Background: The objective of the current study was comparing the impact of two staining techniques on semen morphological parameters and their influence on patient diagnosis. The ideal staining method should preserve cell integrity while providing detailed information.

Methods: Semen samples from fifty men were stained using Diff-Quick or Spermac methods. Morphological parameters were classified based on the Tygerberg criteria, and final diagnosis was according to WHO manual guidelines. Statistical analysis was performed through conducting paired t-tests or Wilcoxon rank-sum tests, with GLIMMIX and Fisher's exact test for determining the significance (p≤0.05).

Results: Both staining methods highlighted head and tail regions, with Spermac offering better visualization of the midpiece. Spermac demonstrated fewer normal spermatozoa (2.8±0.3%) compared to Diff-Quick (3.98±0.4%; p=0.0385). Midpiece abnormalities were more evident with Spermac (55.7±2.1%) than Diff-Quick (24.8±2.0%; p<0.0001). No significant difference was found in head and tail abnormalities (p>0.05).

Conclusion: Diff-Quick staining resulted in a higher proportion of normal spermatozoa, primarily due to its midpiece evaluation. The choice of staining method significantly impacts the diagnosis of infertile males. These findings have important implications for clinical practice and future research, suggesting the need for further investigations to assess different staining methods and determine optimal diagnostic thresholds.