Comparison of Special Stains for Analysing the Morphology of Sperm: A Cross-sectional Study

Abstract

Introduction: In India, male infertility constitutes 40% of infertility cases in males, which has become a serious problem in developing countries. Numerous studies and advanced techniques have been identified in assessing sperm morphology. The detection of sperm morphology is a simple and widely accepted method for determining sperm viability. It assists in selecting appropriate treatments for male infertility in assisted reproductive techniques. Infertility issues can arise in healthy males due to altered sperm morphology. Aim: To analyse the morphology of sperm using basic histopathology stains such as Haematoxylin and Eosin (H&E), Giemsa (G), Eosin-Nigrosin (EN), and Papanicolaou (Pap) stains. Materials and Methods: A cross-sectional study was conducted from April 2022 to June 2022 at the Department of Clinical Pathology, Sree Balaji Medical College and Hospital, Chennai, Tamil Nadu, India. Thirty semen samples were collected from healthy males and stained with H&E, Giemsa, Eosin-Nigrosin, and Papanicolaou stains. The results were observed and tabulated based on sperm shape, size, and motility using Kruger’s strict morphology method. A comparison was made to identify the fastest and most cost-effective method among the four stains. Descriptive statistics were used for the comparison. Results: Among the 30 semen samples, H&E and Papanicolaou stain methods were found to be rapid and cost-effective for analysing sperm morphology compared to the other stains (p-value <0.001). The findings included cases of Normozoospermia (18), Oligozoospermia (9), Necrozoospermia (1), and Teratozoospermia (2). In the Papanicolaou stained samples, the mean sperm head length (μm) was 8.92a±0.5, head width (μm) was 4.48a±0.33, head perimeter (μm) was 27.69a±1.85, head area (μm²) was 33.96a±3.74, tail length (μm) was 44.31a±2.02, sperm length (μm) was 53.24a±2.18, with no cytoplasmic droplets greater than half the size of the head and two vacuoles observed. Papanicolaou stain was the least expensive stain (9.50rs) compared to the other stains used for assessing sperm morphology. Conclusion: Papanicolaou stain is a simple and cost-effective method for analysing sperm morphology compared to other special stains. However, EN Stain is used to detect the morphology of live sperm. Sperm morphology assessment is necessary to monitor the causes of male infertility and determine appropriate treatment modalities.