Human live spermatozoa morphology assessment using digital holographic microscopy

Area of Science:

• Reproductive Biology
• Andrology
• Biomedical Engineering

Background:

• Traditional microscopy limits sperm morphology assessment to 2D.
• Digital holographic microscopy (DHM) provides a novel approach for analyzing live sperm.
• Evaluating 3D sperm parameters is crucial for accurate male fertility assessment.

Purpose of the Study:

• To assess live, intact spermatozoa morphology using DHM in fertile and infertile men.
• To introduce and evaluate novel 3D sperm morphological parameters.
• To compare sperm morphology after different sperm preparation techniques using DHM.

Main Methods:

• Digital holographic microscopy (DHM) for direct, 3D morphological analysis of spermatozoa.
• Inclusion of new 3D parameters: head height, acrosome/nucleus height, head/midpiece height.
• Comparison of sperm morphology and motility after "swim-up" and Percoll gradient centrifugation.

Main Results:

• 3D sperm parameters showed less variability in fertile men compared to infertile men.
• Significant morphological differences were observed after Percoll gradient separation and "swim-up" compared to native samples.
• DHM enabled simultaneous assessment of sperm morphology and motility.

Conclusions:

• DHM is effective for direct morphological assessment of live spermatozoa.
• Novel 3D parameters enhance the diagnostic value of semen analysis.
• DHM methodology aids in selecting morphologically sound spermatozoa and can be a valuable addition to routine sperm examination.