Fluorescence in situ hybridization (FISH) protocol in human sperm

Area of Science:

• Human genetics
• Reproductive biology
• Cytogenetics

Background:

• Aneuploidies, frequent human chromosomal abnormalities, often stem from parental meiotic errors.
• Male meiotic errors can produce spermatozoa with numerical chromosome abnormalities, increasing offspring anomaly transmission risk.
• Sperm fluorescence in situ hybridization (FISH) is a key diagnostic tool for assessing these risks.

Purpose of the Study:

• To provide a detailed, step-by-step protocol for performing sperm FISH analysis.
• To guide clinicians in processing semen samples for accurate FISH preparations.
• To enhance the understanding and application of sperm FISH in clinical reproductive genetics.

Main Methods:

• Detailed description of human semen sample processing and fixation.
• Step-by-step chromatin decondensation and denaturation procedures for sperm.
• Methodology for obtaining sperm FISH preparations and microscopic visualization.

Main Results:

• The video-article demonstrates the practical application of sperm FISH.
• It highlights critical steps for optimizing sperm FISH slide preparation and analysis.
• Focuses on the analysis of chromosomes X, Y, 13, 18, and 21 in human spermatozoa.

Conclusions:

• Sperm FISH is an established protocol for estimating numerical chromosome abnormalities in sperm.
• Accurate sample preparation and analysis are vital for reliable clinical diagnostic results.
• This protocol supports genetic reproductive counseling by assessing aneuploidy risks.