Concurrent use of flow cytometry and fluorescence in-situ hybridization techniques for detecting faulty meiosis in a human sperm sample

Mol Hum Reprod. 1998 Jan;4(1):61-6. doi: 10.1093/molehr/4.1.61.

Abstract

Routine semen analysis in an infertile patient revealed severe teratospermia associated with malformation of head and tail in 100% of the sperm cells. Flow cytometry and fluorescence in-situ hybridization (FISH) were shown to supplement routine semen analysis by providing information on the sperm chromatin. Using flow cytometry, propidium iodide-stained spermatozoa from the same sperm sample were compared with a normal reference pool, and with human lymphocytes. The results point to a population of diploid sperm cells rather than to mature haploid spermatozoa. Numerical chromosomal abnormalities of the spermatozoa were subsequently evaluated using FISH. A total of 1000 sperm cells were scored for X and Y chromosomes, and an additional 1128 sperm cells for chromosome 18. Aneuploidy of chromosomes X and Y was revealed in 96.9% of the cells and of chromosome 18 in 90.3% of the cells. Non-disjunction of chromosome X and Y in meiosis I and II occurred in 54.8 and 2.7% of the sperm cells respectively. Non-disjunction in both meiosis I and II occurred in 39.4% of the sperm cells. A normal haploid pattern for chromosomes X and Y was observed in only 3.1%, and for chromosome 18 in 9.7%, of the cells. Using three colour FISH for the sex chromosomes and for chromosome 18, diploidy was demonstrated in 19.4% of 500 sperm cells and aneuploidy in virtually all sperm cells (99.2%). The use of flow cytometry and FISH in cases where genetic and developmental chromatin abnormalities are suspected is a valuable adjunct to other available techniques, and can guide the clinicians to decide which samples are unsuitable for intracytoplasmic injection.

Publication types

  • Case Reports

MeSH terms

  • Adult
  • Aneuploidy
  • Chromatin
  • Chromosomes, Human, Pair 18 / genetics
  • Flow Cytometry / methods*
  • Humans
  • In Situ Hybridization, Fluorescence / methods*
  • Male
  • Meiosis*
  • Nondisjunction, Genetic
  • Oligospermia / pathology*
  • Spermatozoa / abnormalities*
  • Spermatozoa / cytology
  • Spermatozoa / physiology
  • X Chromosome / genetics
  • Y Chromosome / genetics

Substances

  • Chromatin