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Related Experiment Videos

The use of a cell-cycle phase-marker may decrease the percentage of errors when using FISH in PGD.

A Pujol1, J Benet, M Campillo

  • 1Departament de Biologia Cel.lular, Fisiologia i Immunologia, Unitat de Biologia, Facultat de Medicina, Universitat Autònoma de Barcelona, Barcelona, Spain.

Cytogenetic and Genome Research
|June 26, 2004
PubMed
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Fluorescence in situ hybridization (FISH) on embryos can lead to misdiagnosis due to DNA replication. Cell cycle markers are recommended to accurately interpret FISH signals and avoid errors in genetic analysis.

Area of Science:

  • Genetics and Genomics
  • Cell Biology
  • Reproductive Biology

Background:

  • Fluorescence in situ hybridization (FISH) is crucial for analyzing chromosome abnormalities in preimplantation embryos.
  • FISH is applied in various scenarios including carrier screening, sexing, and aneuploidy screening for specific patient groups.
  • Accurate interpretation of FISH signals is vital, especially in asynchronously dividing interphase blastomeres.

Purpose of the Study:

  • To investigate whether cell cycle stage influences the interpretation of FISH results.
  • To determine the impact of DNA replication on FISH signal characteristics.
  • To assess the potential for misdiagnosis in proliferating cells analyzed by FISH.

Main Methods:

  • Compared FISH signal characteristics using locus-specific, subtelomere, and centromere probes.

Related Experiment Videos

  • Analyzed signal patterns in non-dividing Sertoli cells (G0/G1) and proliferating lymphocytes (G1, S, G2).
  • Quantified the percentage of nuclei with aberrant signal counts in dividing versus non-dividing cells.

    • Main Results:

    • A significant difference (P < 0.05) in the percentage of nuclei with unexpected signal counts was observed between dividing and non-dividing cells.
    • Approximately 10.8% of double FISH signals in dividing cells were attributed to DNA replication.
    • The replication sequence observed was locus-specific region, followed by telomere, and then centromere regions.

    Conclusions:

    • DNA replication can lead to misinterpretation of FISH signals, potentially causing misdiagnosis in up to 7% of proliferating cells.
    • Cell cycle phase-specific markers are essential to accurately determine the cell stage during FISH analysis.
    • Implementing cell cycle markers can prevent diagnostic errors and improve the reliability of FISH-based genetic assessments.