Complementary Tumor Diagnosis by Single Cell-Based Cytogenetics Using Multi-marker Fluorescence In Situ Hybridization...

Area of Science:

• Genetics and Molecular Biology
• Pathology
• Cytogenetics

Background:

• Multi-color fluorescence in situ hybridization (mFISH) is a key technique for diagnosing chromosomal abnormalities.
• FISH and mFISH are valuable for prenatal and pathological (tumor) diagnostics, enabling cell-by-cell analysis.
• These methods detect chromosomal gains, losses, inversions, and translocations, crucial for identifying genetic syndromes.

Purpose of the Study:

• To review the diagnostic utility and limitations of fluorescence in situ hybridization (FISH) and multi-color FISH (mFISH).
• To provide updated protocols for mFISH in formalin-fixed paraffin-embedded tissues and cytological specimens.
• To discuss data interpretation guidelines and emerging multiplexed and single-cell FISH approaches.

Main Methods:

• Review of established and novel FISH and mFISH protocols.
• Focus on applications in tumor diagnostics and single-cell analysis.
• Discussion of protocols for formalin-fixed paraffin-embedded tissues and cytological specimens.

Main Results:

• FISH and mFISH effectively detect chromosomal abnormalities in individual cells from various sample types.
• The review details protocols for preparing and hybridizing specimens, including challenging formalin-fixed samples.
• Guidelines for interpreting FISH/mFISH data and insights into advanced multiplexed techniques are provided.

Conclusions:

• FISH and mFISH are essential tools for classifying and diagnosing genetic disorders and cancers.
• Updated protocols enhance the flexibility and speed of FISH/mFISH applications.
• Future multiplexed and single-cell FISH methods promise further advancements in diagnostics.