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Related Experiment Videos

Sequential application of interphase-FISH and CGH to single cells.

Sabine Langer1, Jochen B Geigl, Rainer Gangnus

  • 1Institut für Humangenetik, Technische Universität München, Munich, Germany.

Laboratory Investigation; a Journal of Technical Methods and Pathology
|February 8, 2005
PubMed
Summary

This study introduces a novel method for verifying single-cell comparative genomic hybridization (CGH) results. By sequentially applying fluorescence in situ hybridization (FISH) and CGH to the same cell, researchers can now accurately analyze and confirm genomic data from individual cells.

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Area of Science:

  • Genomics
  • Cell Biology
  • Molecular Diagnostics

Background:

  • Comprehensive genomic analysis of single cells is crucial for tumor genetics, clinical diagnostics, and forensic applications.
  • Existing whole-genome amplification and comparative genomic hybridization (CGH) protocols for single cells lack verification methods due to cell destruction during analysis.

Purpose of the Study:

  • To develop and demonstrate a novel sequential approach for analyzing and verifying single-cell genomic data.
  • To enable sophisticated cell selection based on initial interphase fluorescence in situ hybridization (FISH) signals before comprehensive genomic analysis.

Main Methods:

  • Sequential application of interphase fluorescence in situ hybridization (FISH) and laser microdissection for cell selection.
  • Linker-adaptor PCR for whole-genome amplification of selected single cells.

Related Experiment Videos

  • Comparative genomic hybridization (CGH) analysis on amplified single-cell DNA.

    • Main Results:

    • Successfully demonstrated the feasibility of sequential FISH and CGH on the same single cell.
    • Enabled verification of single-cell CGH results, overcoming previous limitations.
    • Showcased application in analyzing cells with chromosomal abnormalities (trisomies 13, 18, 21), leukemia, tumor cells, and tissue sections.

    Conclusions:

    • The sequential application of FISH and CGH to the same single cell provides a robust method for controlling and verifying genomic analysis.
    • This approach enhances the reliability of single-cell genomic studies in diagnostics and research.
    • Offers new possibilities for precise cell selection and detailed genomic profiling.