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Double Fluorescence in situ Hybridization in Fresh Brain Sections
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Rapid micro fluorescence in situ hybridization in tissue sections.

D Huber1, G V Kaigala1

  • 1IBM Research - Zürich, Säumerstrasse 4, 8803 Rüschlikon, Switzerland.

Biomicrofluidics
|June 12, 2018
PubMed
Summary

This study introduces micro fluorescence in situ hybridization (μFISH) for rapid biomarker detection in FFPE tissues. This method significantly reduces probe usage and hybridization time for faster diagnostic results.

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Area of Science:

  • Biomedical Engineering
  • Molecular Pathology
  • Genetics

Background:

  • Formalin-fixed paraffin-embedded (FFPE) tissues are crucial for diagnostics.
  • Traditional fluorescence in situ hybridization (FISH) is time-consuming and probe-intensive.
  • Need for rapid, efficient biomarker detection in FFPE samples.

Purpose of the Study:

  • To develop and validate a micro fluorescence in situ hybridization (μFISH) method for rapid cytogenetic biomarker detection.
  • To assess the efficiency of μFISH for human epidermal growth factor 2 (HER2) detection in breast FFPE tissue sections.
  • To reduce probe consumption and hybridization time in FISH assays.

Main Methods:

  • Utilized a non-contact microfluidic scanning probe (MFP) for localized FISH probe delivery on FFPE tissue sections.
  • Employed oligonucleotide FISH probes in an ethylene carbonate-based buffer for rapid hybridization.
  • Demonstrated probe recycling using hierarchical hydrodynamic flow confinements for sequential testing.

Main Results:

  • Achieved rapid hybridization: <1 min for chromosome enumeration and 10-15 min for HER2 status assessment in FFPE sections.
  • Reduced FISH probe consumption by approximately 100-fold.
  • Decreased hybridization time by 4-fold, leading to an overall assay turnaround time of <3 hours.
  • Demonstrated compatibility with standard FISH protocols and the Instant Quality FISH assay.

Conclusions:

  • Micro fluorescence in situ hybridization (μFISH) offers a rapid and efficient method for biomarker detection in FFPE tissues.
  • The μFISH method significantly reduces assay time and resource consumption.
  • μFISH holds potential for integration into pathology workflows for enhanced diagnostic and prognostic analyses.