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Related Experiment Videos

Adjusting interphase FISH results in epithelial tissue sections to whole cell complement.

M Berezuk1, J West, M Varella-Garcia

  • 1Departments of Pathology and Medicine, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Campus Box B216, Denver, Colorado 80262, USA.

Analytical and Quantitative Cytology and Histology
|May 3, 2001
PubMed
Summary

This study developed a new equation to accurately count fluorescence in situ hybridization (FISH) signals in tissue sections by correcting for differences compared to whole cell preparations.

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Area of Science:

  • Cytogenetics
  • Molecular Pathology
  • Histology

Background:

  • Accurate enumeration of fluorescence in situ hybridization (FISH) signals is crucial for diagnosing and monitoring various diseases.
  • Discrepancies often exist between FISH signal counts in whole cell preparations and tissue sections due to nuclear truncation and volumetric differences.
  • Existing methods may not fully account for these differences, potentially leading to inaccurate quantification.

Purpose of the Study:

  • To derive a mathematical equation to compensate for volumetric discrepancies between whole cell preparations and tissue sections.
  • To enable more accurate enumeration of FISH signals per cell in histologic sections.
  • To improve the reliability of FISH-based diagnostic and research applications.

Main Methods:

Related Experiment Videos

  • Calculated mean centromere signal counts in touch preparations (whole cell) and corresponding 4-6 micron paraffin-embedded tissue sections.
  • Determined mean widest nuclear diameters from the tissue sections.
  • Analyzed observed data to define volumetric relationships between tissue sections and whole cell preparations.

Main Results:

  • An equation was derived from analysis of six lung specimens.
  • The equation approximates the nuclear volume differences between whole cell preparations and tissue sections.
  • This allows for accurate quantification of the mean FISH signal count in histologic sections.

Conclusions:

  • The derived equation provides a method to correct for nuclear truncation and volumetric effects in FISH signal counting.
  • This facilitates more precise FISH signal enumeration in routine histologic sections.
  • The findings enhance the accuracy and utility of FISH in clinical and research settings.