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Fluorescence in Situ Hybridization (FISH)-Analysis Objectives and Resources

This presentation discusses the characteristics of benign, equivocal, and malignant cell populations by FISH.  These characteristics include the pattern of DAPI staining, nuclear features, and chromosomal signals.  Appropriate filters on the fluorescent microscope are explained, as well as methods used in screening to detect malignant cells.  Common artifacts and normal appearances of cells and nondiagnostic debris as well as characteristic appearances of tumor cells are covered.  Types of chromosomal abnormalities and how they appear on FISH are also explained.  Caveats such as split signals, cell overlap, and background staining are also discussed.  (65 slides)

Objectives:

  • Describe use of FISH in the detection of neoplasia
  • Define the following:

Amplification

Aneusomy

Chromosome enumeration probes

Disomy

Filters

Fluorophore

Hemi- & homozygous deletion

Hybridization

In situ

Locus specific indicator probes

Ploidy

Polysomy

Tetrasomy

Trisomy

  • Characterize the stepwise progression through the FISH preparation process, detailing what is happening at the molecular level for each step
  • Identify characteristics of a benign, equivocal or malignant cell population by FISH (to include DAPI pattern, nuclear features & chromosomal signal pattern)

Readings:

Halling, K.C. & Wendel, A.J. In situ hybridization:  Principles and Applications.  In P.T. Cagle & T.C. Allen (Eds). Basic Concepts of Molecular Pathology.  New York: Springer, 2009.  pp. 109-118

Leonard, D.G.B. Diagnostic Molecular Pathology. Philadelphia: Saunders, 2003. pp. 46-8.

Roulston, Molecular Diagnosis of Cancer – Methods and Protocols.  Totowa, N.J. : Humana Press, 2004. pp. 77-85