Technical Competence in Paraffin-Based Fluorescence In Situ Hybridization (FISH) (Online Course)

(based on 52 customer ratings)

Author: Kathryn R. Perkinson, BS, HTL(ASCP)
Reviewer: Jim Burchette, HT(ASCP)

This course will explore the theoretical concepts used in fluorescence in situ hybridization (FISH) testing. Commonly used FISH methodologies, necessary equipment, and the enumeration of FISH signals will also be discussed.

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Continuing Education Credits


  • Understand the theoretical concepts used in fluorescence in situ hybridization (FISH) testing.
  • Demonstrate an understanding of the methodologies involved in FISH testing.
  • Acquire an appreciation of the enumeration of fluorescence signals in paraffin-based FISH testing.
  • Identify the equipment necessary to perform FISH testing in a clinical laboratory.

Customer Ratings

(based on 52 customer ratings)

Course Outline

  • Importance of Fluorescence In Situ Hybridization (FISH) Testing in Cancer Care
      • Detecting Genetic Aberrations with Fluorescence In Situ Hybridization (FISH)
      • Drug Therapy Based on Genetic Aberrations
      • DNA Content and Ploidy
      • Genetic Aberrations
      • Traditional Cytogenetics
      • Metaphase Fluorescence In Situ Hybridization (FISH)
      • Interphase Fluorescence In Situ Hybridization (FISH) in Formalin-Fixed, Paraffin-Embedded (FFPE) Tissue
      • What type of fluorescence in situ hybridization (FISH) is performed on formalin-fixed, paraffin-embedded (FFPE) tissue?
  • Theoretical Knowledge Required for Fluorescence In Situ Hybridization (FISH) Testing
      • Chromosomes
      • Centromeres
      • Probes
      • Centromere Probes
      • Locus-Specific Probes
      • Break-Apart or Translocation Probes
      • Which of the following probe types are used to detect a translocation?
  • Equipment used in FISH technology
      • Equipment Required to Prepare and Store Reagents
      • Equipment Required for Preparation of Tissue
      • Equipment Required for Manual Performance of Fluorescence In Situ Hybridization (FISH) Assay
      • Equipment Required for Analysis of Fluorescence In Situ Hybridization (FISH) Slides
      • Photographing Fluorescence In Situ Hybridization (FISH) Slides
      • All of the following are REQUIRED in order to perform a fluorescence in situ hybridization (FISH) assay EXCEPT:
  • FISH methodology
      • Tissue Preparation
      • Common Methodologies
      • Slide Preparation
      • Pre-treatment Strategies
      • Standard Pre-treatment
      • Standard Protease Digestion
      • Standard Denaturation
      • Standard Probe Preparation and Hybridization
      • Standard Post-Hybridization and Completion
      • New Histologic-Based Fluorescence In Situ Hybridization (FISH) Technique
      • Differences in Pre-treatment
      • Differences in Digestion
      • Which of the following pre-treatment solutions are more harsh to the tissue?
      • It is acceptable to use the digital reading on the water bath during the FISH assay.
      • It is important to denature the tissue DNA and the probe DNA.
      • What happens during the hybridization step of a FISH assay?
  • Enumeration of FISH signals
      • Assessing Slide Quality
      • Control Slide
      • Patient Slide
      • HER2/neu Examples
      • Reporting Fluorescence In Situ Hybridization (FISH)
      • International Standing Committee on Cytogenetic Nomenclature (ISCN) Reporting
      • A final interpretation of FISH results is required in the report.
      • The following image is of a break apart probe for ALK translocation. Is the image depicting a normal or an abnormal sample?
  • References
      • References

Additional Information

Level of instruction: Beginning to intermediate
Intended audience:  Clinical laboratory histotechnologists, histotechnicians, and other medical laboratory personnel who have an interest in this subject matter. This course is also appropriate for histology and clinical laboratory science students, pathology residents, and practicing pathologists.  
Author information: Kathryn R. Perkinson, BS, HTL(ASCP) has been the manager of the Molecular Pathology Department of Duke University Health System in Durham, North Carolina since 1998.  She holds a  Bachelor of Science degree in biochemistry from North Carolina State University. Kathryn has over 25 years of experience in molecular biology in both the research and clinical healthcare fields at Duke University. The Molecular Pathology Department, under her leadership, has tripled in size over the past 15 years. 
Reviewer information: Jim Burchette, HT(ASCP) recently retired after 34 years at Duke University Medical Center, Durham, North Carolina. As an Immunopathology Analytical Specialist in Molecular Pathology, Jim's primary job functions included daily IHC quality control and managing the daily technical aspect of the Clinical Immunopathology operation. He has over 38 years of experience in surgical pathology histotechnology with 33 years of immunopathology specialty. Jim has an extensive experience validating new antibodies for research and diagnostic immunohistochemical use. To date, Jim has 80 publications (70 of which are peer-reviewed) and two book chapters, as well as many abstracts, posters, and technical acknowledgments in peer reviewed scientific articles. He has also served as a consultant for Triangle Biomedical Sciences (TBS), Dako North America, Leica Microsystems, and Polysciences. 
Course description: This course will explore the theoretical concepts used in fluorescence in situ hybridization (FISH) testing. Commonly used FISH methodologies, necessary equipment, and the enumeration of FISH signals will also be discussed. 

Synovial sarcoma showing green and red signals close together.

Normal ALK

Genetic translocation

Break-apart probes

PTEN edited