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

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Author: Kathryn R. Perkinson, BS, HTL(ASCP)
Reviewer: Carla Shoffeitt, MSM, 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.

Continuing Education Credits

Objectives

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

Course Outline

  • Importance of Fluorescence In Situ Hybridization Testing in Cancer Care
      • Detecting Genetic Aberrations with Fluorescence In Situ Hybridization
      • Drug Therapy Based on Genetic Aberrations
      • DNA Content and Ploidy
      • Genetic Aberrations
      • Traditional Cytogenetics
      • Metaphase Fluorescence In Situ Hybridization
      • Interphase Fluorescence In Situ Hybridization in Formalin-Fixed, Paraffin-Embedded 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 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 FISH Assay
      • Equipment Required for Analysis of FISH Slides
      • Photographing FISH Slides
      • All of the following are required 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 Technique
      • Differences in Pre-Treatment
      • Differences in Digestion
      • Which of the following pre-treatment solutions are more harsh to the tissue?
      • True or False: Using the digital reading on the water bath during the FISH assay is acceptable.
      • True or False: It is important to denature the tissue and 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
      • International Standing Committee on Cytogenetic Nomenclature Reporting
      • True or False: 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: Beginner to intermediate
Intended Audience:  Clinical laboratory histotechnologists, histotechnicians, and other medical laboratory personnel interested in this subject matter. This course is also appropriate for histology and medical 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.  She holds a Bachelor of Science 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. 
The author has no conflict of interest to disclose. 
Reviewer Information: Carla J. Shoffeitt, MSM, HT(ASCP), is the Manager of the Pathology Department of Emory St. Joseph’s Hospital of Emory Healthcare in Atlanta, Georgia. She holds a Master of Science Degree in Healthcare Management and a Bachelor of Science in Biology from Troy University, Troy, Alabama. She is certified as a Histotechnician and has 25 years of experience in the field of Histology.
Course Description: This course will explore the theoretical concepts used in fluorescence in situ hybridization (FISH) testing. It will also discuss commonly used FISH methodologies, necessary equipment, and the enumeration of FISH signals. 

This course is part of:
Histology CE
Amp Probe 2
Normal ALK
Genetic translocation
Break-apart probes
DAPI