Immunohistochemistry (IHC) Basics in Histology (by LabCE)

2 P.A.C.E. contact hour(s)

(based on 164 customer ratings)

Authors: Diane L. Sterchi, MS, HTL(ASCP) and Donna M. Hedger, BS, HT(ASCP)
Reviewer: Tanisha Neely, HT(ASCP)

Course provided by LabCE.

This course introduces the various techniques that are used in the preparation and evaluation of immunohistochemistry (IHC) slides. Procedures and terminology related to IHC are also discussed and strategies for troubleshooting problems are presented.

See more courses in: Histology

Included In These Course Packages

Continuing Education Credits

P.A.C.E.® Contact Hours (acceptable for AMT, ASCP, and state recertification): 2 hour(s)
Course number 578-058-16, approved through 7/31/2018
Florida Board of Clinical Laboratory Personnel Credit Hours - Immunohistochemistry/Advanced Histologic Techniques: 2 hour(s)
Course number 20-547723, approved through 9/1/2018

Objectives

  • Explain various immunohistochemical (IHC) techniques used in the laboratory.
  • Identify appropriate quality control (QC) used while performing IHC procedures.
  • Describe slide preparation procedures used in IHC procedures.
  • Define epitope unmasking and retrieval procedures that may be used in performing IHC procedures.
  • Identify and solve problems that can occur when performing IHC procedures.

Customer Ratings

(based on 164 customer ratings)

Course Outline

  • The Role of Immunohistochemistry (IHC) in the Histology Laboratory
      • Immunohistochemistry (IHC) Uses and Considerations
      • Antibodies, Antigens, and Epitopes
  • Antibodies
      • Monoclonal Antibodies
      • Polyclonal Antibodies
      • Antibody Titering
  • Fixation
      • Tissue Handling
      • Aldehyde Fixation
      • Formalin/Aldehyde Substitutes
      • Zinc Fixatives
      • Alternative Fixatives for IHC
  • Processing and Microtomy
      • Processing Principles
      • Frozen Sections
  • Epitope Retrieval
      • General Epitope/Antigen Unmasking
      • Why Perform Epitope/Antigen Unmasking?
  • Unmasking Methods
      • Heat-Induced Epitope Retrieval (HIER)
      • Enzyme-Induced Epitope Retrieval (EIER)
  • Methods of Detection and Visualization
      • Substrates
      • Chromogens
      • 3,3’-diaminobenzidine (DAB) and 3-amino-9-ethylcarbazole (AEC)
      • Alkaline Phosphatase
      • Chromogen Enhancers
  • Immunohistochemical (IHC) Methods
      • Immunohistochemical (IHC) Techniques
      • Indirect Method
      • Direct Method
      • Avidin-Biotin Complex (ABC) Methods
      • Avidin-Biotin Complex (ABC) Method
      • Secondary Antibody
      • Peroxidase, Anti-Peroxidase (PAP) Method
  • Blocking Methods
      • General Blocking Methods
      • Avidin/Biotin Blocking
      • Serum Blocking
      • Blocking Endogenous Peroxidase Activity
      • Enzyme Blocking
  • Immunohistochemisty (IHC) Staining Reagents
      • General Reagents
      • Antibody Diluents
      • Buffers and Rinses
  • Immunohistochemical (IHC) Controls
      • Quality Control (QC)
      • Tissue Controls/Control Slides
      • Negative Quality Control (QC) Slides
      • Positive Quality Control (QC) Slides
      • System Quality Control (QC)
      • Antibody Validations
  • Troubleshooting
      • Troubleshooting
  • Record Keeping
      • Record Keeping
  • References
      • References

Additional Information

Level of instructruction: Basic to intermediate
 
Intended audience:  This program is designed as an educational and training tool for histology laboratory personnel. This course is also appropriate for students enrolled in clinical laboratory science and histology technician programs as a supplement to classroom instruction and clinical experiences.
 
Author information: Diane L. Sterchi, MS, HTL(ASCP) is a senior research associate for Covance Laboratories, Department of Discovery and Translational Services, Pathology in Greenfield, Indiana. She has worked in hospital, veterinary, and research settings in the discipline of histology. Ms. Sterchi holds an MS degree from Indiana Wesleyan University, Marion, Indiana. She has written several textbook chapters and manuals, is a frequent presenter at seminars and has conducted numerous workshops on topics including general histology, immunohistochemistry, and in situ hybridization.
 
Donna M. Hedger, BS, HT(ASCP) is a Pathology Manager at Gwinnett Hospital System in Lawrenceville, GA. She graduated from Georgia Southern University in Statesboro, Georgia with a Bachelor of Science degree in biology. She is a certified histotechnician. 
 
Reviewer information:  Tanisha N. Neely, HT(ASCP) is a freelance writer with a work history in histology, microbiology, and pharmaceutical research. Ms. Neely attended Indiana University where she earned a bachelors degree in arts & humanities from the School of General Studies and a Certificate in Histotechnology from the School of Medicine. She is a member of the National Society for Histotechnology and the Indiana Society for Histotechnology.
 
Course descripiton:  This course introduces the various techniques that are used in the preparation and evaluation of immunohistochemistry (IHC) slides. Procedures and terminology related to IHC are also discussed and strategies for troubleshooting problems are presented.

Keywords

These are the most common topics and keywords covered in Immunohistochemistry (IHC) Basics in Histology (by LabCE):

heat-induced mammal false-positives pockets ffpe controls pretreatmentantibody glutaraldehyde hazardous trap antigens hydroxymethyl b-cell one-step lymph taylor antibodybuffer antigenicity procedures silver oxygen non-staining synthetic unmasking biotin considerations methods crystals phosphatase fixative peroxide as-tr validating culture formalin phenols non-immune isothiocyanate non-self monoclonal nickel hematoxylin epitope enzymes non-specific non-formalin deparaffinization executed fuchsin technician tubules cytoplasm fails eier sodium immunohistochemical pretreatment zenkers labeled phenolic chromogen pioneer antibodies oligosaccharide heterogeneous samples anti-mouse over-fixation immunofluorescence fitc false-positive goats naphthol diazonium oven as-bi y-shaped hydrogen rinselabel autolysis buffers alcohols fluorescein resinous waterantigen unusable biotinylated cobalt 3-amino-9-ethylcarbazole paraffin-embedded levamisole amplification cells peroxidase calcium vegetable preservation avidin-biotin coverslipping trapped compounds incubation salts horseradish antibody-antigen false-negative methodology phosphates mallorys harshest blocker buffer compliance 5-bromo-4-chloro-3-indoxyl conjugated anti-peroxidase osmium b-lymphocytes xylene steamer detachment immunohistochemistry tris multi-step dilutions pre-mixed mammals substitutes ultrapure mouse wrinkles peroxidase-labeled gomoris dako proximal methanol injecting stainingblockprimary #223-galactosidase dehydrate trichrome immune polyclonals microwave glucose soluble diluents blue-black catalyzes formalin-fixed hydrolyzes ß-galactosidase solvent polyclonal suggestions immunoglobulin goat morphology enzyme antiserum enhancers aldehydes rigidity 3-diaminobenzidine competency purer epitopes sensitivity trimmed enzymatic adhesion peroxidase-based kidney chemical tumors stirring enzyme-induced fluorophore trypsin digestion bcip waterenzyme substrate bonds microtomy quench oxidase penetration methylene solutionmayers heated drawback tetroxide serum specimens immunoglobulins acetic hepatocytes stainers alcohol-based fluorescent protein fixatives vimentin conditions naphthol-as-phosphate nuclear paraformaldehyde antibodys potassium artifact isotype avidin label solvents rbcs cooker toxic inconsistency settings hier bridges antigen chromogens saline treatment concerns reagents aldehyde zinc phosphate bouins endogenous antibody alkaline serums mount alcohol pepsin metals laboratory picric rabbit mounting labels 3’-diaminobenzidine areantigen concentration azide specificity cross-links deionized
How to Subscribe
Histology CE (by LabCE)
Package of 12 online courses$65
ASCLS members save $10
Add to cart
Single online course$25
Add to cart
  • Order for instant access through any computer, any browser.
  • No shipping, faxing, or waiting for certificates.
  • Print or save your certificate of completion as soon as you've completed the course.
  • You'll have 90 days to complete your purchased courses.
Course provided by LabCE.
Image provide by Jim Burchette, HT(ASCP)


Alcohol fixed with background


IHC Counterstains


well defined IHC



 
ASCLS CE's design and platform are provided by MediaLab, Inc.