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AAT Bioquest

What are the disadvantages of fluorescent detection in immunohistochemistry (IHC)?

Posted May 2, 2024


Answer

Fluorescent detection is a technique used in immunohistochemistry to visualize multiple antigens simultaneously. It is based on the use of fluorophores that emit light when excited by a light source. The fluorophore is conjugated to the primary antibody for direct detection and to the secondary antibody for indirect detection. 

  • Weaker signal strength results in lower sensitivity indirect detection can be used to amplify enzyme-conjugated antibodies to enhance sensitivity. 
  • Fluorescent signals are vulnerable to photobleaching and may diminish on prolonged exposure to light. 
Additional resources

An introduction to Performing Immunofluorescence Staining

Immunohistochemistry (IHC)

Stayright™ Purple HRP Staining Kit