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AAT Bioquest

What are the advantages 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. 

  • Fluorescent dyes offer a narrower emission spectra and a wider variety of colors compared to chromogenic dyes, which makes multiplexing easier. 
  • Researchers can identify co-localized targets separately using fluorescent dyes for enhanced target co-localization. 
  • Fluorescent dyes make it easier to visualize both, rare and highly abundant targets, on the same slide thanks to a higher dynamic range. 
  • The substrate addition step is eliminated in fluorescent detection, resulting in fewer procedural steps. 
Additional resources

An introduction to Performing Immunofluorescence Staining

Immunohistochemistry (IHC)

Stayright™ Purple HRP Staining Kit