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Diagnostics Precision medicine, Oncology, Histology

Time, Money, and Tissue

At a Glance

  • Immunohistochemistry is a routine part of the anatomic pathology workflow
  • By multiplexing targets, users can maximize their return from a single piece of tissue – often a precious resource
  • Testing multiple targets in a single round also saves the laboratory time and money
  • To fully benefit from the opportunities multiplex IHC offers, the next step is automation

In the decades since Coons and colleagues published their revolutionary work on immunofluorescence detection of antigens in frozen tissue (1), immunohistochemistry (IHC) has become routine in the anatomic pathology laboratory. Each target antigen of interest has been individually identified within histological sections of formalin-fixed, paraffin-embedded (FFPE) tumors or other types of tissue (2). Single-marker IHC takes advantage of the labeling capabilities of horseradish peroxidase and alkaline phosphatase enzymes, in combination with their respective reactant chromogens, to produce a colorimetric stain for visualization under a light microscope (2,3). Alternatively, fluorescent reporters – fluorochromes – can visualize the antibody-antigen interaction, either by conjugation to the primary antibody (direct immunofluorescence) or via attachment to a secondary antibody that detects the species-specific primary (indirect immunofluorescence).

More recently, IHC users have shifted from the single-marker approach to multiplexed marker detection. Multiplex IHC methods can visualize multiple target antigens within a single tissue sample and can be further subcategorized as sequential or simultaneous (4). Generally, if the primary antibodies used are from the same host species, sequential multiplex IHC is required; otherwise, they can be cocktailed and incubated simultaneously.

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About the Author

Jason Ramos

Vice President of Research and Development at Biocare Medical.

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