Time, Money, and Tissue
Simultaneous multiplex IHC can save laboratories all three
Jason Ramos | | Longer Read
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.
Read the full article now
Log in or register to read this article in full and gain access to The Pathologist’s entire content archive. It’s FREE and always will be!
Or register now - it’s free and always will be!
You will benefit from:
- Unlimited access to ALL articles
- News, interviews & opinions from leading industry experts
- Receive print (and PDF) copies of The Pathologist magazine
Or Login via Social Media
By clicking on any of the above social media links, you are agreeing to our Privacy Notice.