A lung infected with γHV68 virus from a CD8KO mouse subjected to multiplexed IHC.
Interest in adopting multiplexed immunohistochemistry is quickly growing as the number of observable parameters has increased dramatically in recent years. Many systems now allow users to examine 7-100 parameters using either fluorescently tagged or metal-tagged antibodies. This technology allows users to preserve the structure of the tissue of interest and all associated cells. It also provides spatial context for cellular interactions, and allows the direct interrogation of cellular signaling. However, there are still significant experimental and computational challenges to resolve.
Computationally, there is currently a lack of suggested workflows and methodologies for the analysis of these datasets. In an attempt to ameliorate this we have intensely interrogated the current tools available in order to provide detailed methodologies for beginners and reveal important issues which may necessitate the development of novel tools. We have also worked to adapt established methods for the analysis of flow cytometry and mass cytometry data for the analysis of IHC datasets (traditional Boolean gating, dimensionality reduction algorithms, clustering algorithms, etc.).
See our Multiplexed IHC Analysis resources page for more information!