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The Four “i’s” of Immunophenotyping

Posted on: June 20, 2018

Immunophenotyping is a type of flow cytometry-based analysis in which cells are stained with fluorescently-conjugated antibodies for specific surface and intracellular molecules that define different immune cell populations. Immunophenotyping has been widely used for decades since the advent of cytometers that can detect four colors, and today’s state-of-the-art cytometers allow for concurrent immunophenotypic analysis of numerous cell subsets.


Immunophenotyping has been critical to immuno-oncology research because it allows researchers to monitor changes in the immune system, including potent anti-tumor responses or unexpected off-target responses.

Consider these four “I’s” of immunophenotyping as you plan your next research project.

Immune System Profile

Immunophenotyping allows you to gauge the overall status of the immune system, from B and T cell subsets, to monocytes/macrophages and dendritic cells. Immunophenotyping assays are usually done using peripheral blood and can be optimized to use whole blood or peripheral blood mononuclear cells. 

In Depth Analysis

Immune cell subpopulations can be measured using immunophenotyping, so you can analyze different B cell or T cell subsets to assess antigen-specific effector or memory responses or determine changes in dendritic cell subsets in the periphery during a specific treatment.


Treatment with immune-modifying drugs or biologics is becoming more common in oncology and the management of other chronic conditions. These treatments alter immune responses by blocking or activating different immune cell functions but can also interact with immune cells in unexpected ways to trigger unfavorable responses. Immunophenotyping allows researchers to track changes in the immune system that cause undesirable or dangerous responses.


Interleukins are a large class of molecules produced by immune and non-immune cells to activate immune and inflammatory responses. These molecules are produced within cells and can be measured by intracellular staining as part of an immunophenotyping protocol. Monitoring specific interleukin responses is critical to gauging if immune responses are productive or destructive.

Gain insight into the immune system with immunophenotyping assays. If these assays are new to your research program, consider working with flow cytometry experts or contract research organizations to assure that you are using the appropriate samples and protocols for your immunophenotyping goals. 


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