Immunophenotyping – the how, why, and where?

What is immunophenotyping?

Immunophenotyping is a test that uses fluorescently conjugated antibodies to detect the presence or absence of specific markers or antigens on the white blood cells surface, nucleus, or cytoplasm.

Certain patterns or characteristics of these antigens can assist us to classify, diagnose, and treat immunodeficiencies or types of leukaemia’s and lymphomas.

How does immunophenotyping by flow cytometry work?



Specific antibodies are added to the sample which have been tagged with fluorescent markers, which attach themselves to specific antigens on any white blood cells when the antigens are present.

These samples are then put through the cytometer to go through lasers and detectors as each cell is looked at independently.


Why is immunophenotyping used?

It can be used to test blood, bone marrow, tissue, or fluid samples to rapidly screen, quantify, diagnose disease states or processes.

The type and quantity of fluorescent antigen-antibody complexes are analysed from a multitude of cells.

The data is then input into an analysis software to be drawn up on graphs and compared against controls.

Specific classifications of leukaemia for example, would also be dependent on using the WHO classification guidelines as reference for diagnosis.



What does immunophenotyping with Applied Cytometry look like?

Using the VenturiOne software you can quickly analyse your immunophenotyping data, using a previously created workspace or playlist with all your settings preserved.

You can also use the preview plots for a quick visual overview of what is present within your data and create a workspace within a few clicks to suit your analysis needs.

Your data can be viewed in gallery or hierarchy format and using density, precedence density or colour precedence plots as shown below.

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immunophenotyping plot results table


The plots, results tables, and any custom calculations that have been created, can all be exported to a report. All of these features combined makes VenturiOne a truly flexible, eye catching and easy to use analysis platform.


Real life example 1 – Blood Monitoring for HIV

This report looks at the changes flow cytometry has brought to immunophenotyping since its increase in popularity, based in the context of the AIDS pandemic of the 1980’s.

CD4+ T lymphocyte populations and the HIV viral load are the best methods of diagnosis, and the effect of HIV and AIDS on humanity has been softened by flow cytometry development.


Real life example 2 – Leukaemia

This study looks at the importance of using immunophenotyping in acute leukaemia and finds that the lineage of the leukaemia needs to be ascertained before beginning treatment, with a risk of 1 in 4 not responding or relapsing if it is based only on the morphological diagnosis.


What’s next for immunophenotyping?

COVID-19 has given immunophenotyping a new and highly relevant place in the spotlight, with its potential to understand the differentials in response to the SARS-CoV-2 virus from different people.

It can therefore be used to create a profile (immune profiling) to adapt treatment for subsets of people who might react differently to the virus, as well as inform vaccine strategies and development in the future.











flow cytometry data interpretationfcs data analysis software immunophenotyping software analysis