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Updated: Sep 2, 2025
Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques
Published on: April 22, 2010
An imaging mass cytometry immunophenotyping panel for non-human primate tissues
Paula Niewold1, Marieke E Ijsselsteijn2, Frank A W Verreck3
1Department of Infectious Diseases, Leiden University Medical Centre, Leiden, Netherlands.
Insights
Developing an imaging mass cytometry panel is crucial for understanding cellular organization in non-human primate tissues. This 18-marker panel visualizes leukocyte subsets, aiding research in health and disease outcomes.
Area of Science:
- Cellular Biology
- Immunology
- Translational Research
Background:
- Cellular spatial organization is vital for function in health and disease.
- Imaging mass cytometry (IMC) preserves spatial context for analyzing cell relationships.
- Preclinical models, especially non-human primates, are essential for translational research due to physiological similarities to humans.
Purpose of the Study:
- To develop a pipeline for selecting and incorporating antibodies for imaging mass cytometry on non-human primate formalin-fixed paraffin-embedded (FFPE) tissues.
- To create a high-dimensional IMC panel for analyzing leukocyte subsets in non-human primate tissues.
- To advance the understanding of cellular organization and its impact on disease outcomes using non-human primate models.
Main Methods:
- Development of a novel antibody selection and incorporation pipeline for IMC.
- Creation of an 18-marker backbone IMC panel.
- Application of the panel to visualize leukocyte subsets in rhesus and cynomolgus macaque tissues.
Main Results:
- Successful development of a method for antibody selection for non-human primate FFPE tissue IMC.
- Establishment of an 18-marker IMC panel.
- Demonstrated visualization of diverse leukocyte subsets within non-human primate tissues.
Conclusions:
- The developed pipeline enables effective IMC panel creation for non-human primate tissues.
- The 18-marker panel facilitates detailed analysis of cellular organization in macaques.
- This tool will enhance research into disease mechanisms and outcomes in translational studies.
Abstract:
It has recently become clear that spatial organization contributes to cellular function and that expanding our knowledge on cellular organization is essential to further our understanding of processes in health and disease. Imaging mass cytometry enables high dimensional imaging of tissue while preserving spatial context and is therefore a suitable tool to unravel spatial relationships between cells. As availability of human tissue collected over the course of disease or infection is limited, preclinical models are a valuable source of such material. Non-human primate models are used for translational research as their anatomy, physiology and immune system closely resemble those of humans due to close evolutionary proximity. Tissue from non-human primate studies is often preserved large archives encompassing a range of conditions and organs. However, knowledge on antibody clones suitable for FFPE tissue of non-human primate origin is very limited. Here, we present an imaging mass cytometry panel development pipeline which enables the selection and incorporation of antibodies for imaging of non-human primate tissue. This has resulted in an 18-marker backbone panel which enables visualization of a broad range of leukocyte subsets in rhesus and cynomolgus macaque tissues. This high-dimensional imaging mass cytometry panel can be used to increase our knowledge of cellular organization within tissues and its effect on outcome of disease.
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