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Updated: Mar 17, 2026
Qualitative and Quantitative Analysis of the Immune Synapse in the Human System Using Imaging Flow Cytometry
Published on: January 7, 2019
Multiparametric Characterization of Human T-Cell Immune Synapses by InFlow Microscopy
Guido H Wabnitz1, Yvonne Samstag2
1Institute of Immunology/Section Molecular Immunology, University of Heidelberg, Im Neuenheimer Feld 305, D-69120, Heidelberg, Germany. guido.wabnitz@immu.uni-heidelberg.de.
Insights
Analyzing immune cell communication is crucial for health. A new InFlow microscopy method efficiently quantifies protein distribution in immune synapses, improving T-cell activation studies.
Area of Science:
- Immunology and Cell Biology
- Microscopy and Imaging Techniques
Background:
- Immune cells communicate via direct contact, forming an immune synapse crucial for T-cell activation.
- Supramolecular activation clusters within the immune synapse regulate T-cell responses.
- Malfunctions in immune synapse formation negatively impact individual health.
Purpose of the Study:
- To develop a more efficient method for analyzing immune synapse formation and protein distribution.
- To overcome the limitations of traditional microscopy in studying immune synapses.
Main Methods:
- Established an InFlow microscopy approach, a hybrid of fluorescence microscopy and flow cytometry.
- Applied the method to analyze immune synapses between primary human T-cells and antigen-presenting cells.
Main Results:
- InFlow microscopy enables quantification of protein distribution in thousands of immune synapses per sample.
- The method allows for statistically powerful comparisons between different experimental conditions.
Conclusions:
- InFlow microscopy offers a high-throughput, statistically robust alternative to classical methods for immune synapse analysis.
- This technique can significantly advance research into T-cell activation and immune-related diseases.
Abstract:
Immune cells need to communicate with each other via direct cell contact formation. The contact zone has similar functions as a neuronal synapse and is therefore named immune synapse. Supramolecular activation clusters consisting of a variety of surface receptors and cytoplasmic proteins are formed within the immune synapse, which are pivotal for T-cell activation. Thus, a malfunction of immune synapse formation has detrimental effects on the healthiness of the individual.Classical confocal microscopy to analyze the supramolecular cluster formation and maturation of the immune synapse between primary human T-cells and antigen-presenting cells is time consuming and the number of cells that can be analyzed is limited. Therefore, we have established an InFlow microscopy approach for the analysis of immune synapses. InFlow microscopy is a hybrid method combining fluorescence microscopy and flow cytometry. Our InFlow microscopy method allows quantifying protein distribution in immune synapses of several hundred or even thousand cell couples in one sample. Importantly, comparisons of different samples with a strong statistical power are possible with InFlow microcopy.
