A Multimodal Platform for Simultaneous T-Cell Imaging, Defined Activation, and Mechanobiological Characterization.

Area of Science:

• Immunology
• Cell Biology
• Biophysics

Background:

• T-cell antigen recognition involves complex cellular rearrangements and signaling.
• Simultaneous analysis of T-cell activation processes is challenging.
• Existing methods struggle with parallel, high-quality data acquisition.

Purpose of the Study:

• To present a novel platform for multiparametric, time-resolved analysis of T-cell activation.
• To enable simultaneous quantification of T-cell receptor distribution, intracellular calcium, and T-cell-exerted forces.
• To facilitate the study of T-cell triggering using model systems.

Main Methods:

• Developed a platform integrating total internal reflection fluorescence microscopy (TIR-FM) and atomic force microscopy (AFM).
• Utilized AFM cantilevers for controlled T-cell contact with activating surfaces.
• Employed functionalized fluid-supported lipid bilayers for antigen-specific T-cell stimulation.

Main Results:

• Successfully quantified T-cell receptor (TCR) distribution via TIR-FM.
• Measured intracellular calcium dynamics.
• Recorded T-cell-exerted forces using AFM.
• Demonstrated the platform's capability with primary transgenic T-cells and cognate antigen presented by MHCII.

Conclusions:

• The developed platform enables simultaneous, high-quality, time-resolved measurements of key T-cell activation parameters.
• This integrated approach provides deeper insights into the dynamics of T-cell antigen recognition.
• The platform is suitable for studying T-cell triggering in antigen-specific model systems.