Non-destructive, high-resolution T cell characterization and subtyping via deep-ultraviolet microscopy.

Area of Science:

• Immunology and Cell Biology
• Biophotonics and Imaging Technology
• Biomedical Engineering

Background:

• T cell characterization is vital for immunology research, disease monitoring, and cell-based therapies.
• Existing methods like flow cytometry are often destructive and require fluorescent labels.
• Label-free imaging methods face challenges in throughput, specificity, and complexity.

Purpose of the Study:

• To introduce deep-ultraviolet (UV) microscopy as a novel, label-free imaging approach for T cell analysis.
• To demonstrate the capability of deep-UV microscopy for assessing T cell viability, activation state, and subtypes.
• To establish deep-UV microscopy as a high-throughput, non-destructive alternative to current T cell characterization techniques.

Main Methods:

• Utilized static deep-UV microscopy to image and analyze T cell viability and activation states.
• Employed dynamic deep-UV imaging to quantify intracellular activity for T cell subtyping.
• Validated deep-UV microscopy results against established flow cytometry measurements.

Main Results:

• Deep-UV microscopy accurately assessed T cell viability and activation, showing strong agreement with flow cytometry.
• Dynamic deep-UV imaging enabled rapid and precise subtyping of CD4+ and CD8+ T cells.
• The method successfully identified metabolic activity differences between T cell subtypes non-destructively.

Conclusions:

• Deep-UV microscopy provides a label-free, non-destructive, fast, and simple method for comprehensive T cell characterization.
• This technique offers a powerful tool for high-throughput immune cell analysis.
• Deep-UV microscopy has broad applications in immunology research, immune monitoring, and cell-based therapy development.