Detection of the T cell activation state using nonlinear optical microscopy

Nonlinear microscopy imaging provides qualitative and quantitative distinction between activated and na ïve T‐lymphocytes based on the lipid body content of the cells, correlated with cholesterol and lipid spectra. The ability to monitor the activation state of T ‐cells during immunotherapy is of great importance. Although specific activation markers do exist, their abundance and complicated regulation cannot definitely define the activation state of the cells. Previous studies have shown that Third Harmonic Generation (THG) imaging could distinguish betwe en activated versus resting microglia and healthy versus cancerous cells, mainly based on their lipid‐body profiles. In the present study, mitogen or antigen‐stimulated T‐cells were subjected to THG imaging microscopy. Qualitative and quantitative analysis showed statistically significant incr ease of THG mean area and intensity in activated versus resting T‐cells. The connection of THG imaging to chemical information was achieved using Raman spectroscopy, which showed significant differences between the activation processes and controls, correlating of THG signal area with cholesterol and lipid compounds, but not with triglycerides. The obtained results suggested a potential employment of nonlinear microscopy in evaluating of T‐cell activation, which is expected to be largely appreciated in the clinical practice.
Source: Journal of Biophotonics - Category: Physics Authors: Tags: FULL ARTICLE Source Type: research