Label-free detection of ConA-induced T-lymphocyte activation at single-cell level by microfluidics.
Yameng LiuXiaohu WangYuxia LanPublished in: Electrophoresis (2024)
Lymphocyte activation is critical in regulating immune responses. The resulting T-cell proliferation has been implicated in the pathogenesis of a variety of autoimmune diseases, such as SLE and rheumatoid arthritis. ConA (concanavalin A)-induced activation has been widely used in the T lymphocytes model of immune-mediated liver injury, autoimmune hepatitis, and so on. In those works, it usually requires fluorescent labeling or cell staining to confirm whether the cells are transformed successfully after medicine treatment to figure out efficacy/pharmacology. The detection preparation steps are time-consuming and have limitations for further proteomic/genomic identifications. Here, a label-free microfluidic method is established to detect lymphocyte activation degree. The lymphocyte and ConA-activated lymphocyte were investigated by a microfluidic device. According to where single cells in the sample were captured in the designed channel, lymphocyte and ConA-activated samples are differentiated and characterized by population electric field factors, 2.08 × 10 4 and 2.21 × 10 4 V/m, respectively. Furthermore, salidroside, a herbal medicine that was documented to promote the transformation, was used to treat lymphocyte cells, and the treated cell population is detected to be 2.67 × 10 4 V/m. The characterization indicates an increasing trend with the activation degree. The result maintains a high consistency with traditional staining methods with transformed cells of 15.8%, 28.8%, and 48.3% in each cell population. Dielectrophoresis is promising to work as a tool for detecting lymphocyte transformation and medical efficacy detection.
Keyphrases
- label free
- single cell
- induced apoptosis
- peripheral blood
- cell cycle arrest
- liver injury
- drug induced
- rheumatoid arthritis
- rna seq
- cell proliferation
- immune response
- healthcare
- cell death
- cell therapy
- signaling pathway
- copy number
- endothelial cells
- inflammatory response
- dendritic cells
- high glucose
- mesenchymal stem cells
- loop mediated isothermal amplification
- cell cycle
- single molecule
- flow cytometry