Flow Cytometry (FACS)

Flow cytometry is a laser-based technology used to analyse the characteristics of cells or particles. It is a widely used method for analysing the expression of cell surface and intracellular molecules, characterising and defining different cell types in a heterogeneous cell population, assessing the purity of isolated subpopulations, and analysing cell size and volume. It allows simultaneous multi-parameter analysis of single cells.

Neutrophil characterisation

Neutrophils were isolated from fresh peripheral blood using commercially available isolation kit and the purity of cells analysed by FACS for neutrophil surface markers (CD45 and CD66b). Figure A.1-4 represents the gating used for characterising the neutrophils. The neutrophil population was stimulated with TNF-a for 4 hours. A difference in the LFA-1 surface antigen expression in unstimulated (B.1) and stimulated (B.2) neutrophil was observed.

THP-1 characterisation after PMA-differentiation

THP-1 (a model for human monocytes cells) on stimulation with PMA, differentiate and show macrophage-like phenotypes. The PMA-differentiated THP-1 cells were stained with macrophage surface markers (CD11; CD40) and analysed using FACS. From Figures A and B, it was observed that the expression of CD11 and CD40 was higher in PMA-differentiated THP-1 cells (red) when compared to undifferentiated THP-1 (green) cells.

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