A Novel Method for Detection of Phosphorylation in Single Cells by Surface Enhanced Raman Scattering (SERS) using Composite Organic-Inorganic Nanoparticles (COINs)
Figure 4
Detection of intracellular phosphorylation signaling using COINs.
a) Flow analysis of pStat1 and Stat6 phosphorylation following treatment of U937 cells with IFNγ or IL-4 cytokines, compared to non treated (Non stim). b) Raman spectral shift intensity of BFU COIN detecting intracellular pStat1 in IFNγ and c) pStat6 in IL-4, treated and non-treated (Non stim) U937cells. Treated (Stim control) and non treated cells (Non stim control) were stained with non-conjugated BFU COIN. The spectra are representative for five independent experiments. d) Quantitation of change in Raman peak height after IFNγ and e) IL-4 treated cells compared to non-treated cells. The αpStat1 and αStat6 conjugated COINs specifically detected pStat1 and pStat6 respectively on IFNγ and IL-4 treated U937 cells compared to non-treated cells (**p<0.01). e) Fold change ratio of pStat1 and pStat6 phosphorylation in U937 treated cells stained with both BFU and AOH COINs. The changes are the average of five independent experiments.