A Novel Method for Detection of Phosphorylation in Single Cells by Surface Enhanced Raman Scattering (SERS) using Composite Organic-Inorganic Nanoparticles (COINs)
Figure 5
Detection of two intracellular phosphorylation events using two different COINs simultaneously.
a) Raman spectra of the U937 cells treated with IFNγ and IL-4 simultaneously. The cells were stained with αpStat1-BFU and αpStat6-AOH simultaneously and separately. Cells were also stained with non-conjugated AOH and BFU COINs. The spectra are representative for five independent experiments. Cells were also stained with BFU and AOH that were not conjugated to antibodies. b) Extrapolated COIN spectra for treated (IFNγ+IL-4) and untreated cells (Non stim) stained with pStat1-BFU and BFU. c) Extrapolated COIN spectra for treated and untreated cells stained with pStat6-AOH and AOH. d) The Raman intensity of the Raman spectra for pStat1-BFU and pStat6-AOH COINs were calculated using the “MultiPlex” program (© Intel Corporation). The results are presented as histograms for single and double stain procedures. e) The fold change is the identified intensity of the spectra of the αpStat1 and αpStat6 conjugated COINs from treated and non-treated cells normalized to non-conjugated BFU and AOH COINs that were not conjugated to antibody. The results are the average of five independent experiments.