A Novel Method for Detection of Phosphorylation in Single Cells by Surface Enhanced Raman Scattering (SERS) using Composite Organic-Inorganic Nanoparticles (COINs)
Figure 2
a) Image of the “Integrated Raman BioAnalyzer” - IRBA. The arrow indicates the placement of the chamber with the sample prior to insertion into the apparatus. b) Generic configuration of Raman microscopic setup. c) Optimization of scans using IRBA. Wells containing COINs were scanned with a laser beam of 1 µm using a matrices of 5×5, 10×10, 15×15, 17×17 and 20×20 at 100 µm distances. The spectra are indicated (left) and the calculated peak heights are represented as histograms (right). The experiments were performed 3 times in duplicates. The peak heights for the 15×15 and 17×17 are significantly different from the 5×5, 10×10 and 20×20; **p<0.01. d) Raman intensity of spectra from cells stained with different concentrations of αCD54-AOH-COIN (red - 0.5 mM, blue – 0.25 mM and yellow – 0.1 mM) and AOH-COIN (purple - 0.5 mM, green – 0.25 mM and orange – 0.1 mM), scanned by IRBA (left). Quantitation of the Raman peak height from the spectra observed illustrated as histograms *p<0.05 and **<0.01. The experiment was performed three times in duplicates.