Figure 1.

Pre-proccesing of spectra and selection of appropriate wavelengths. A) Raw fluorescence emission spectra of various AChR-free liposomes with varying molar ratios of DOPA/DOPC (ranging from 0 to 60%) and two AChR-containing liposomes with DOPA/DOPC (20:40). Two replicas from independent experiments are shown. In all samples, NBD-Chol and PyPC concentrations were kept at 40 and 5%, respectively. The final lipid concentration and the lipid to protein molar ratio in the AChR-containing liposomes were 40 μM and 500:1, respectively. Excitation wavelength = 340 nm. Temperature = 25°C. B) Pre-processed spectra obtained by smoothing throughout the Savitsky-Golay procedure (with a window width of 8 points and a second order polynomial fit) and normalizing to the highest value of each spectrum (corresponding to the first PyPC emission band at 374 nm). The wavelengths further employed for the multivariate analysis are outlined in the dotted rectangle. It is clear that the sixteen spectra in each panel are difficult to identify due to their high degree of overlap; this is precisely what multivariate analysis dissects so effectively. Where possible, spectra have been identified with the sample numbers appearing in Table 1.