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FS FP <P2> 5 µm Figure S4. Time lapse movie showing polarized TIRF of 1:1:1 DOPC:DSPC:cholesterol supported bilayers labelled with the fluorescent probe DiI-C18. 30 um x 30um, 20 min, 20 fps/ Images were taken for 20 min at 20 fps for parallel excitation (Fs) and perpendicular excitation (Fp) of the fluorescent probe following the addition of PrP(106-126) oligomers to a final concentration of 4.1µM. Rapid switching of the polarization facilitated acquisition of near-simultaneous images for each polarization. The intensely bright dots in each panel are due to the presence of a vesicle within the pTIRF illumination field. The third panel shows the order parameter <P2> at each pixel, calculated for each frame as described in the text. Each panel represents the same 30 m x 30 m area of the bilayer, and is an expansion of the 15 m x 15 m area shown in Figure 6B.