Figure 1.
Schematic showing the bioanalytical tools required for evaluation of the polyQ structure in vivo.
Figure 2.
Experimental and calculated X-ray fibre diffraction pattern for fibres of the poly-L-glutamine peptide, D2Q15K2.
The experimental X-ray fibre diffraction pattern has the fibre axis running vertical and the strong characteristic cross-β reflection at 4.75 Å and equatorial reflections at 8.13, 4.05, Å (see labels) 3.55 and 3.25 Å. Top right hand corner shows calculated diffraction pattern from coordinates [22] and shows an excellent match in signal positions and relative intensity.
Figure 3.
Raman and predicted M-CARS spectra of fibres of D2Q15K2.
(i) Raman spectrum of the fibres of D2Q15K2 as evaluated by X-ray diffraction. (ii) Predicted M-CARS spectra (solid line corresponds to 50%, and dashed line corresponds to 20% non resonance contribution.)
Figure 4.
CARS and fluorescence microscopy images of the nematode, C. elegans (Q40-YFP) at 1 day old adult (day 4).
(i) Image of the lateral view obtained by CARS microscopy (1657 cm−1); where the CARS intensity (in arbitrary units) is shown by the scale bar and the fluorescence intensity is shown by green contours. Four regions are identified (a,b,c,d) which are shown in an expanded form in figures (ii) to (vi) (ii) Expanded view of regions a, b, c from figure (i) where the CARS signal (1657 cm−1) intensity is greatest. The axes are in microns and correspond to the axes shown in figure (i), other than for region ‘c’ which is rotated; the axes are in microns. (iii) Plot of the region identified as ‘a’ in figures (i) and (ii) against intensity (in arbitrary units). The fluorescence signal is identified in green/green contours and the CARS signal (1657 cm−1) black-red-yellow with a scale as in figure (ii). (iv) Plot of the region identified as ‘c’ in figures (i) and (ii) against intensity (in arbitrary units). Note: the rotation, the cuticle and hypodermis of the C. elegans nematode is to the right hand side of the figure, rather than at the bottom of the figure as in (ii) expanded view ‘c’.