Fig. 7

In vitro and In vivo imagining. In vitro imaging techniques were used to demonstrate the expression of YFP protein in whole FDB muscle 7 days post cDNA electroporation, and the retention of YFP post-myofiber isolation. A Whole FDB muscle expressing YFP; images were taken with a 488-nm filter cube and in phase contrast then merged to show transfection efficiency. B Whole FDB muscle YFP negative control; images were taken with a 488-nm filter cube and in phase contrast then merged. C Isolated FDB myofiber demonstrating retention of YFP. FDB myofibers were isolated and stained with mitotracker deep red and DAPI prior to imaging with a single photon confocal LSM and oil immersion ×60 objective. D Representative images of stained isolated FDB myofibers showing mitochondria in red and nuclei in blue. Two-photon excitation fluorescence microscopy and second generation harmonics were employed to generate high-resolution in vivo images of the FDB myosin structure (green) and nicotinamide-containing molecules (red) using an oil immersion ×60 objective. Representative images displayed. E Photograph of mouse being prepared for second harmonic generation microscopy of the FDB. The mouse has been tilted slightly to better show the fixing of the foot to the slide. Images were captured with the mouse in a prone position. F Myosin. G Nicotinamide. H Composite of F and G. I ×3 zoom inset of F. J ×3 zoom inset of G. K Composite of I and J