Determination of Myosin Filament Orientations in Electron Micrographs of Muscle Cross Sections

An automated image analysis system for determining myosin filament azimuthal rotations, or orientations, in electron micrographs of muscle cross sections is described. The micrographs of thin sections intersect the myosin filaments which lie on a triangular lattice. The myosin filament profiles are variable and noisy, and the images exhibit a variable contrast and background. Filament positions are determined by filtering with a point spread function that incorporates the local symmetry of the lattice. Filament orientations are determined by correlation with a template that incorporates the salient filament characteristics, and the orientations are classified using a Gaussian mixture model. The precision of the technique is assessed by application to a variety of micrographs and comparison with manual classification of the orientations. The system provides a convenient, robust, and rapid means of analysing micrographs containing many filaments to study the distribution of filament orientations.