Development of observation technology with electron microscope of force-generating myosin

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We tried to develop observation technology with electron microscopes of force generating myosin. One necessary technology is to observe the active interaction between a myosin filament and an actin filament during force generation. The actin filament could be, in success, pulled by a magnetic bead, which was linked to the actin covalently. The other is to observe three-dimensional structure of single myosin head and actin filament by electron cryo-microscopy. We developed new three-dimensional reconstruction programs for single particle analysis and electron tomography of frozen-hydrated specimens and obtained highly-resolved 3D map of actin filaments/actomyosin complexes and preliminary 3D maps of single head myosin bound to a single actin filament.