Electron microscopy coupled with quick freezing
Transmission electron microscopy is a unique means of providing high resolution real-space images of biological ultrastructure and thus has greatly contributed to the detailed structural studies of muscle at various levels, from the morphology of tissues or cells to the submolecular structure of each contractile protein. However, the fixation and/or the staining procedures, which are required to visualize the fine structural details under the microscope, include time-consuming chemical processes in an aqueous phase, raising the possibility that the final images might include certain changes from the original native and thus physiological structures. The primary aim of this chapter is to give a brief explanation and critical evaluation of each technique used in quick-freeze electron microscopy of muscle and contractile proteins. Though there is a large literature on studies made with such techniques, the results of each study are not discussed here, except for a brief summary on those studies related to the most intriguing structural change of myosin cross-bridges during muscle contraction.
Keywords: electron microscopy, real-space images, submolecular structure, chemical processes, quick freezing, muscle contraction
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