Scanning electron microscopy (SEM) is used to observe the surface structure of an object by irradiating an electron beam onto the sample and detecting the reflected and emitted electrons. Because of its large depth of focus, SEM can provide the three-dimensional structure of small surfaces that cannot be observed using an optical microscope. Furthermore, the cross-sectional structure of the tissue can be observed by freeze-cracking. Observing the ultrastructure of organisms that contain large amounts of water in their bodies while maintaining high resolution is challenging; however, this has recently become possible. Here, we explain the fixation and freeze-cracking method for mouse brain samples.
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