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because of its ease of use, excellent image quality, and excellent throughput, serial block-face scanning electron microscopy (sbf-sem) has become the tool of choice for large volume 3d ultrastructural imaging of the nervous system [14].
and although the method is still time consuming, the time investment is only a fraction of the time investment to obtain data by serial sectioning and standard electron microscopy methods. besides its speed, sbf-sem additionally offers excellent preservation of the ultrastructure. this makes the method highly suitable for neuroanatomy [8], [15], and it has been applied to the study of various neuronal cell types in the brain of both invertebrate and vertebrate species (e.g. [9] [11], [16] ). in addition, this method allows for the imaging of any type of tissue and can even be performed at low voltage, which opens the possibility of examining biological samples at the cellular level at very low temperatures or while the biological sample is living.