Acquisition and reconstruction of brain tissue using knife-edge scanning microscopy

A fast method for gathering large-scale data sets through the serial sectioning of brain tissue is described. These data sets are retrieved using knife-edge scanning microscopy, a new technique developed in the Brain Networks Laboratory at Texas A&M University. This technique allows the imaging of tissue as it is cut by an ultramicrotome. In this thesis the development of a knife-edge scanner is discussed as well as the scanning techniques used to retrieve high-resolution data sets. Problems in knife-edge scanning microscopy, such as illumination, knife chatter, and focusing are discussed. Techniques are also shown to reduce these problems so that serial sections of tissue can be sampled at resolutions that are high enough to allow reconstruction of neurons at the cellular level.