Methods

Molecular electron microscopy, the use of electron microscopy to determine the structure of proteins and macromolecular complex, is still a relatively young technique and there is still much room for advancement of the methods used in this field. Our main interest concerns the use of electron microscopy to address specific biological questions. We intend to implement all the major electron microscopic techniques in our laboratory to take full advantage of the versatility provided by electron microscopes. Electron crystallography and single particle electron microscopy are techniques already well established in our laboratory. More recently we have also begun to establish electron tomography in our laboratory. While we will use electron tomography in the traditional way to obtain 3D reconstructions of thin sections, we will also attempt to employ electron tomography to generate initial 3D models from vitrified particles. The goal is to use these initial models to align images of vitrified samples taken from heterogeneous specimens. All research using the electron tomographic approach will be directed by Yifan Cheng. While we are not focused on the development of methods, we do work on improving techniques if currently available methodology is incapable or not well suited to provide the answer we seek to a biological question. A typical example is the adaptation of methods routinely used in X-ray crystallography to electron crystallography. In this long-term project we rely strongly on Stephen Harrison’s comprehensive expertise in X-ray crystallographic techniques. Another example is our work on adapting the lipid monolayer technique to the preparation of single particle specimens.

Electron Crystallography

Single Particle EM