Automated Cryo Electron Tomography Data Collection

Electron tomography entails the three-dimensional reconstruction of an object from its projections, which are acquired using a transmission electron microscope (TEM). This chapter describes the process of collecting consecutive images of the tilted specimen efficiently in theory and practice. Firstly, the concept of a TEM and its main components are described prior to introducing the technical means to maintain an area of interest in the field of view during tilting. Then different tilt schemes are introduced as well as current approaches to maximize throughput and obtained signal per electron dose. A more detailed description of the TEM setup and different sample properties then allows a hands-on description of tilt series collection and its pitfalls. Lastly, technical innovations are discussed that may improve the optical quality of tiltseries in the coming years. In summary, this chapter provides the conceptual and practical framework for state-of-the-art data acquisition.