Reconstructing Plant Cells in 3D by Serial Section Electron Tomography

In micrographs acquired with a transmission electron microscope, 3-dimensional (3D) objects are superimposed onto a 2D screen. This reduction in dimension necessarily leads to a degradation of image resolution. To overcome this problem, 3D microscopy techniques, such as tomography and single particle analysis, have been developed. Tomography has been used to visualize cells in 3D, and single particle analysis has been used to investigate macromolecules and viral particles. In this chapter we will describe how we have collected tilting series micrographs from plant cells and how we have reconstructed the cellular volumes using dual axis electron tomography.