Sixth International Congress on Electron Tomography

Wim Voorhout, Cellular Biology Solutions

Electron Microscopy has become a very powerful technique to unravel the ultra structural details of cells and macromolecular complexes in three dimensions - down to the subnanometer level. New developments, both in hardware and software, have driven Single Particle Analysis and Electron Tomography into routine approaches obtain this 3D information. Single Particle Analysis can provide high resolution information on viruses, macromolecular machines and proteins and to resolve their 3D structure up to a few angstroms. Electron tomography will provide information on the cellular context of the macromolecular machines and the cellular architecture of parts of the cell.

The desire to understand the 3D relationships of mesoscale hierarchies has led to the development of advanced microscopy techniques. A focused ion beam (FIB) can be used as an alternative for serial section TEM, where the FIB acts as a nano scalpel to allow ultrathin sections down to 5-10 nm and revealing the internal microstructure in a site-specific manner. Scanning Electron Microscopy (SEM) and FIB technologies have now been brought together into a single instrument representing a powerful combination for the study of biological specimens and soft materials. We demonstrate the use of FIB-SEM to study 3D relationships of cellular structure volumes up to several cells.

- Break and Discussion -

Latest Developments Electron Tomography Data Analysis

Remco Schoenmakers, Senior Software Scientist

FEI, as a total solution provider, is covering the electron tomography workflow from sample preparation (Vitrobot), through acquisition (Xplore3D), alignment and reconstruction (Inspect3D), analysis through 3D template matching and sub-tomogram averaging (ARGOS), and visualization (ResolveRT). During this workshop we will focus on the data analysis step and discuss the algorithms, features and highlights of Inspect3D and ARGOS. The workshop will include live previews of the development versions of Inspect3D and ARGOS.

Cowpea Mosaic Virus (CPMV) with x-ray coordinate

Tubes of gp23 from bacteriophage T4

Template matching of vitrified KLH structure