The 2010 Gordon Research Conference on Three-Dimensional Electron Microscopy will highlight advances in three-dimensional structure determination of cells and biological macromolecules by electron tomography, 2D electron crystallography and single particle cryo-EM. The conference will focus on recent methodological advances in electron microscopic hardware and detector development as well as new image analysis/software methods for 3D structure determination. Current limitations in the available technology will be discussed in the highly interactive and collegial atmosphere of this Gordon Conference. The conference includes a mix of long and short presentations and poster sessions that will foster stimulating interactions between participants, comprising both senior and junior investigators, as well as postdoctoral fellows and graduate students. There will be invited plenary lectures, contributed poster presentations and generous amounts for discussions as well as informal gatherings in the evenings in the stimulating Tuscany ambiance.