Trehalose embedding technique for high-resolution electron crystallography: application to structural study on bacteriorhodopsin

Trehalose was compared with glucose in its use as an embedding medium for the preparation of two-dimensional crystals for electron crystallographic structure analysis. Purple membranes (two-dimensional crystals of bacteriorhodopsin) embedded in either sugar were studied in a 'partially hydrated' state as well as after 'drying' of the specimen. Diffraction patterns of the four conditions, all showing spots to a resolution better than 3.0 A, were analysed and merged. In the case of glucose embedded specimens, the merging R-factor obtained from the diffraction patterns of dried specimens was significantly better than the one obtained from partially hydrated specimens. In the case of trehalose embedding, the merging R-factor for partially hydrated specimens was significantly better than the dried one. The trehalose embedded, partially hydrated specimens gave the best merging R-factor in all four cases.