Charge-coupled device (CCD) cameras have found wide application in electron microscopy. They provide much faster availability of image and diffraction data than photographic film, and this improvement in turn around time has allowed a tremendous increased in the efficiency of specimen evaluation, microscope operation and automation, and data recording. For some types of work the data collected with a CCD is of higher quality than when recorded on film. However, the CCD performance is seriously compromised when the microscope is operated much above 100 kV, due to the decrease in efficiency of the scintillator, which decreases the signal level, and increased lateral scattering of the electrons within the scintillator, which decreases the resolution. Together these effects can produce a detective quantum efficiency (DQE) which is far below that of film, and indeed too low for work such as low-dose imaging of proteins. The use of intermediate Voltage Electron Microscopes (IVEMs) operating at 300-400 kV is increasing as the many advantages of the higher accelerating voltage become more widely recognized, so the CCD performance on these microscopes is becoming more of a limitation. We plan to overcome the poor performance of CCD cameras on IVEMs by decelerating the electrons before they reach the camera. We will mount the CCD so that it can be floated to 200 kV, so that when the microscope is operated at 300 kV we will obtain the advantages of the IVEM but the camera will perform as well a on a 100 kV microscope. Further improvements will be made to the scintillator to produce even better signal and resolution characteristics than currently available. These adaptations will produce substantial benefits in work ranging from high-resolution structural studies of membrane proteins and viruses to the three dimensional study of cell ultrastructure.
| Zhang, Rui; LaFrance, Benjamin; Nogales, Eva (2018) Separating the effects of nucleotide and EB binding on microtubule structure. Proc Natl Acad Sci U S A 115:E6191-E6200 |
| Nogales, Eva (2018) Cytoskeleton in high resolution. Nat Rev Mol Cell Biol 19:142 |
| Downing, Kenneth H; Glaeser, Robert M (2018) Estimating the effect of finite depth of field in single-particle cryo-EM. Ultramicroscopy 184:94-99 |
| Nogales, Eva (2018) Cryo-EM. Curr Biol 28:R1127-R1128 |
| Sazzed, Salim; Song, Junha; Kovacs, Julio A et al. (2018) Tracing Actin Filament Bundles in Three-Dimensional Electron Tomography Density Maps of Hair Cell Stereocilia. Molecules 23: |
| Kamennaya, Nina A; Zemla, Marcin; Mahoney, Laura et al. (2018) High pCO2-induced exopolysaccharide-rich ballasted aggregates of planktonic cyanobacteria could explain Paleoproterozoic carbon burial. Nat Commun 9:2116 |
| Howes, Stuart C; Geyer, Elisabeth A; LaFrance, Benjamin et al. (2018) Structural and functional differences between porcine brain and budding yeast microtubules. Cell Cycle 17:278-287 |
| Glaeser, Robert M (2018) PROTEINS, INTERFACES, AND CRYO-EM GRIDS. Curr Opin Colloid Interface Sci 34:1-8 |
| Kellogg, Elizabeth H; Hejab, Nisreen M A; Poepsel, Simon et al. (2018) Near-atomic model of microtubule-tau interactions. Science 360:1242-1246 |
| Han, Bong-Gyoon; Watson, Zoe; Cate, Jamie H D et al. (2017) Monolayer-crystal streptavidin support films provide an internal standard of cryo-EM image quality. J Struct Biol 200:307-313 |
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