Evaluation of imaging plates as recording medium for images of negatively stained single particles and electron diffraction patterns of two-dimensional crystals.
We evaluated imaging plates (IPs) and the DITABIS Micron scanner for their use in recording images of negatively stained single-particle specimens and electron diffraction patterns of two-dimensional crystals. We first established the optimal imaging and read-out conditions for images of negatively stained single-particle specimens using the signal-to-noise ratio of the images as the evaluation criterion. We found that images were best recorded on IPs at a magnification of 67,000x, read out with a gain setting of 20,000 and a laser power setting of 30% with subsequent binning over 2 x 2 pixels. Our results show that for images of negatively stained specimens, for which the resolution is limited to approximately 20 A, IPs are a good alternative to EM film. We also compared IPs with a 2K x 2K Gatan charge-coupled device (CCD) camera for their use in recording electron diffraction patterns of sugar-embedded two-dimensional crystals. Diffraction patterns of aquaporin-0 recorded on IPs and with the CCD camera showed reflections beyond 3 A and had similar R(Friedel) as well as R(merge) values. IPs can thus be used to collect diffraction patterns, but CCD cameras are more convenient and remain the best option for recording electron diffraction patterns.
Li
Zongli
Z
Department of Cell Biology, Howard Hughes Medical Institute, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
Hite
Richard K
RK
Cheng
Yifan
Y
Walz
Thomas
T
D020346:Aquaporins / Q000648:ultrastructure; D003461:Crystallography / Q000295:instrumentation / Q000379:methods; D005136:Eye Proteins / Q000648:ultrastructure; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation*; D046529:Microscopy, Electron, Transmission / Q000295:instrumentation* / Q000379:methods; D046912:Multiprotein Complexes / Q000648:ultrastructure; D010781:Photography; D046988:Proteasome Endopeptidase Complex / Q000648:ultrastructure; D011990:Receptors, Transferrin / Q000648:ultrastructure; D014168:Transferrin / Q000648:ultrastructure
D020346:Aquaporins; D005136:Eye Proteins; D046912:Multiprotein Complexes; D011990:Receptors, Transferrin; D014168:Transferrin; C030328:aquaporin 0; D046988:Proteasome Endopeptidase Complex