2015-09-02 OBS 2014-08-19 RCSB RCSB 2014-09-24 2015-09-02 2015-09-02
Authors will resubmit map at a later date.
Imaging proteins and cells in liquid solution at nanometer resolution by electron microscopy Ren G, Peng B, Zhang L, Lei D, Lu Z, Wong E, Zhang M, Rames MJ, Liu J, De Yoreo JJ, Zhang J Liquid-cell transmission electron microscopy, protein structure in liquid solution, bacteria structure in growth medium, virus infecting mammalian cell Ren G, Peng B, Zhang L, Lei D, Lu Z, Wong E, Zhang M, Rames MJ, Liu J, De Yoreo JJ, Zhang J Imaging proteins and cells in liquid solution at nanometer resolution by electron microscopy To Be Published Zhang L, Ren G IPET and FETR: Experimental Approach for Studying Molecular Structure Dynamics by Cryo-Electron Tomography of a Single-Molecule Structure PLOS ONE 7 e30249 e30249 2012 22291925 doi:10.1371/journal.pone.0030249 emd_6039.map.gz Image stored as Reals 360 360 360 -180 -180 -180 179 179 179 360 360 360 3859.2002 3859.2002 3859.2002 90.0 90.0 90.0 X Y Z -1.51660800 1.04050040 0.00636720 0.07338254 1
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10.72 10.72 10.72 3D reconstruction of an HIV-like particle landing on the membrane surface of a HeLa cell in liquid solution. The particle was imaged in liquid solution and the 3D reconstruction was performed using IPET (Individual-Particle Electron Tomography).
emd_6039.tif
2 An HIV-like virus in the "landing" stage of infecting a HeLa cell. Images were acquired in liquid solution by electron microscopy, and the 3D reconstruction was performed using IPET. a single HIV-like virus bound to the surface membrane of a HeLa cell
The lentivirus is a replication-incompetent HIV that was generated using a third-generation packaging system for the expression of non-functional short hairpin RNA.
virus Lentivirus replication-incompetent HIV
A HIV-like virus infecting a HeLa cell surface.
Lentivirus replication-incompetent HIV true false VIRUS-LIKE PARTICLE OTHER VERTEBRATES Homo sapiens cellular-component surface membrane Homo sapiens human false HeLa cell NONE NONE Objective lens astigmatism was corrected at 125,000 times magnification. LAB6 26 Zeiss Libra 120 PLUS BRIGHT FIELD 293 11000 FLOOD BEAM 296 Normal room-temperature specimen holder
Low-dose illumination
GATAN ULTRASCAN 4000 (4k x 4k) 2.2 -36 13988 66 295 ZEISS LIBRA120PLUS 15-NOV-2010 OTHER 120 10000 0-20 103 16 No stain, in liquid solution, at room temperature cell Two 200 mesh copper TEM grids pre-coated with a thin layer of Formvar film, no glow discharge
MEM medium (containing 10% FBS with 5% CO2)
7.4 tomography Individual-Particle Electron Tomography (IPET) IPET, Spider, EMAN, TOMOCTF, IMOD TOMOCTF
The resolution was defined based on intra-Fourier shell correlation, in which the aligned images were split into two halves (odd index or even index) to generate two 3D reconstructions for computing FSC curves against spatial frequency shells in Fourier space. The frequency at which the FSC curve falls to a value of 0.5 was used to represent the resolution of the IPET 3D density map.
73 FSC 0.5, semi-independent C1 1 103
Whole micrographs were initially aligned using the IMOD software package. CTF was then corrected using TOMOCTF. Small areas within the images (square windows of 128 to 360 pixels, i.e., ~137 to 386 nm) containing the targeted virus were tracked, selected, windowed, aligned, and refined using individual-particle electron tomography (IPET) software.