DOI: 10.22002/8rbeq-svh12
title: H.pylori 26695 WT/AGS

description: grid box: YWC081
description: <strong>Tilt Series Date:</strong> 2014-02-14</p>
<strong>Data Taken By:</strong> Yi-Wei Chang</p>
<strong>Species / Specimen:</strong> Helicobacter pylori</p>
<strong>Strain:</strong> 26695</p>
<strong>Tilt Series Settings:</strong>  Single Axis, tilt range: (-60.0°, 60.0°), step: 1.0°, constant angular increment, dosage: 205.0 eV/Å²,  defocus: -10.0 μm,  magnification: 22500x. </p>
<strong>Microscope:</strong> Caltech Polara</p>
<strong>Acquisition Software:</strong> UCSFTomo</p>
<strong>Upload Method:</strong> pipeline</p>
<strong>Processing Software Used:</strong> Raptor</p>
<strong>Collaborators and Roles:</strong> H.pylori & AGS cells from Carrie Shaffer</p>
<strong>Purification / Growth Conditions / Treatment:</strong> H.pylori cells grown in BLBB + 5% heat inactivated fetal bovine serum at 37 degrees in 5% CO2 chamber for O/N, OD600=~0.5. AGS cells cultivated on QF Au finder grid for 1 day in RPMI + 10% FBS. Infect by pylori for 4 hr.</p>
<strong>Sample Preparation:</strong> Vitrobot settings: 100% humidity, 37 C, 3µl BSA treated 20 nm gold (100 ul gold+ 25 ul BSA, pellets from 6 tubes merged into ~30 ul final solution). Manual blot by Whatman 40 filter paper for 3 sec. Ethane/propane mix.</p>
  ycw2014-02-14-3_slicer3660.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  Another view of big tube with rows of small tubes on it, possibly T4SS related, projecting from Hp cell. ycw2014-02-14-3_slicer3661.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  Crystalline lattice (chemoreceptor array?) and filament bundles ycw2014-02-14-3_slicer3662.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  Broken tube projecting from Hp cell. Possibly T4SS related. ycw2014-02-14-3_slicer3663.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  Can see subunits of filament bundles. ycw2014-02-14-3_slicer3664.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  Tube projecting from Hp cell. Possibly T4SS related. ycw2014-02-14-3_slicer3665.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  Big tube projecting from Hp cell. Protein subunits visible.  Possibly T4SS related. ycw2014-02-14-3_slicer3666.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  View down small tubes that decorate big tube projecting from Hp cell. Possibly T4SS related. ycw2014-02-14-3_slicer3667.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  Tube projecting from Hp cell. Possibly T4SS related. ycw2014-02-14-3_slicer3668.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  Rings of 35nm diameter (arrows) and crystalline array. ycw2014-02-14-3_slicer3669.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  Round structures (pores?) with 15nm diameter in outer membrane of Hp. ycw2014-02-14-3_slicer3670.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  Filament bundles ycw2014-02-14-3_slicer3671.jpg: Helicobacter pylori 26695 WT co-cultured with AGS cells.  Hp cell is lysing. .   .  50nm ring in membrane is same width as big tubes, but doesn't have any z-height.
description: Files available via S3 at https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/ycw2014-02-14-3/rawdata/</p>20140214_YWC081_10003.mrc,  Tilt Series (Pixel Size 0.48 nm), 3.4 GB  
        
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        20140214_YWC081_10003_part121_1.rec,  Reconstruction (Pixel Size 1.92 nm), 712.7 MB  
        
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