DOI: 10.22002/3bxmg-r3945
title: Lp02_2nd

description: 2nd time collection: Jan 2016. Defocus -6.Lp02 is thymidine auxotroph of the parental strain Lp01 originating from Philadelphia-1 strain. Cells were grown in AYE liquid culture with following supplements: ferric nitrate, cysteine hydrochloride and thymidine. Scrape cells off a plate, dissolve in the media, adjust the cell density to 0.6 and make a dilution series. Start O/N culture with all of them and freeze cells next day from the one in early stationary phase that is OD ~ 3.0
description: <strong>Tilt Series Date:</strong> 2016-01-15</p>
<strong>Data Taken By:</strong> Debnath Ghosal</p>
<strong>Species / Specimen:</strong> Legionella pneumophila</p>
<strong>Strain:</strong> Philadelphia-1_Lp02</p>
<strong>Tilt Series Settings:</strong>  Single Axis, tilt range: (-60.0°, 60.0°), step: 1.50°, constant angular increment, dosage: 100.0 eV/Å²,  defocus: -6.0 μm,  magnification: 26000x. </p>
<strong>Microscope:</strong> Janelia Titan</p>
<strong>Acquisition Software:</strong> UCSFTomo</p>
<strong>Upload Method:</strong> reupload_downloaded</p>
<strong>Processing Software Used:</strong> TOMO3D</p>
<strong>Collaborators and Roles:</strong> Strain sent to us by Joseph P. Vogel, Washington University School of Medicine. Data collected and processed by Debnath Ghosal. This data was collected at the Janelia Krios.</p>
<strong>Purification / Growth Conditions / Treatment:</strong> Lp02 is thymidine auxotroph of the parental strain Lp01 originating from Philadelphia-1 strain. Cells were grown in AYE liquid culture with following supplements: ferric nitrate, cysteine hydrochloride and thymidine. Scrape cells off a plate, dissolve in the media, adjust the cell density to 0.6 and make a dilution series. Start O/N culture with all of them and freeze cells next day from the one in early stationary phase that is OD ~ 3.0. Added gold before freezing.</p>
<strong>Sample Preparation:</strong> Grids were stored in Janelia from previous imaging. See previous Lp02 sample prep. Vitrobot (MK iv) settings: 100% humidity, 4µl sample, Blot force 6, blot time 3 sec. Wait time 0 sec. Drain time 1 sec. Ethane/propane mix. Grid: Quantifoil Cu 2/2 200. Add gold to cells prior to freezing: 1ul of gold to 4 ul of cells, freeze.</p>
  dga2016-01-15-88_slicer16682.jpg: Flagella like filament wrapping the cell and this filament is in the periplasm. dga2016-01-15-88_slicer16683.jpg: Flagella like filament wrapping the cell and this filament is in the periplasm. dga2016-01-15-88_slicer16684.jpg: Flagella like filament wrapping the cell and this filament is in the periplasm. dga2016-01-15-88_slicer16685.jpg: Flagella like filament wrapping the cell and this filament is in the periplasm. dga2016-01-15-88_slicer16686.jpg: Flagella like filament wrapping the cell and this filament is in the periplasm.
description: Files available via S3 at https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2016-01-15-88/rawdata/</p>atlas_area_a0002.mrc,  Tilt Series (Pixel Size 0.42 nm), 2.2 GB  
        
<a role="button" class="ui compact mini button" href="https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2016-01-15-88/rawdata/atlas_area_a0002.mrc"
        > <i class="download icon"></i> Download </a></p>

        atlas_area_a0002_SIRT_1k.rec,  Reconstruction (Pixel Size 0.0 nm), 1.0 GB  
        
<a role="button" class="ui compact mini button" href="https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2016-01-15-88/3dimage_108349/atlas_area_a0002_SIRT_1k.rec"
        > <i class="download icon"></i> Download </a></p>

        atlas_area_a0002.tlt,  648.0 B  
        
<a role="button" class="ui compact mini button" href="https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2016-01-15-88/file_24901/atlas_area_a0002.tlt"
        > <i class="download icon"></i> Download </a></p>

        T2SSK.mod,  1.0 kB  
        
<a role="button" class="ui compact mini button" href="https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2016-01-15-88/file_24902/T2SSK.mod"
        > <i class="download icon"></i> Download </a></p>

        T2SSK_2k.mod,  1.0 kB  
        
<a role="button" class="ui compact mini button" href="https://sdsc.osn.xsede.org/ini210004tommorrell/tomography_archive/dga2016-01-15-88/file_24903/T2SSK_2k.mod"
        > <i class="download icon"></i> Download </a></p>