1 / 7

PCAP EM Project: Whole-Cell Electron Tomography

PCAP EM Project: Whole-Cell Electron Tomography. Kenneth H. Downing, David Andrew Ball. The Goal Mapping (large) macromolecular complexes in tomograms of intact cells The Requirement Resolution sufficient to see and to discriminate among complexes - 20 - 50 Å The Problem

lucine
Download Presentation

PCAP EM Project: Whole-Cell Electron Tomography

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. PCAP EM Project:Whole-Cell Electron Tomography Kenneth H. Downing, David Andrew Ball

  2. The Goal Mapping (large) macromolecular complexes in tomograms of intact cells The Requirement Resolution sufficient to see and to discriminate among complexes - 20 - 50 Å The Problem Resolution decreases as specimen thickness increases; DvH is a bit thick…

  3. Several possible solutions: Specimen flattening during preparation ‘Metabolic’ thinning Dual-axis tilt series Bursting cells (electroporation, pressure) Mechanical thinning Cryo-sectioning * Other (thinner) cells ** Better images

  4. Focused Ion Beam Milling

  5. Resin section

  6. Pyruvate grown DvH in 100ml flasks Pyruvate grown DvH overview Pyruvate grown DvH Sulfate grown DvH -Sulfate grown DvH average 2D width = ~700-800 nm -Pyruvate grown DvH average 2D width = ~600-650 nm

  7. Pyruvate grown DvH in fermentor

More Related