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Visualizing pathways in cells for more accurate biodesign


EMSL Project ID
50188

Abstract

In this Capability Development Focus project, we plan to acquire label-free whole cell three-dimensional (3D) tomograms of the two primary organisms of interest for the FY18 Subcellular SSA thrust. We will additionally, begin developing a workflow for cryogenic sample handling to allow similar datasets to be acquired on cells currently too large for direct TEM analysis at EMSL. Our specific aims are:
1) Acquire cryogenic electron tomography datasets with nanometer spatial resolution from whole cells of the smallest free-living eukaryote (Ostreococcus tauri) and a bacterium overlapping with the Intercellular SSA (Arthrobacter chlorophenolicus). Both will be imaged under various nitrogen limited conditions.
2) Adapt dual beam Focused Ion Beam/Scanning Electron Microscopy (FIB-SEM) capabilities at EMSL to allow cryogenic lift-outs, lamellar thinning and serial section slice-and-view from large samples.
These datasets will help launch subcellular and cellular modeling and visual proteomics projects for FY19 and beyond. It will also extend cryo-electron tomography capabilities at EMSL which can immediately benefit EMSL user projects for FY19.

Project Details

Start Date
2018-03-22
End Date
2018-09-30
Status
Closed

Team

Principal Investigator

James Evans
Institution
Environmental Molecular Sciences Laboratory

Team Members

Chuck Smallwood
Institution
Environmental Molecular Sciences Laboratory

Trevor Moser
Institution
Environmental Molecular Sciences Laboratory

Daniel Perea
Institution
Environmental Molecular Sciences Laboratory

Tamas Varga
Institution
Environmental Molecular Sciences Laboratory