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
Released Data Link
Team
Principal Investigator
Team Members