Poster Presentation The 44th Lorne Conference on Protein Structure and Function 2019

New Ideas in Customer-centric CryoEM Automated Collection, Workflows, and Processing for High-resolution Protein Structures (#293)

Simon HJ Brown 1 2 , Claudia S Kielkopf 1 3 , Gökhan Tolun 1 3 , Lance Wilson 4 , James C Bouwer 1 2 , Wojtek J Goscinski 4 , Antoine van Oijen 1 2 3 , Nicholas Dixon 1 2 3
  1. School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW, Australia
  2. Molecular Horizons, University of Wollongong, Wollongong, NSW, Australia
  3. Illawarra Health and Medical Research Institute, Wollongong, NSW, Australia
  4. MASSIVE, Monash University, Clayton, Vic, Australia

Cryo-electron microscopy (cryo-EM) has undergone a revolution over the past decade, with improvements in imaging detector systems, hardware, automation and data analysis rapidly pushing the limits of resolution to below 2 angstroms and allowing smaller (<60 kD) proteins to be reconstructed. The University of Wollongong (UOW) has established cryo-EM as a foundational capability in the Molecular Horizons Institute, a centre for molecular and life sciences. Fundamental to UOW’s vision has been the acquisition of three FEI electron microscopes: a Tecnai T12, a Talos Arctica and a Titan Krios. This suite of microscopes is integrated with a fully-equipped sample preparation lab, enabling a complete single-particle EM workflow through from negative-stain screening to high-resolution data collection and reconstruction.

UOW has partnered with the MASSIVE High Performance Computing facility (coordinated by Monash University) allowing microscope users to access world-class HPC resources to process cryo-EM data. The UOW Molecular Horizons cryo-EM facility has intentionally been designed as a full-service operation, providing client training and support during sample preparation, grid screening, microscopy and data analysis. Training is conducted both in regular workshops, as well as one-on-one sessions, enabling clients to achieve high levels of proficiency in EM techniques as well as self-sufficiency in microscope operation. In this poster we will present an outline of the current and planned capabilities of the UOW cryo-EM facility, as well as results from recent benchmarking experiments on our Talos Arctica and Titan Krios instruments.