THEME 1 MOLECULAR IMAGING USING XFEL

Molecular imaging of protein dynamics using megahertz X-ray sources: The illustration in the foreground is of a microfluidic mixing device developed at the imaging CoE that enable capture of sub-millisecond protein dynamics at the XFEL. The background image shows the structure of the first ever membrane protein to be solved at the European XFEL. The goal is to create molecular movies of the light-induced changes to the electron density that occur during photosynthesis.

“Through our partnership with the European XFEL and DESY we are now able to collect atomic scale molecular movies
faster than ever before”

Capturing the real-time, atomic scale, dynamics of molecules in crystals and in solution offers major opportunities for drug development. Historically, one of the biggest challenges facing researchers has been collecting sufficient data to enable a complete movie of molecular motion to be. Megahertz Free Electron Laser sources addressed this problem, and in 2019, the first molecular movies using this new source were published. Imaging CoE researchers have played an essential role in driving this work and, so far, are the only scientists from Australian scientists who have had the opportunity to participate in these ground-breaking experiments.

Photo of Professors Brian Abbey and Keith Nugent

PROF. BRIAN ABBEY
PROF. KEITH NUGENT

AT A GLANCE

The advent of ultra-bright, X-ray Free Electron Laser (XFEL) sources is driving fundamental advances in our ability to study molecular structure at the atomic scale. However, as the field of time-resolved molecular movies at the XFEL has progressed, it has become increasingly clear that some of the biggest challenges when imaging atomic scale dynamics are related to building up sufficient statistics. Due to the random orientation of the crystal as it arrives at the XFEL beam, it typically takes 1000s of diffraction patterns to image the 3D structure of a static molecule using serial crystallography. To make a molecular movie one needs a high-resolution 3D structure of the molecule corresponding to each ‘frame’ of the move. Creating atomic scale molecular movies is a 4D problem, requiring orders of magnitude with more data than standard crystallography. This is why the Imaging CoE’s access to, and partnership with, the European XFEL is so critical, as it is currently the only X-ray source in the world able to collect 10’s of thousands of images of molecules per second.

In 2019, the Imaging CoE collaborated on the first publication which described how megahertz XFEL sources could be exploited for faster molecular imaging (Yefanov et al, Structural Dynamics, 2019) and played a leading role in solving the first membrane protein structure at the European XFEL (Gisriel et al., Nature Communications, 2019).

In 2019, the joint La Trobe – European XFEL PhD agreement was signed with the first PhD students (Jaydeep Patel and Trey Guest) to join the program. These students are working on projects that will directly contribute towards the global research effort in atomic scale molecular imaging. The Imaging CoE’s work in structure determination using XFELs was highlighted in the international scientific press (e.g. Science Daily and Eureka Alert). Other recent successes include, the continued recognition for the La Trobe groups commercialisation of their molecular imaging platform with the 2019 award of Australia’s largest medical technology prize: Medtech’s Got Talent. The La Trobe Imaging CoE group won $30,000 (undiluted funds) out of more than 100 entrants for being the “most investible team”.

ACTIVITY PLAN

  1. Continue to support external users of Australia’s first serial crystallography facility that was designed and developed by the Imaging CoE in partnership with ANSTO.
  2. Continue data analysis of molecular dynamics diffraction data collected from immunological targets at the AS SAXS/WAXS beamline.
  3. In collaboration with RMIT and the University of Melbourne, La Trobe University will continue to lead experiments applying fluctuation microscopy to study protein-protein interactions.
  4. In collaboration with the theory group at Melbourne, continue research into understanding the fundamental physics governing electronic damage at XFELs.
  5. Aim to spin out molecular imaging technology with support from both university and industry partners.

HIGHLIGHT

Serial crystallography arrives in Australia

In 2019, the first papers were published using the newly developed serial crystallography facility at the Australian Synchrotron. The facility is a joint research project led by La Trobe University, in collaboration with ANSTO and Monash University. It is based on the $3.2M Eiger Detector at MX2, which was supported by a grant from the Cancer Council Victoria.

The Australian Synchrotron serial crystallography facility using the Eiger Detector enables over 100 diffraction patterns to be collected each second from protein crystals which are continually streamed through the X-ray focus. This rate of data collection means that structural data can be collected faster and more efficiently, and samples can be delivered in-situ at room temperature within a viscous stream of lipid.

The development of serial crystallography has been supported by ANSTO’s Professor Andrew Peele, a Partner Investigator in the Imaging CoE.

“Serial crystallography is an important part of the roadmap for the future development of molecular imaging capabilities at the Australian Synchrotron,” said Professor Andrew Peele.

“By building up a user-friendly platform that is accessible to a wide range of different researchers, we ensure a great legacy and lasting impact for this ARC Centre of Excellence,” Chief Investigator Prof. Brian Abbey said.

The serial crystallography facility that was developed as a joint ARC Imaging CoE – ANSTO collaboration led by La Trobe University.

ACHIEVEMENT

Translating molecular imaging technology

In early 2019, Chief Investigator Prof. Brian Abbey and CoE Senior Research Fellow Dr Eugeniu Balaur were the overall winners of the national Medtech’s Got Talent (MTGT) prize. As part of the competition, which was held in Sydney, they pitched their technology and then answered questions from an expert panel. The panelists included venture capitalists and medical technology experts.

The competition began in 2018, and they passed through a series of three pitching events in both Melbourne and Sydney. Prof. Abbey and Dr Balaur won $30,000 in undiluted funds, and the technology resulted in four new PCT applications filed in 2019.

Following the win, they both spent 2019 pursuing a commercial journey and were invited to participate in The Actuator, an Australian medical technology accelerator program. This six-month program provided training, expertise and facilities to enable them to develop and begin a startup.

The La Trobe team are now aiming to spin-out their technology from the University, and have identified a range of applications for their platform technology, which spans medical, agricultural, as well as the materials sciences.

Imaging CoE researcher and co-inventor Dr Eugeniu Balaur.