Glacios cryo-electron microscope for single particle analysis and electron tomography of proteins, complexes and fibrillar assemblies

Electron microscopy has been used for decades to provide details of cell and tissue structures and to image particles that could not be seen by light microscopes, such as viruses. However, in recent years electron microscopy has undergone a "Resolution Revolution" with a combination of improvements in freezing samples (cryoEM) along with software and hardware advances. The details that can now be observed from cryoEM data include the structure of proteins to atomic resolution, the level where all atoms in the molecule can be seen. The "Resolution Revolution" has led to many new structures being determined in recent years and was acknowledged in the award of the 2017 Nobel Prize in Chemistry for these developments. Importantly, proteins that were previously too difficult to determine their atomic resolution structure are now being imaged for the first time using cryoEM data.

At the University of Manchester, we have many projects supported by BBSRC funding that require access to cryoEM. However, our current microscope, the Polara, is out-dated and no longer supported by the manufacturer. Therefore, this application is to provide a state-of-the-art Glacios cryo-electron microscope to support the research of a very productive team of scientists at the University of Manchester. The Glacios cryoEM is a step-change in microscope technology which would allow us to collect data at a much faster rate, due to advances in automation, which means more images can be collected each day. The Glacios is also much more stable which will allow us to collect higher resolution images. In addition, the Glacios has compatible cryoEM sample handling with other instruments, including those in the national electron bioimaging centre, which means samples can be transferred between microscopes for data collection.

The research at Manchester that needs access to a new cryoEM covers a broad range of science that is in alignment with BBSRC's strategic priorities. Projects include the analysis of extracellular matrix proteins involved in cell signalling, tissue strength and inflammation; determining the structures of membrane proteins responsible for multidrug resistance and kidney function; investigation of enzyme mechanism for biological catalysis; understanding the processes involved in synthesising proteins, and analysing virus-like particles in the development of new vaccines. These studies will benefit enormously from a new cryoEM and the University of Manchester recognises this by offering major financial support including 35% of the cost of the Glacios, plus a purpose-built room in our Electron Microscopy Facility and an expert experimental officer who will operate the microscope and train new users.

Cryo-electron microscopy (cryoEM) and cryo-electron tomography are state-of-the-art-technologies used to address fundamental questions in biology. The "resolution revolution" has not only transformed the level of detail found in cryoEM structures, but has also broadened the range of proteins and complexes amenable to structure determination. Together this has resulted in an exponential growth in the number of structures solved by cryoEM and includes previously intractable targets such as extracellular matrix and membrane proteins. This application is to provide a state-of-the-art cryo-electron microscope to support the ongoing research of a very productive team of scientists at the University of Manchester. The requested ThermoScientific Glacios cryo-electron microscope will replace a 12yr old Polara EM, which is now outdated and no longer supported by the manufacturer. The Glacios cryoEM represents a step-change in microscope technology, both in terms of reliability and compatibility with other imaging modalities, and would bring enhanced cryoEM capabilities to Manchester. The cutting edge research requiring access to the new cryoEM covers a broad range of biological sciences, from: supramolecular analysis of extracellular matrix components involved in signalling, inflammation and mucus rheology; integral membrane proteins responsible for multidrug resistance and kidney function; investigation of mechanistic questions in biological catalysis; intracellular proteins involved in protein trafficking, vesicle sorting and control of cellular stress, and structural vaccinology with virus-like particles. These studies will gain enormous benefit from the new cryoEM instrument and the University recognises this by offering major support including 35% of the capital cost, plus key infrastructure for its installation and continued staff support for its optimum use.

The requested cryo-electron microscope will support the research of a large number of researchers whose projects address important biological, medical and physical questions. CryoEM is of great importance to UK science and industry, as clearly described in the BBSRC Strategic Review of Bioimaging. It falls under the remit of UKRI Technology Touching Life, and closely aligns with the drivers for BBSRC Exploiting New Ways of Working that states: "strengthening of the skills base is required in order to embed the latest equipment in facilities, and enable multidisciplinary research". This cutting-edge technology will substantially enhance the delivery of BBSRC-funded world-class bioscience by researchers at the University of Manchester.

There is a wide range of direct and indirect beneficiaries:
(1) Biotechnology and biosciences. CryoEM data contributes to research that will reach a broad audience across disciplines, including extracellular matrix research, molecular cell biology, membrane protein structure and function, biotechnology and biological enzymology. Data from the Glacios will be deposited in appropriate databanks, having a direct effect on academic research with immediate benefits and long-term impact.
(2) Industry. We will engage with local companies to offer the Glacios as a cryoEM instrument in the North West. As an example, Allergan, Manchester BIOGEL and Peak Proteins have provided letters of support (attached) indicating their desire to use such an instrument for quality control assays and grid screening prior to data collection at eBiC. Thus, there will be an impact (over the medium to long term) on local industry success and revenue.
(3) Staff working on cryoEM projects. Researchers will develop skills in a new sought-after technique, i.e. single particle cryoEM analysis of proteins and complexes, thereby enhancing their research careers. Indeed, this aligns with the BBSRC's strategic plan in maintaining strength and capabilities in core underpinning disciplines such as structural biology, so fulfilling a core BBSRC goal. These individuals will further develop communication, problem solving and analytical skills, which will be useful in their professional development (regardless of sector). The impact will be direct, evident in the short-term and with a lasting legacy.
(4) General public. Structures and 3D printed models generated from cryoEM data are colourful, intuitive, and attractive, and make science more accessible. They are useful for educating the public, and particularly capture the imagination of children, who will be encouraged to engage with science through the school sessions run by the applicants e.g. the annual FBMH Community Open Day and Manchester Science Festival. The Wellcome Centre for Cell-Matrix Research has a dedicated public engagement programme manager to facilitate the delivery of such events. The breadth of science covered by this proposal will demonstrate how disciplines can be integrated to deliver tangible benefits for society, from understanding and treating disease, to the development of new vaccines and bio-industrial processes. The general public (of the UK and beyond) will also benefit indirectly from improved quality of life, as our discoveries are translated into new healthcare solutions or policy guidance. We will report research breakthroughs in the local, national and international press via the Faculty's Media Relations Office. Thus, there will be both direct and indirect impacts on the General Public ranging from short- to long-term.