A facility for 3D cellular imaging that bridges light and electron microscopy

In biology, seeing is believing. To find out what is happening inside cells, biologists use microscopes. This equipment comes in different varieties. The two main microscopes biologists use are light microscopes and electron microscopes. With light microscopes, we can image living cells and watch many different proteins going about their job. Using electron microscopes, we can image much smaller things, however, we can only look at fixed (dead) cells and also seeing proteins is difficult. This money from the BBSRC will help us to buy two of the very latest microscopes. We will put these microscopes together with other machines to make it possible for scientists to see living cells and proteins and then see the same cell and the same proteins in the electron microscope. The special thing about these new microscopes is that they can take pictures and movies in 3D, so we can get all the information out of the cells that we are looking at. The equipment will be based at University of Warwick. We have lots of biologists at our University who need these instruments to carry out important research on bacteria, plants and animals, to understand neuroscience, cell division and plant signalling - to name just a few. We will open up the equipment so that other scientists based in the Midlands or even further afield can use it. These scientists may be Biologists or they may be Materials Scientists or Chemists working in Universities or in Industry.

Understanding fundamental processes such as cell division, protein trafficking, synaptic function, and embryo development requires that the fine structure of cells and tissues be imaged in three dimensions and at the highest possible spatiotemporal resolution. Light microscopy (LM) and electron microscopy (EM) are key tools for this and the development of correlative light and electron microscopy (CLEM) approaches has capitalised on the advantages of each technique to allow high resolution imaging of cells at specific stages in their functional cycle. Recent advances in EM and LM hardware bring capabilities that are urgently needed by many scientists at Warwick, within the Midlands and further afield. Our proposal is for LM and EM hardware with which to establish a 3D Cellular Imaging Facility at Warwick. This facility will bridge optical and electron microscopy and provide high resolution 3D bioimaging using both techniques. Users can access the entire 'bridge' workflow, or access experiment-specific subsets of the full workflow. To form the 3D Cellular Imaging Facility, we request funds to purchase: 1) a high-performance TEM with a field emission energy source, direct detection, energy filter and electron tomography capability and 2) a diSPIM light sheet microscope for fast 3D imaging and imaging of thick specimens. This new equipment will allow scientists to obtain 3D images of samples using light and electron microscopy. Light sheet reconstructions of dynamic cells with equivalent spatial resolution in all three dimensions can be combined with 3D electron microscopy imaging using tomography at greatly improved levels of sensitivity to achieve unprecedented accuracy in the correlation of dynamic cell processes with the underlying ultrastructure. The need for this resource is driven by demand from a number of existing BBSRC-funded labs and others at Warwick who urgently need to access this cutting-edge high-resolution capability.

The impact of the new facility will be to fuel several BBSRC Research Priorities. For example, Technology development for the biosciences (Royle, Smith), Data-driven biology (McAinsh) and Sustainably enhancing agricultural production (Cross, Frigerio). In Warwick, we have strong representation in Synthetic biology and Systems approaches to the biosciences, these groups will be able to access the equipment. More broadly, the equipment will Build Partnerships by driving collaborative research with users (see Work Plan) and International and Industrial Partnerships.

The facility will impact upon ACADEMIC SCIENTISTS who will use the new imaging facility to gain high resolution 3D information on samples from a wide range organisms and alter their own research and development activities in light of this. They will be able to achieve unprecedented accuracy in the correlation of dynamic cell processes with the underlying ultrastructure which will contribute to our understanding of cellular mechanisms in projects spanning plant biology, mammalian cell biology and model organisms. The multi-disciplinary environment creased by the facility will further knowledge exchange and learning via the user group meetings.

The insights we uncover will be published in journals of the highest possible calibre, thus sustaining the reputation of the UK as a world leader in scientific enquiry. All investigators on the application have fantastic track records in this regard. Learned bodies, such as the Royal Microscopical Society, Biochemical Society and the British Society for Cell Biology will benefit because they will be able to communicate this work to the wider scientific community and the public through their public engagement activities.

The EARLY CAREER RESEARCHERS using the facility will benefit from the high level imaging skills in both light and electron microscopy they will gain and multidisciplinary interactions with other users. These skills will transfer into their future careers in whatever sector they work. 'UK plc.' will benefit from such well-trained cross-disciplinary scientists who will be suited to many avenues of research.

INDUSTRIAL SCIENTISTS who are interested in how cellular processes are carried out and who are interested in the structure of nanoparticles, pharmaceuticals and agrochemicals will benefit from the new high resolution imaging facility.

Academics and early career researchers will continue their programme of SCIENCE COMMUNICATION WITH THE GENERAL PUBLIC, engaging with local schools, local and national media, science fairs, IGGY (International Gateway for Gifted Youth), MoleClues, open days etc. thus benefiting the level of education and of science debate outside academia as well as within. Encouraging scientists to take part in such activities early in their careers will establish habits and expertise for benefitting the general public that will stay with them for their careers. This impact will be immediate as well as over the next few decades.