Studying biology one molecule at a time by combining optical tweezers, fluorescence microscopy and microfluidics.

The purpose of our research proposal is to foster new and better ways to do science. We aim to empower scientists at the University of Cambridge and in the wider UK scientific community by giving them access to a new and important technology, that is transforming the way in which we design and carry out experiments.

Traditionally, molecular biologists have learnt about the way molecules such as proteins and DNA work inside cells by examining samples that contain a very large number of the molecules under study. This type of 'biochemical reconstitution' experiments have been instrumental in providing us with the molecular picture of the cellular world that we now have.

One important limitation of these experiments is that, because they consider a large population of the molecule under study, they can only tell us about its average properties over time. However, important aspects of molecular behaviour can only be understood when the molecules are considered individually. Important examples include how ATPase synthase, the enzyme that generates the energy-rich molecule ATP, works and how flagella propel bacteria forward, helping them swim in a purposeful way.

The instrument that we intend to acquire is akin to a set of tweezers to manipulate very tiny objects. To be able to handle something as small as a single molecule accurately and precisely, the tweezers are controlled by by laser light, hence the instrument name as 'optical tweezers'. By being able to grip and probe individual molecule with the tweezers, we can probe and understand their mechanical properties, which are important to determine the way molecules behave inside the cells.

The scientific applications of this new technology are very wide, and include understanding how proteins that protect our genome from damage interact with DNA, and how cells transport their protein cargoes from one organelle to another. In turn, such knowledge can be eventually exploited to understand what happens to us when such important cellular processes are disrupted in disease, and how to design rational strategies to intervene therapeutically.

Recent impressive technological developments have dramatically improved our ability to study biological processes at single-molecule level in a controlled and quantitative manner. Consequently, our understanding of the biochemical transactions underpinning cellular behaviour has been transformed by the single-molecule revolution that has taken place over recent years.

Until recently, single-molecule analysis was restricted to dedicated laboratories with the necessary expertise in engineering and biophysics to build and maintain the required instrumentation. This situation changed recently with the introduction by Lumicks Ltd of the C-trap, the first commercially available single-molecule optical-tweezers instrument. We propose to purchase a C-trapTM G2 instrument (https://lumicks.com/products/c-trap-optical-tweezers-fluorescence-label-free-microscopy/), which combines optical tweezers, fluorescent microscopy and microfluidics to image and manipulate individual molecules with a high degree of special and temporal resolution. The C-trap been designed to allow direct and routine access to single-molecule techniques to researchers in the Life Sciences.

The range of ongoing scientific projects in the wider Cambridge community that will benefit from the C-trap technology is very wide, and includes the study of protein-DNA/RNA interactions, protein engineering, cellular transport, bacterial biofilm structure and small-molecule therapeutics. A detailed description of the projects and experiments that will become possible through access to the C-trap is provided in the Case for Support.

Initially, the most significant effect expected from the availability of single-molecule optical-tweezers technology will be the profound academic impact that it will have on both both academic and industrial researchers in the University of Cambridge and local Cambridge area. We expect that the impact will become rapidly evident because the C-trap will enable excellent new research leading to scientific advances in radically different areas of Molecular Biology.

Adoption of this new technology will also make an important contribution towards maintaining the current position of excellence of the UK in the Life Science. No optical-tweezers system with the exquisite level of single-molecule manipulation afforded by the C-trap is available in Cambridge. Indeed, the only C-trap instrument in the UK was recently obtained by Imperial College. This contrasts with the situation worldwide, where several
such instruments are already available, most prominently in continental Europe and the US.

In the longer term, it is expected that these advances will have clear positive economic and societal impact, in terms of translating the novel findings into for instance potential therapies for patients and into new products and procedures of commercial value for the biotech industry. In this respect, we believe that it is highly significant that a number of local biotech companies have joined our application, as they see value in the adoption of C-trap technology in their basic research aimed at developing small-molecule compounds of therapeutic value.