An advanced multi-purpose instrument for biological ultrafast time resolved spectroscopy

Dynamical processes, including those on the ultrafast timescale, underpin much of the world around us. In biology they contribute to the white noise of vibrational motions inherent to large macromolecules and their environment, and also the 'coloured noise' that can assist or drive structural and biochemical change. Biologists normally consider biological processes on the timescale of seconds, minutes and hours, and examples include protein folding, biological catalysis, signalling, molecular and cellular motions. These processes, however, depend on the behaviour and interaction of electrons and the motions of the molecules/atoms in which they reside. Processes that occur on much faster timescales - typically picosecond to femtosecond timescales - dictate these properties. Crucially, understanding of biological processes requires mechanistic understanding of dynamical changes on the picosecond to femtosecond timescales i.e. with a time resolution of 1,000 million millionths of a second. This requires the development of specialised ultrafast laser spectrometers that can record chemical change, vibrational motion and the movement of electrons on these very short timescales.

The ultrafast timescale is where fundamental physics meets biology. Fundamental knowledge of chemical and physical properties on the ultrafast timescale underpins our understanding of ALL biological properties on the microsecond to second timescales (and beyond). Femtosecond lasers have been developed and utilised in the physical sciences and increasingly are having major impact in the biological sciences. Early wins include demonstration of 'quantum wierdness' (e.g. entanglement and coherence) in biological photosynthesis, and analysis of early chemical events in biological catalysis. These ultrafast methods provide versatile tools with wide-ranging applications. In this proposal we aim to establish a unique multi-functional ultrafast instrument, supported with expert physical and biological sciences expertise, that will transform a wide range of multi-user bioscience projects in the UK, by exploiting the power of ultrafast spectroscopy. This will provide crucial fundamental knowledge that will allow, for example, the design and exploitation of bioscience components in biotechnology and industrial applications, and contribute to the emerging science of synthetic biology through knowledge-based design.

We propose to establish a unique facility, by providing a multi-purpose ultrafast pump-probe instrument, with multiple detection modes and a wide range of applications in bioscience research. The instrument/facility will have both UV-visible and IR transient absorption and fluorescence up-conversion capabilities. We will establish a community-based advanced instrument, supported by expert experimental scientists for ultrafast time-resolved spectroscopy (UV-vis, IR and fluorescence) of multiple biological systems. The instrument will comprise a one-box femtosecond Ti:sapphire amplifier laser system (1 kHz, 100 fs pulses, >3.5 W, 800 nm), which with associated OPAs, will produce wavelengths ranging from 250 nm to 10 micrometer. Pre-existing visible transient absorption and fluorescence up-conversion instruments will be run with this laser. A new infra-red transient absorption spectrometer will allow probing of the 3 - 10 micrometer region where biological systems have many vibrational modes. This equipment will be truly unique to the UK bioscience community and will provide significant infrastructure for many research groups both at Manchester and other centres in the UK. It will be made available to BBSRC-funded researchers from other academic institutes as well as industrial scientists. The applicants are exceptionally well funded (mainly BBSRC, but also EPSRC and other grant agencies) and engaged in biophysical/biochemical analysis of a range of systems. This equipment proposal is structured around these strengths which include: fundamental studies of enzyme mechanisms and catalysis; applications of light activation in industrial biotransformations and biocatalysis; mechanisms and engineering of light-activated signaling proteins and applications as novel components in synthetic biology; mechanistic studies and applications of proteins at novel materials interfaces (e.g. graphene); studies of ultrafast dynamics underpinning component design for synthetic biology.

We will establish a multi-purpose ultrafast spectroscopy instrument to support a wide range of programmes in the UK bioscience community with a wide range of detection capabilities including UV-visible and IR, and fluorescence up-conversion spectroscopies. The bioscience community currently lacks a multi-user instrument managed by expert staff to support a wide range of bioscience projects. The proposed instrument/facility would therefore benefit a wide range of users (expert and non-expert) from the UK bioscience community who will have access to the power of ultrafast spectroscopy in a fully supported and multi-functional facility.

Therefore the research that will be enabled by the acquisition of this ultrafast spectroscopy facility will be far-reaching and would have potential benefit to: (i) new and existing academic researchers who are utilising ultrafast methods across the remits of BBSRC's committees; (ii) governmental and private sector scientists interested in design novel biological assemblies for applications in biotechnology and industry; (iii) private sector scientists from the time resolved spectroscopy (and software) industries, (iv) international organisations.

In order to allow the instrumentation to be fully exploited we would ensure that the equipment was fully accessible to all UK academics, who would also benefit from the high level of expertise and ultrafast specrtoscopy training that will be available at Manchester. Access to both internal and external users would be managed via a steering committee comprising at least three academic staff members (Scrutton, Hay, Munro), with Hardman and Heyes (who would provide hands-on help and training to the various projects), and an external partner member. Input would also be sought through establishment of a user group.

The impact of the ultrafast equipment/facility would be maximised by: (i) organising a project/facility specific workshops; (ii) generation of a web-portal for work conducted and opportunities for new investigators to the discipline; (iii) including details of the spectrometer on the N8 Shared Research Equipment Inventory to network with industry and via equipment.data.ac.uk; (iii) continuing our industrial collaborations; (iv) presentations to the scientific community via conferences and publications; (v) public engagement.

We will measure the success of our impact activities by the number of: (i) registered users of our community website/podcasts and associated outreach materials; (ii) applications to use the ultrafast facility; (iii) scientific publications and citations thereof relating to the facility; (iv) participants that attend our workshops and future collaborations with new external partners.