13TSB_TIBio: Enabling Technologies for Advanced Multiphoton Microscopy

A major challenge in fundamental biological research and the development of biotechnology is to measure heterogeneity in chemical composition at the cellular and subcellular scale in living cells and tissues. Gaining the ability to visualise the distribution of compounds at this scale would represent a major advancement. The current standard in biological imaging, fluorescence microscopy, provides chemically specific image contrast by molecularly targeted probes. However, these probes are too bulky for labelling small molecules such as lipids, carbohydrates, metabolites and many drugs that play essential roles in the function of living cells and tissues. Coherent Raman scattering (CRS) microscopy has emerged as powerful tool to generate signals from vibrational spectroscopy to provide label-free mapping of biomolecules in real-time. However, due to the specialised laser requirements, this technique is currently confined to a handful of specialised laboratories with the resources and expertise to operate such laser systems. This project aims to develop a simple, compact module that will upgrade pulsed lasers currently used in biological microscopes to provide wider access to CRS imaging.

A major challenge in fundamental biological research and the development of biotechnology is to measure heterogeneity in chemical composition at the cellular and subcellular scale in living cells and tissues. Gaining the ability to visualise the distribution of compounds at this scale would represent a major advancement. The current standard in biological imaging, fluorescence microscopy, provides chemically specific image contrast by molecularly targeted probes. However, these probes are too bulky for labelling small molecules such as lipids, carbohydrates, metabolites and many drugs that play essential roles in the function of living cells and tissues. Coherent Raman scattering (CRS) microscopy has emerged as powerful tool to generate signals from vibrational spectroscopy to provide label-free mapping of biomolecules in real-time. However, due to the specialised laser requirements, this technique is currently confined to a handful of specialised laboratories with the resources and expertise to operate such laser systems. This project aims to develop a simple, compact module that will upgrade pulsed lasers currently used in biological microscopes to provide wider access to CRS imaging.

The proposed project has the potential to impact in the following areas:
1) Academic Research Community: The immediate beneficiaries of this laser module will be academic researchers who currently use multiphoton fluorescence microscopy. The module will provide a simple upgrade allowing the enhanced capability of coherent Raman scattering microscopy. Multiphoton microscopy is widely used across a broad range of biomedical research areas, all of which could benefit from having access to CRS microscopy. Specific areas where CRS has already been shown to offer advantages over current microscopies include; neuroscience, cancer research, fundamental cell biology, lipid biology, and drug delivery. In the longer-term, the widespread adoption of CRS microscopy amongst these research communities will facilitate the greater understanding of fundamental biology in these important areas.
2) Industrial Stakeholders: The same unique capabilities offered by CRS microscopy that are desirable for academic research also have applications in many areas of commercial R&D. Key industrial research and development sectors that have adopted multiphoton fluorescence microscopy and that would benefit from CRS microscopy include the pharmaceutical, agrochemical, consumer product, and cosmetic industries.
3) Students: Exposure to cutting-edge research is something that can really inspire school and undergraduate students to pursue a career in science. The proposed project will be an excellent demonstration of the value of innovative collaboration between academia and industry.

The following activities will take place to ensure that the project achieves its potential impact.
1) Impact Workshop on Coherent Raman Scattering Microscopy: To highlight the capabilities offered by CRS to both beneficiaries academic and industrial researchers we will hold a 1-day workshop at the University of Exeter on coherent Raman scattering microscopy. The workshop will include a keynote presentation from a leading overseas expert on CRS microscopy and invited presentations from UK and European CRS groups. The module will feature strongly in the workshop, being the focus of practical demonstrations of CRS microscopy in the Exeter laboratory. Attendees will be given the opportunity to bring samples on which to explore the feasibility of CRS microscopy. Funding to cover the travel expenses of the keynote speaker and light refreshments are requested for this activity.
2) Continued Engagement with User Community: Engaging with the biomedical user community beyond the duration of this project is vital for the development of future innovations in advanced optical microscopy. Exeter will continue to hold workshops on the application and development of CRS microscopy beyond the lifetime of this project. These will allow biological researchers who have adopted CRS to share there results and experiences those who are new to the technique. The PI has previously held similar events that were funded through commercial sponsorship.
3) Outreach: The University of Exeter actively encourage young people into science and regularly take secondary school students on work experience placement. This gives valuable experience and insight to teenagers considering science as a career and will help attract young people into scientific careers. In addition to this tours of the facility (including demonstrations) will be incorporated into the science summer schools for 16- 18 year olds that are currently run by the University. The interaction between M Square Lasers and the University of Exeter will be used to highlight how the interaction between academia and industry to exploit commercial applications of cutting-edge research techniques.