A Chemical Imaging Platform for Discovery Biosciences (CIP-DB).

Lead Research Organisation: University College London
Department Name: Cell and Developmental Biology

Abstract

Understanding the biology of cells, their component molecules and behaviors and characteristics when aggregated into tissues has always been a key goal in life science and medicine. Advances in microscopy are profoundly changing the way cells, sub-cellular structures, molecules and tissues can be studied. For a research intensive institution, such as UCL, to remain a competitive leader in biomedical sciences, it is essential to adopt transformative imaging technologies when their power is recognized, the technology is affordable, well understood and established as viable for transformative bioscience work. Furthering our understanding of health and diseases requires a deep knowledge of the cell, its constituents and their development from very early stages of life. How to image intact, large-scale, live or preserved postmortem cells, tissues and organisms at high spatial and spectroscopic resolution with and sensitivity remains a key challenge.

UCL's international reputation as a key contributor in bioscience/biomedical discoveries has, in part, been successful because of it's sustained commitment to be an early adopter of new, transformative technologies. One new and powerful pathway toward this goal is to image directly (without the use of reporter probe molecules) the chemical and biochemical properties of cells and tissues over time. This must be done with equipment that is capable of capturing at high speed images with sufficient fine spatial detail and at very high chemical resolution so that hitherto, unknown processes can be discovered and studied. Raman imaging technology is particularly suited to the study of key distibutions and properties of biomolecules during embryogenesis, tissue differentiation, changes in phenotype during health and disease and performance of their biochemical function.

CIP-DB will add a new dimension of support to cutting edge research in life and biomedical sciences. The Raman microscope is a unique system that enables acquisition of highly detailed, qualitative and quantitative information on the chemical composition of a tissue or cellular sample pixel-by-pixel. From this data images displaying the distribution of chemical species much like a multi-idimensional virtual staining, but without the introduction of probes, dyes, antibodies or genetically encoded markers, can be constructed. These images provide rich biochemical and molecular biological information and their spatial distribution. This characterization of biological samples is essentially orthogonal to that collected through other light microscopic techniques like the fluorescence imaging used almost universally and exclusively in modern cellular biosciences. Furthermore confocality and near infrared illumination will allow deep probing of samples and 3D chemical image reconstruction. Contour tracking technology will allow application of the technique to non-flat, rough or complex surfaces. This will allow unprocessed tissue samples to be analysed as well as samples where the thickness is changing over the duration of the experiment through growth, swelling or other changes.

The main co-applicants on this proposal are, therefore, research groups at UCL and colleagues in nearby research institutes whose focus is on aspects of molecular cell and developmental biology and biochemistry trying to elucidate the fundamental processes and key events/factors that underlie organismal development in both health and disease. In addition, the microscope will be accessible to a large number of diverse research projects addressing many outstanding questions in fundamental biosciences. The requested funds will enable us to obtain a one of the leading Raman microscope that will significantly enhance our research capabilities and lead to new scientific discoveries.

Technical Summary

Many researchers within the Faculty of Life Sciences require access to state-of-the-art imaging equipment for their studies. However, current core access across UCL lacks precision instrumentation for studying the chemical composition of cells and tissues in cultures/embryos/larvae/tissues/organs or during developmental processes. Modern, state of the art instruments have enhanced sensitivity and speed of acquisition, signal acquisition and analysis technologies. We will combine this with our advances in sample processing and mounting tecniques that greatly reduces the costs of sample presentation, enhances signals and optimizes processing of notoriously difficult samples of the type used predominantly in anatomy, embryology and anatomical pathology. Our new approaches also allow the use of higher laser power to accelerate scanning with maximum signal to noise ratio. Importantly the lower power, low phototoxicity at the near infrared laser wavelengths are most useful for Raman imaging of living cells, organisms, embryos, 3D cell cultures and organoids, and allow for long-term time-lapse imaging. The requested CIP-BD will also feature fluorescence imaging so that investigators can locate subcellular or tissue regions of interest flagged by antibodies, genetically encoded GFP or other beacons. All of these properties together will provide powerful advantage for understanding the relationships between the fundamental biochemistry of cells and its heterogeneity across tissues or stages of differentiation and development. Our research groups will have open, well technically supported and easy access to the equipment so that we can image across a wide biological spectrum - dictystellium, C.elegans, chicks, brittle stars, sea urchins, large animals and humans and organs and organiods.

Planned Impact

The assembled multidisciplinary team encompasses diverse fields and gathers together expert and experienced scientists each leading their own groups of researchers, networks, interest groups or departments that can make immediate use of the biochemical imaging platform proposed here. They will use a wide verity of model and human systems to investigate their particular research questions by capitalising on the unique capabilities of Raman microscopy. The areas of expertise of the groups are mainly in the field of biosciences therefore the potential Impact of these studies could be significant for a number of stake holders. However participants are also drawn from statistical and computational sciences and chemistry allowing the team to access know how and research power from engineering, computing, data and physical sciences - we will not be limited to the software and hardware tools supplied by the instrument manufacturer - we can develop new tools on our own. Furthermore we will be able to use the resources of these non-bioscience departments to access wider communities of expertise including interdisciplinary PhD students, postdocs and early career researchers interested in the new opportunities for analytical work thrown up by dedicating a high-end Raman microscope to projects in the extraordinarily rich biomedical research environment to be found at UCL and in the wider Bloomsbury area e.g. The Royal Veterinary College and Francis Crick Institute for impact on animal and human health and disease research respectively The instrumentation will facilitate a wealth of on-going research projects across the whole of UCL and catalyse the development of new initiatives once we spread the news of its power, accessibility and expert technical support through integration into UCL B-CALM. This will inevitably lead to more scientific discoveries with the consequential effects that these discoveries will impact on fundamental biosciences and studies of human and animal health and disease.

The outcomes of these projects will transfer knowledge and the tools generated using in vitro and vivo approaches to a wide scientific audience, both in academia and industry and may eventually impact on scientific discovery. Through a recent, successful InnovateUK award we obtained a prototype Raman microscope instrument from the leading UK Raman instrument manufacturer (Renishaw PLC) equipped with a single laser line and a single fixed 50x objective we have generated all of the data underpinning this application. With the higher power platform instrument described here we will extend our capability to examine living various living cells, embryos and other biological materials and at very much greater spatial, temporal and chemical resolution. Importantly because of our existing relationship with Renishaw we will feedback to this leading UK manufacturer real-world challenges raised by the range of bioscience experiments already planned or emerging in response to provision of the new platform. This exposure to research requirements and challenges will allow them to respond with engineering and technical innovations valuable to use, other researchers and innovators and future customers.

We expect that this research will also affect society as a whole in the long-term by influencing approaches of regenerative medicine, tissue engineering, diagnostics, therapeutics, and morphogenesis. It is important, however, that our impact activities also target academic audiences, as further research and application of our findings will be essential on the pathway to economic and societal impact. Moreover, the outcomes of the proposed research will not only support BBSRC Strategic Research Priorities but will also contribute to training and capacity building and hence amplify the impact of BBSRC enabling themes on the next generations of bioscience researchers.

Publications

10 25 50