Award details

Multidisciplinary Super Resolution Microscopy Facility at Nottingham University

Reference	BB/L013827/1
Principal Investigator / Supervisor	Professor Miguel Camara
Co-Investigators / Co-Supervisors	Professor Cameron Alexander, Professor Jonathan Aylott, Professor Paul Barrow, Professor Malcolm Bennett, Dr Lee Buttery, Professor Chris Denning, Dr Philip Hill, Professor Stephen Hill, Professor Nigel Minton, Professor Clive Roberts, Professor Felicity Rose, Professor Kevin Shakesheff, Professor Renee Sockett, Professor Paul Williams, Professor Philip Williams, Professor Zoe Wilson, Dr Klaus Winzer, Professor Lorraine Young, Dr Ying Zhang
Institution	University of Nottingham
Department	Sch of Molecular Medical Sciences
Funding type	Research
Value (£)	735,267
Status	Completed
Type	Research Grant
Start date	18/12/2013
End date	17/05/2014
Duration	5 months

Abstract

The objective of this grant application is to provide the University of Nottingham with a Super Resolution Microscopy Unit in a Class 2 containment facility which can be used for both Class 1 and Class 2 experiments. The SR microscope will combine super resolution structured illumination microscopy (SR-SIM) and single molecule localisation techniques (PALM & dSTORM) to enable the localization of single molecules with precision down to 20 nm. Using this technology, an optical grid leading to a large number of images (e.g. 10,000) with very low levels of blinking fluorescence can be collected from single molecules by localising the dots in each of the 10,000 images. Because of the large number of acquired images required for the localisation data, samples in general need to be fixed unless lower localization precision is required. It will also provide particle tracking to follow single molecules in a sample without compromising resolution. Amongst the applications will be the ability to track single molecules (proteins, DNA, drugs etc). This microscope will be combined with a sophisticated confocal microscope with a 32-channel GaAsP detector for better quantum efficiency to improve the detection of weak signals and signal-to-noise ratio and which also enables the observation of molecules at high resolution in real time and the potential to image the same cell with SRM after fixation. This combination is essential for the acquisition of the molecular details of e.g. protein transport mechanisms, bacterial and viral proteins interacting with eukaryotic molecules, localization of small molecules in plant cells, biofilm architecture, tracking molecules in stem cell biology, DNA oregami structures, bacterial communication within biofilms, the activation of multiple nanosensors in specific areas of a bacterial biofilm and in tissue scaffolds etc.

Summary

For many years there has been a gap in the resolution of Confocal Laser Scanning Microsopy and even with perfect lenses, optimal alignment, and large numerical apertures the optical resolution of this form of light microscopy has been limited to approximately half of the wavelength of the light used meaning that only cellular structures and objects at least 200 to 350 nm apart could be resolved as distinct separate structures. However, much of the fundamental biology of the cell, occurs at the level of micro-molecular complexes in the size range of tens to few hundred nm, i.e., beyond the reach of conventional light microscopy. Super resolution microscopes (SRM) have been developed to break or bypass the classical diffraction limits and shift the optical resolution down to macro-molecular or even molecular levels. It can locate single molecules down to 20nm precision. SRM can also provide particle tracking to follow single molecules, e.g. proteins, DNA, drugs, in a sample without compromising resolution. Although the University of Nottingham (UoN) houses a range of microscopy facilities there is a clear gap in resolution that requires the cutting edge technology SRM provides to take the research into areas that are not currently achievable in Nottingham Life Sciences at present. Some of these areas include: (i) Microbiology: to understand better how bacterial cells interact with each other and how they can transfer molecules and signals within and between cells in bacterial communities known as biofilms; (ii) Bioenergy: to understand how some bacterial cellular energy reservoirs are transformed into biofuels and hence be able to optimize of these processes; (iii) Synthetic Biology: to understand how artificial cells and specialized DNA structures are developed as this can have a significant impact within different industrial contexts; (iv) Stem cell biology and tissue engineering: the SRM will have a significant impact on understanding the development of specialized cells from stem cells and the development of tissue scaffolds which will be paramount in regenerative medicine required for the repair of different damaged human tissues; (v) Plant sciences: to understand some of the signaling processes in root development and the mechanisms of pollen development which impact on crop production and sustainable agriculture; (vi) Food security: to unravel essential mechanisms of interaction between pathogens and host cells of farm animals which are responsible for significant loses in the farm industry with the ultimate aim of targeting some of these to reduce the negative economic impact they have and (vii) Molecular pharmacology: to facilitate the understanding of drug-target interactions enabling their optimization with the consequent improvements to health. Hence the acquisition of a SRM facility will enable UoN to address key biological questions in the above areas which will have a significant impact on science, the economy and society.

Impact Summary

The RCUK have identified an urgent need to invest in imaging technology through the RCUK Strategic Framework for Capital Investment. The establishment of next generation microscopy platforms are key to answering questions involving the multi-disciplinary research that underpins biology that utilises cutting-edge light microscopy and dynamic cell imaging. In this context the Super Resolution Microscope (SRM) facility will provide a unique dimension to the microscopy facilities at the University of Nottingham (UoN). Due to the multidisciplinarity of the research areas for which the SRM unit will be used, the main objective for this facility will be to impact on research, researchers and industry from the following disciplines: -Microbiology: the research proposed will have a significant impact in a wide range of industrial sectors where biofilms have either positive or detrimental effects. -Bioenergy Industry: it will enable a better understanding of how cellulosomes are transformed into biofuels resulting in the optimization of these processes and how to reduce product toxicity. -Synthetic Biology: the applications proposed will have an impact on the development of artificial cells and DNA nanostructures relevant to different industrial contexts. -Stem cell biology and tissue engineering: the SRM will have a significant impact on understanding the development of specialized cells from stem cells and the development of tissue scaffolds which will be paramount in regenerative medicine -Plant sciences: the proposed research will be important to understand signaling processes in root development and the mechanisms of pollen development which impact on crop production and sustainable agriculture. -Food security: the SRM will be used to unravel essential mechanisms of interaction between pathogens and host cells from farm animals which are responsible for significant losses in the farm industry with the ultimate aim of targeting these and reduce their negative economic impact. -Molecular pharmacology: the use of SRM will facilitate the understanding of drug-target interactions enabling their optimization with the subsequent impact on patients suffering so contributing to health. The SRM facility will also have an impact on: a. Researchers: Through the acquisition of formalized training on the use of the SRM facility. These will include not only researchers at UoN but also visitors from Nottingham collaborators needing to use SRM. b. The University of Nottingham: It will support the advancement of current research areas and the development of new research possibilities. Having a SRM will attract new collaborations to Nottingham, especially in areas where the current microscopy facilities do not provide the appropriate level of resolution c. The Midlands: as it will enable researchers from this region, especially those collaborating with the UoN to have access to a local SRM facility and associated expertise. d. The international relationships between UoN and researchers outside the UK through collaborations between Nottingham researchers and the international research community and industrial companies. The wider public will also benefit in the longer term from the research conducted at the SRM facility through the increased ability of different industrial sectors to respond to their customer needs from the environment, to agriculture and health. The research achievements from the SRM facility will be communicated to a range of audiences via presentations through to discussions and workshops with industry contacts, publications in journals targeting a wide range of audiences and conferences and a dedicated website. The research from the SRM facility and its potential will also be communicated to the general public through the yearly University of Nottingham May Fest and through the 'Nottingham Potential' outreach activities.

Committee	Research Committee B (Plants, microbes, food & sustainability)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	Advanced Life Sciences Research Technology Initiative (ALERT) [2013-2014]
Funding Scheme	X – not Funded via a specific Funding Scheme

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