Multidisciplinary Super Resolution Microscopy Facility at Nottingham University
Lead Research Organisation:
University of Nottingham
Department Name: Sch of Molecular Medical Sciences
Abstract
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.
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.
Technical Summary
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.
Planned Impact
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.
-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.
Organisations
Publications
Petch JE
(2021)
Combining Inducible Lectin Expression and Magnetic Glyconanoparticles for the Selective Isolation of Bacteria from Mixed Populations.
in ACS applied materials & interfaces
Kilpatrick LE
(2017)
Real-time analysis of the binding of fluorescent VEGF165a to VEGFR2 in living cells: Effect of receptor tyrosine kinase inhibitors and fate of internalized agonist-receptor complexes.
in Biochemical pharmacology
Sherman HG
(2019)
Mechanistic insight into heterogeneity of trans-plasma membrane electron transport in cancer cell types.
in Biochimica et biophysica acta. Bioenergetics
Nizamudeen Z
(2018)
Rapid and accurate analysis of stem cell-derived extracellular vesicles with super resolution microscopy and live imaging.
in Biochimica et biophysica acta. Molecular cell research
Harrison RP
(2017)
Enhancing cell and gene therapy manufacture through the application of advanced fluorescent optical sensors (Review).
in Biointerphases
Roques M
(2019)
Plasmodium centrin PbCEN-4 localizes to the putative MTOC and is dispensable for malaria parasite proliferation.
in Biology open
De Melo N
(2019)
Live Simultaneous Monitoring of Mineral Deposition and Lipid Accumulation in Differentiating Stem Cells.
in Biomimetics (Basel, Switzerland)
Willson BJ
(2016)
Production of a functional cell wall-anchored minicellulosome by recombinant Clostridium acetobutylicum ATCC 824.
in Biotechnology for biofuels
Kilpatrick LE
(2019)
Complex Formation between VEGFR2 and the ß2-Adrenoceptor.
in Cell chemical biology
Jacot D
(2016)
An Apicomplexan Actin-Binding Protein Serves as a Connector and Lipid Sensor to Coordinate Motility and Invasion.
in Cell host & microbe
Samperi M
(2020)
Enhancing Singlet Oxygen Generation by Self-Assembly of a Porphyrin Entrapped in Supramolecular Fibers
in Cell Reports Physical Science
Nizamudeen Z
(2021)
Low-Power Sonication Can Alter Extracellular Vesicle Size and Properties
in Cells
Samperi M
(2019)
Quantification of energy of activation to supramolecular nanofibre formation reveals enthalpic and entropic effects and morphological consequence.
in Chemical science
Eleftheriou M
(2015)
5-Carboxylcytosine levels are elevated in human breast cancers and gliomas.
in Clinical epigenetics
Ramsawhook A
(2017)
Medulloblastoma and ependymoma cells display increased levels of 5-carboxylcytosine and elevated TET1 expression
in Clinical Epigenetics
Belsham HR
(2019)
Identification of key residues that regulate the interaction of kinesins with microtubule ends.
in Cytoskeleton (Hoboken, N.J.)
Romero M
(2022)
Fluid dynamics and cell-bound Psl polysaccharide allows microplastic capture, aggregation and subsequent sedimentation by Pseudomonas aeruginosa in water.
in Environmental microbiology
Peach CJ
(2018)
Molecular Pharmacology of VEGF-A Isoforms: Binding and Signalling at VEGFR2.
in International journal of molecular sciences
Al-Natour MA
(2019)
LC-MS metabolomics comparisons of cancer cell and macrophage responses to methotrexate and polymer-encapsulated methotrexate.
in International journal of pharmaceutics: X
Canales Coutiño B
(2020)
A Genetic Analysis of Tumor Progression in Drosophila Identifies the Cohesin Complex as a Suppressor of Individual and Collective Cell Invasion.
in iScience
Zeeshan M
(2020)
Real-time dynamics of Plasmodium NDC80 reveals unusual modes of chromosome segregation during parasite proliferation.
in Journal of cell science
Dunajová L
(2016)
The NH2 terminus of survivin is a mitochondrial targeting sequence and C-Src regulator
in Journal of Cell Science
Lugo Leija HA
(2020)
Cold-induced beigeing of stem cell-derived adipocytes is not fully reversible after return to normothermia.
in Journal of cellular and molecular medicine
Faidra Angelerou MG
(2020)
Mechanistic investigations into the encapsulation and release of small molecules and proteins from a supramolecular nucleoside gel in vitro and in vivo.
in Journal of controlled release : official journal of the Controlled Release Society
Ramsawhook AH
(2017)
Immunostaining for DNA Modifications: Computational Analysis of Confocal Images.
in Journal of visualized experiments : JoVE
Zeeshan M
(2022)
Plasmodium SAS4: basal body component of male cell which is dispensable for parasite transmission.
in Life science alliance
Soave M
(2020)
Application of Fluorescent Purinoceptor Antagonists for Bioluminescence Resonance Energy Transfer Assays and Fluorescent Microscopy.
in Methods in molecular biology (Clifton, N.J.)
Abakir A
(2021)
Detection of Low-Abundance DNA Modifications Using Signal Amplification-Based Immunocytochemistry.
in Methods in molecular biology (Clifton, N.J.)
Rajani S
(2021)
Computational Analysis of DNA Modifications in Confocal Images.
in Methods in molecular biology (Clifton, N.J.)
Johnson L
(2019)
Low Molecular Weight Nucleoside Gelators: A Platform for Protein Aggregation Inhibition.
in Molecular pharmaceutics
Martinez De La Cruz B
(2021)
Modifying the m6A brain methylome by ALKBH5-mediated demethylation: a new contender for synaptic tagging.
in Molecular psychiatry
Samperi M
(2021)
Light-controlled micron-scale molecular motion.
in Nature chemistry
Zeeshan M
(2023)
Plasmodium ARK2 and EB1 drive unconventional spindle dynamics, during chromosome segregation in sexual transmission stages
in Nature Communications
Abakir A
(2020)
N6-methyladenosine regulates the stability of RNA:DNA hybrids in human cells.
in Nature genetics
Abakir A
(2020)
N6-methyladenosine regulates the stability of RNA:DNA hybrids in human cells.
in Nature genetics
Martinez De La Cruz B
(2023)
m6 A mRNA methylation in human brain is disrupted in Lewy body disorders.
in Neuropathology and applied neurobiology
Self T
(2015)
Super resolution microscopy - the bright side of molecular biology
in Pharmacology Matters
Harrison RP
(2019)
Intracellular processing of silica-coated superparamagnetic iron nanoparticles in human mesenchymal stem cells.
in RSC advances
Wojciechowska M
(2018)
Quantitative Methods to Monitor RNA Biomarkers in Myotonic Dystrophy.
in Scientific reports
Velickovic K
(2019)
Caffeine exposure induces browning features in adipose tissue in vitro and in vivo
in Scientific Reports
Velickovic K
(2018)
Low temperature exposure induces browning of bone marrow stem cell derived adipocytes in vitro.
in Scientific reports
| Title | Biodiscovery Institute Image Competition |
| Description | We have used the samples form research (bacteria, scaffolds, bio materials, cancer cells) and non- research, such as Dandelion, or duckweed, and naturally occurring bio films to create artistic images. |
| Type Of Art | Artistic/Creative Exhibition |
| Year Produced | 2020 |
| Impact | We have created artistic material (images of high visual impact) for the Launch event of the Biodiscovery institute, organised an image competition. Images from supper resolution microscope won the competition, as well as more shortlisted images form the images will be used in the exhibition. This will go public in the Highfields park on large outdoor prints. |
| Title | Competition |
| Description | We have launched an image competition where users of the microscopes submit their best work to the imaging team. Selected images are printed on mugs and prints and displayed on the walls too. |
| Type Of Art | Image |
| Year Produced | 2018 |
| Impact | Images attract new users and advertise microscopy as a tool for research. |
| URL | https://www.nottingham.ac.uk/life-sciences/facilities/slim/slim-image-gallery.aspx |
| Title | Duckweed root decay - Nikon Small World Image of Distinction |
| Description | Duckweed root decay (DNA shown in blue-cyan and chlorophyll pseudo colored to red) Dr. Robert Markus We used super resolution microscopy to show a naturally developing biofilm on a plant material entering decomposition. |
| Type Of Art | Image |
| Year Produced | 2020 |
| Impact | Viewers are engaged and we show an example of the potential of the super resolution microscope we use on daily basis fro research. |
| URL | https://www.nikonsmallworld.com/galleries/2020-photomicrography-competition/duckweed-root-decay-dna-... |
| Title | Hebe plant anther with pollen - Nikon Small World Winner Entry |
| Description | 2020 Photomicrography Competition Hebe plant anther with pollen Dr. Robert Markus Zsuzsa Markus We have created artistic view of an object *anther of a flower- to make awareness of the natural world around us which can be found in the university campus. |
| Type Of Art | Image |
| Year Produced | 2020 |
| Impact | The viewers are made aware of the potential of the uper resolution microscopy unit funded by this award. |
| URL | https://www.nikonsmallworld.com/galleries/2020-photomicrography-competition/hebe-plant-anther-with-p... |
| Title | High resolution prints of super resolution microscopy images |
| Description | A selection of super resolution images have been printed and mounted on the corridors of the University. The images were captured using the instrument (Elyra PS.1 super resolution microscope) acquired using the founds of this grant. |
| Type Of Art | Image |
| Year Produced | 2015 |
| Impact | Each image has a short description, explaining the biology and the super resolution technique applied. |
| Title | Launch event of Biodiscovery Institute |
| Description | Images and scientific story were collated and printed on large boards at University Highfields Park as an outdoor exhibition, featuring images produced at the super resolution microscope facility. |
| Type Of Art | Artistic/Creative Exhibition |
| Year Produced | 2021 |
| Impact | The general public could see the images and the science we run and made aware of the various research topics, enhanced by the impactfull images. |
| Title | Super resolution images printed and displayed |
| Description | The winning entry of by Robert Markus was printed and displayed permanently in the building hosting the microscope. |
| Type Of Art | Image |
| Year Produced | 2018 |
| Impact | The image has an excellent visual impact, therefore fellow researchers ask about it, how it was achieved and where, thus we have another chance to advertise our super resolution microscope and propose them to use and do research with this high end instrument. |
| Description | We further improved the super resolution imaging platform by • Screening and selecting new chemicals for sample preparation (ms. in prep) • Implemented new data analysis techniques • Trained more scientists to use the instrument independently • Teaching has been improved by making more teaching tools (slides, video tutorials, internal use at the moment, instrument specific) • Started new imaging projects and initiated new collaborations including with Industrial partners • We regularly participate at the outreach events Scientific Projects (selected examples) Overall in the last 7 years we had couple of hundred users with multiple imaging projects on the multidisciplinary Super Resolution Microscope. 1. Discovered new molecules involved in the life cycle (division) of the malaria parasite in a research which aims to identify molecular targets combat infections 2. Developed a method to visualise and analyse very small (nanometre sized) vesicles, which are shed from cells. These contain molecular messages governing neuronal development (out publication being cited by Cell and others, and method became as a benchmark in EV biology). 3. Several projects working in molecular pharmacology identified and visualised nano-scale structures used in drug delivery systems, these have been characterised with super resolution microscopy too. 4. Continued working on RNA biology in relation to genetically inherited diseases 5. Continued the work on biofilm models including human tissue 6. Screened for chemicals to be used in sample preparation in order to achieve the best possible resolution results are implemented in the projects, a manuscript is in preparation on this 7. Continued the work on localisation of nano-particles in cells and investigated drug delivery system in vitro. 8. Developed a new nano-calibration test slide sample for super resolution microscopy. The prototypes are ready and we are working on IP protection and accelerate commercialisation. This sample is aimed to serve as a standard to compare various super resolution microscopy systems. 9. We got new molecular imaging insight (single molecule imaging) into murine spinal cord and brain; the research aims to understand pain at the highest resolution possible using our multi tool microscope and different imaging modalities. Results are used to attract further funding. 10. We advised and worked with commercial partners to investigate, visualise, and demonstrate how their product works. For example development of gloves with antimicrobial activity. We performed live experiments at the worldwide launch event. 11. Design tracer molecules which bind to the surface of the human pathogen Staphylococcus aureus. Super resolution microscopy will help to investigate these proteins at 10 times more resolution than before enabling us to gain a better understanding of the infections caused by this bacterium 12. Localised proteins in E. coli involved in disease. We have constructed a three dimensional super resolution model which has enabled us to measure the mobility these proteins inside the bacterial cell using super resolution microscopy alongside in situ ?uorescent labelling of individual molecules. 13. Generated high resolution three dimensional models to study the distribution of lipids in the microalgae (C. reinhardtii) used as functional food and feed. 14. Localised nanoparticles after treatment. These experiments show the efficiency of various drug delivery strategies. With super resolution microscopy were able to visualise and measure fluorescent nanoparticles inside different cell lineages. 15. Using this instrument (TIRF + SuperRes) made possible to discover and characterise photo-responsive nanoutubes 16. We are pioneering live imaging and super resolution imaging of biofilms on tissue models 17. Working with commercial partners to characterize genetically engendered bacteria to be used in industry 18. The technical knowledge Robert Markus and the imaging team gained by running this instrument on daily basis is invaluable, and it is disseminated an used withing the University and UK imaging community. 19. Neurology related: molecular imaging of structures (synapses) within in the the mouse spinal chord have been investigated, results published, and investigation is continued by members of the Arthritis UK Pain Centre. |
| Exploitation Route | Our findings will be applicable to researchers and applications in the following areas: protein localisation, antimicrobial biology, novel antibiotics and bio-materials, drug delivery, lipid localisation, biofilm biology, regenerative medicine, nano materials and more general in super resolution microscopy. |
| Sectors | Agriculture, Food and Drink,Chemicals,Digital/Communication/Information Technologies (including Software),Education,Environment,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Description | Researchers in the Midlands now are aware of this instrument, and the output data and images have been seen at various forums, seminars, conferences, posters, workshops and publications. Projects, funding requests are submitted with the awareness and clear plans to use the instrument at full capacity. Having full time access, and dedicated person to run the super res facility, Robert Markus and Tim Self developed expertise and advise researchers, help in planning the projects, design experimental work, and help in sample preparation. All these are crucial steps to perform high quality publishable results. Leading researchers, (PI's) are aware now what the instrument and the imaging team is capable of, they regularly inquire for novel ideas, and send their PhD students to run pilot experiments then design a whole study using the instrument. Recently (2019-2020) a new building Biodiscovery Institute 3 (BDI) has been opened, and the Biodiscovery 1&2 and 3 is a growing community where scientific ideas are shared, a new research community is forming. We have been proactively reaching out to all the newly recruited research fellows and showcasing the microscope facility and / or already run experiments. The BDI had the opening and showcase ceremony last year, for which we prepared large prints and stories of various imaging projects and showcased the images in the Highfields Park University campus to the general public. Outstanding and internationally awarded images produced using this instrument were shown. |
| First Year Of Impact | 2016 |
| Sector | Agriculture, Food and Drink,Chemicals,Digital/Communication/Information Technologies (including Software),Education,Environment,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Cultural |
| Description | Development of Nano calibration sample for super-resolution microscopy (UNICAS programme) |
| Amount | £5,000 (GBP) |
| Organisation | University of Nottingham |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 04/2015 |
| End | 09/2015 |
| Description | National Biofilms Innovation Centre |
| Amount | £12,801,513 (GBP) |
| Funding ID | BB/R012415/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 12/2017 |
| End | 11/2022 |
| Title | Screening of embedding materials for super resolution microscopy |
| Description | We screened 96 different conditions, with 3 different sample types together with so called mounting media, in order to improve longevity, contrast and resolution of samples. We are preparing a research publication on this topic. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2019 |
| Provided To Others? | No |
| Impact | The outcome is already implemented in our institute, researchers are aware of the developments and they use the results to improve their samples. |
| Title | Super Resolution Microscopy Test Slide |
| Description | We have developed a test slide which can be used to assess, test, the performance of the Super resolution microscope. This tool will enable any user to test, and evaluate the workflow, the image analysis, and the instrument itself. As well as it is used for teaching and training purposes too. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2019 |
| Provided To Others? | No |
| Impact | We are taking fuhrer the prototype to became a product available to all microscopy facilities. |
| Title | Nanoscale calibration samples for super-resolution microscopy & Nanoscale photoactivatable particles for localisation and tracking of molecules of interest |
| Description | A new type of nanoscale calibration sample for super-resolution microscopy has been developed, that enables photo-activation and photo-switching via defined lsers. Detection of single molecule signals enable localisation and image reconstruction of nanoscale fluorescent objects of 100 nm. The nanoscale features, nanobeads, are modified with photoswitchable dyes, like spiropyran. They can be resolved via photo-activated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM) techniques in super-resolution microscopy due to their optical properties. The device can be easily manufactured at low cost and re-used over time for more than one experiment. The entire worldwide microscopy community using super-resolution microscopy could use such a sample in their laboratory to improve the calibration of their microscopes. The prototype will be finailsed in 2017. An Intellectual Property Review Form has been filed with the University of Nottingham. |
| IP Reference | |
| Protection | Protection not required |
| Year Protection Granted | 2016 |
| Licensed | No |
| Impact | At first glance the companies that could be interested in such technology would be companies that develop and work on the generation of new super resolution microscopes like Zeiss, Leica, Olympus or Nikon. Authors already have a contact in Zeiss that would be interested in testing the new samples. Beside, every laboratory in academia having a super resolution microscope could be interested in testing \ having such a sample.Authors have contacts over the country, research officers and academics, who would be happy to test the sample. |
| Title | Implementation of image data base and sharing platform |
| Description | We have implemented an image storage and sharing interface, where microscope users store their imaging data. The interface is using the Open microscopy environment (OMERO). Researchers use their university credentials to access, analyze, download or share their data. A backup system is also available, with a copy of all data. Access to this computer is restricted, permission can be obtained via the imaging team of School of Life Sciences, University of Nottingham. |
| Type Of Technology | Software |
| Year Produced | 2015 |
| Impact | Initially this image server has been implemented for the super resolution microscope, but in long term it will serve as a cross-disciplinary imaging platform. |
| URL | https://omero.nottingham.ac.uk/ |
| Description | 1st Elyra Super Resolution Microscopy Workshop London |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Since we have the Elyra super resolution microscope we have initiated a meeting between managers of similar microscope and the experts from the manufacturer (Zeiss). Experts of Super resolution microscopy from UK and Germany (Zeiss), met for the first Elyra Super Resolution meeting. The purpose of the meeting was to share our experience and difficulties and technical know-how we accumulated on the Super Resolution Microscope, which was acquired within this award. The attendees all work with similar or older instrument. Crucial technical details were shared, ideas exchange was very useful, additionally the representative of the manufacturer answered the questions of the audience. Robert Markus, gave a talk on his latest developments on refractive index of the mounting media of samples. |
| Year(s) Of Engagement Activity | 2017 |
| Description | 2nd Elyra Super Resolution Microscopy Workshop Norwich |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Imaging facility managers having similar microscope we have in Nottingham, met and discussed microscopy related topics. Representative of the manufacturer Zeiss also attended, and answered our questions related to the system. Novel super resolution microscopy technology was presented and debated. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Creating the image - filtering STORM data. Talk given by Robert Markus, on the implemenation of single molecule imaging (STORM). A case study and the software and analysis approaches were presented. |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Professional Practitioners |
| Results and Impact | Around 70 people attended, the talk. We had a froutful discussion on the development of the super resolution techniques, uset in our multidisciplinary imaging facilities. I ve got valuable input in further developint the projects, namely in the recorting frequency of the single molecules. Additionally we have discussed the technical details of our imaging system, I've engaged new collaborations on image analysis, and we will implemet the clustering algorithm in our projects. The audiance apprciated the techniques ive used to detirmine the negative control, and the techniques to eliminate the false positive hits, on our datatsets. |
| Year(s) Of Engagement Activity | 2017 |
| URL | https://sheffieldstorm.wordpress.com/ |
| Description | Frontiers if BioImaging |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | We attended the Frontiers if BioImaging meeting in Glasgow, where we met the experts in the filed of Super resolution microscopy and image analysis. Talks and one to one discussions expanded our knowledge, and we implemented the know-how on our return to Nottingham. |
| Year(s) Of Engagement Activity | 2018 |
| URL | https://www.rms.org.uk/discover-engage/event-calendar/frontiers-in-bioimaging-2018.html |
| Description | Frontiers of BioImaging |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | "Frontiers in BioImaging 2016 focused on the latest developments in applications of optical microscopy and image analysis across a range of biomedical fields. Sessions covered both technical developments and applications of these microscopy-based approaches to key cell and molecular biology questions. The meeting covered the key challenges in microscopy today: super-resolution imaging, phototoxicity and light sheet based methods, detection of in situ protein interactions and new tools for fluorescence visualisation and analysis." We met the leading experts of the field. Robert Markus learned about new computational methods for super resolution microscopy, the method was implemented in the facility. As well as interpenetration of super resolution results were discussed, and used in our publications regarding single molecule imaging. |
| Year(s) Of Engagement Activity | 2016 |
| URL | https://www.rms.org.uk/discover-engage/event-calendar/frontiers-in-bioimaging.html |
| Description | INTERNATIONAL SIMPOSIUM ON " GENE REGULATION IN BACTERIA WITH MEDICAL AND INDUSTRIAL INTEREST' Invited international speaker |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | The title of the invited talk was: 'Exploiting the therapeutic potential of targeting quorum sensing in Pseudomonas aeruginosa'. The symposium was organised by the Universidad Nacional Autonoma de Mexico in Mexico City. It was attended by master students, PhD students and research group leaders. |
| Year(s) Of Engagement Activity | 2018 |
| Description | International Seminar at the Universidad National Autonoma Mexico |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | This was a seminar to promote the results from the SENBIOTAR programme and identify new collaborations from the results of these programme as well as promote NBIC. The title of the seminar was: Versatility and targeting of quorum sensing signalling mechanisms There was a mixed audience of scientists |
| Year(s) Of Engagement Activity | 2018 |
| Description | Online Teaching |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | Using the knowledge generated over the years we designed and deliver microscopy related talks, where student learn about the principles of optical microscopy, the resolution limit and how Super Resolution microscopy works and practical examples are shown and discussed with examples from the ongoing research. |
| Year(s) Of Engagement Activity | 2020 |
| Description | Poster at 12th International Conference on Renewable Resources and Biorefineries, taking place in Ghent, Belgium on May 30-31 & June 1, 2016. |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | A Poster presentation has been given with the following title: Evaluation of Microalgae (Chlamydomas reinhardtii) for Human Consumption Patchaniya Eakpetch, Darwish Randa, Esra Kaya, Robert Markus, David Gray Images cerated using the supre resolution microscope had been presented. |
| Year(s) Of Engagement Activity | 2016 |
| URL | http://www.rrbconference.com |
| Description | Poster at Neuroscience @ Nottingham, 2016 |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | A poster had been presenetd with the following title: ""Immunofluorescent Imaging of RNP Markers in differentiated Neuronal Cell Lines using Super Resolution Microscopy" Braulio Martinez, Chris Gell, Robert Markus, Ian Macdonald, and Helen Miranda Knight Images created at the super resolution microscope were included. |
| Year(s) Of Engagement Activity | 2016 |
| URL | http://www.nottingham.ac.uk/neuroscience/index.aspx |
| Description | Poster at Sheffield STORM Symposium |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Professional Practitioners |
| Results and Impact | A poster competition has been taking place at the symposium. We have presented our multi colour STORM data. "Localization of Proteins of interest in E coli bacteria using two colour STORM" Robert Markus, Tim Self & Jafar Mahdavi |
| Year(s) Of Engagement Activity | 2017 |
| URL | https://sheffieldstorm.wordpress.com/ |
| Description | Poster at Young Life Scientists' Symposium 2016 - Advanced Light Microscopy |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Postgraduate students |
| Results and Impact | A poster with the following titles had been presented "Immunofluorescent Imaging of differentiated Neuronal Cell Lines using Wide-Field, Confocal, and Super Resolution Microscopy" Braulio Martinez, Chris Gell, Robert Markus, and Helen Miranda Knight |
| Year(s) Of Engagement Activity | 2016 |
| URL | https://www.bps.ac.uk/news-events/future-scientific-meetings/2016/young-life-scientists-symposium-20... |
| Description | Refractive index of the sample - get the best out of your microscpe |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Robert Markus, gave a talk on his latest developments on refractive index of the mounting media of samples. This is crucial to obtain the highest error free super resolution microscopy images and accurate data. Audience had useful comments and they had professional and very useful comments. Overall the ideas and the data met positive response therefore more work is invested to write up a manuscript with the aim to publish the technical results which were obtained at our super resolution microscope. |
| Year(s) Of Engagement Activity | 2017 |
| Description | Research Spotlight on the JPIAMR webpage |
| Form Of Engagement Activity | A magazine, newsletter or online publication |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other audiences |
| Results and Impact | The highlights from the SENBIOTAR programme were disseminated through a research spotlight in the JPIAMR website which is openly accessible to everybody. It highlights the success of this particular programme as a way of promoting the outputs from the JPIAM funding. |
| Year(s) Of Engagement Activity | 2019 |
| URL | https://www.jpiamr.eu/wp-content/uploads/2019/10/JPIAMR_Folder_3xCase_Screen.pdf |
| Description | Talk preseneted at The Microbiology Society Annual Conference 2016, 21-24 March at the Arena and Convention Centre (ACC), Liverpool, UK. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Result obtained using the super resolution microcope were presented in the Prokaryotic Cell Biology Forum on the afternoon of March 23rd in Liverpool at the Microbiology Society annual conference. "Localizing Autotransporter EspC during secretion to identify potential targets for novel antimicrobials" Mahmoud Ashawesh, Robert Markus, Christopher Penfold, C. and Kim R. Hardie. |
| Year(s) Of Engagement Activity | 2016 |
| URL | http://www.microbiologysociety.org/events/annual-conferences/index.cfm/annual-conference-2016 |