Electron dynamics in metals and nanostructures take place on attosecond timescales. Until today, these extremely fast processes are little understood let alone utilized. With NearFieldAtto, strong-field driven phenomena at nanoscale metal structures will be explored to elucidate collective electron dynamics and to induce optical-field-driven currents -- on attosecond timescales. We will investigate the near-field of a nanotip, resulting from the collective dynamics, both in amplitude and phase. Conversely, we will use the tip as a nanometric sensor to map out the electric field inside the focus of a pulsed laser beam and will directly measure the local phase. In two-tip and molecular junctions, we will explore the ultrafast steering of electronic currents by optical fields, both over a nanometric gap and inside a molecule, taking advantage of the large near-field enhancement the systems offer. My group has recently shown that attosecond physics phenomena can be observed at solids, namely at nanoscale tips [Krüger et al., Nature 2011]. Hence, in NearFieldAtto we will employ techniques well known from attosecond physics with isolated objects, like gas-phase atoms and molecules, to steer laser-emitted electrons with the electric field of few-cycle laser pulses. We will use these electrons as nanometric probes to investigate optical properties of the solid state system and compare the results with those of isolated objects in gas-phase measurements. With two tips facing each other, we will realize a nanometric junction over which we will steer electrons with the optical field. A molecule placed between two tips will enable the investigation of a novel, ultrafast switching mechanism. NearFieldAtto will bring attosecond physics a leap forward as compared to the state-of-the-art, will introduce strong-field physics into (quantum-)plasmonics, and will open the door towards lightwave or petahertz nano-electronics in metallic and molecular nano-systems.

The goal of the proposed project is to employ novel states of matter for the development of new devices for quantum technologies, optoelectronics and ultrafast nanoelectronics. Speeding up and miniaturization of the existing electronics are approaching their physical limits. Novel states of mater are a rapidly growing field of science including quantum condensates and superconductors. One of the topics in the proposed project explores devices based on condensates of exciton-polaritons in semiconductors, representing both: ultrafast low-dissipation electronics due to their light-matter superfluid properties, as well as extremely nonlinear optics useful for quantum photonics. Another approach in the proposed project, which can provide a new direction in optoelectronic devices, is based on combining superconductors with semiconductors. The project takes advantage of the recent progress in high-temperature superconductors, which makes these technologies significantly more practical. Lately, a novel paradigm for finding new properties in the solid state has emerged - through the sudden change in topological invariants rather than breaking of symmetries. These topological phases of matter have been demonstrated to exist at the surface of some materials with strong spin-orbit coupling, revealing novel physical properties, including dissipationless spin currents with potential applications in spintronics and quantum technologies. A part of the proposed research focuses on devices based on proximity-induced high-temperature superconductivity in such topological insulators. This proximity effect has been predicted recently to produce the elusive Majorana fermion, which is of great interest for condensed-matter physics and quantum computation.

To address the challenges of solar energy capture and storage in the form of a chemical fuel, we will develop a hybrid photoelectrochemical-photovoltaic (PEC-PV) tandem device for light-driven water splitting. This concept is based on a visible light-absorbing metal oxide photoelectrode, which is immersed in water and placed in front of a smaller-bandgap thin film PV cell. This tandem approach ensures optimal use of the solar spectrum, while the chemically stable metal oxide protects the underlying PV cell from photocorrosion. Recent breakthroughs have brought metal oxide photoelectrodes close to the efficiency levels required for practical applications. We will use our extensive combined expertise on nanostructuring, photon management, and interface engineering to design innovative ways to solve the remaining bottlenecks, and achieve a solar-to-H2 (STH) energy conversion efficiency of 10% for a small area device, with less than 10% performance decrease over 1000 h. In parallel, our academic and industrial partners will collaborate to develop large-area deposition technologies for scale-up to ≥50 cm2. This will be combined with the large-area PV technology already available within the consortium, and used in innovative cell designs that address critical scale-up issues, such as mass transport limitations and resistive losses. The finished design will be used to construct a water splitting module consisting of 4 identical devices that demonstrates the scalability of the technology. This prototype will be tested in the field, and show a STH efficiency of 8% with the same stability as the small area device. In parallel, our partners from industry and research institutions will work together on an extensive techno-economic and life-cycle analysis based on actual performance characteristics. This will give a reliable evaluation of the application potential of photoelectrochemical hydrogen production, and further strengthen Europe's leading position in this growing field.

Bacteria in nature exhibit remarkable capacity to sense their surroundings and rapidly adapt to diverse conditions by gaining new beneficial traits. This extraordinary feature facilitates their survival when facing extreme environments. Utilizing Bacillus subtilis as our primary model organism, we propose to study two facets of this vital bacterial attribute: communication via extracellular nanotubes, and persistence as resilient spores while maintaining the potential to revive. Exploring these fascinating aspects of bacterial physiology is likely to change our view as to how bacteria sense, respond, endure and communicate with their extracellular environment. We have recently discovered a previously uncharacterized mode of bacterial communication, mediated by tubular extensions (nanotubes) that bridge neighboring cells, providing a route for exchange of intracellular molecules. Nanotube-mediated molecular sharing may represent a key form of bacterial communication in nature, allowing for the emergence of new phenotypes and increasing survival in fluctuating environments. Here we propose to develop strategies for observing nanotube formation and molecular exchange in living bacterial cells, and to characterize the molecular composition of nanotubes. We will explore the premise that nanotubes serve as a strategy to expand the cell surface, and will determine whether nanotubes provide a conduit for phage infection and spreading. Furthermore, the formation and functionality of interspecies nanotubes will be explored. An additional mode employed by bacteria to achieve extreme robustness is the ability to reside as long lasting spores. Previously held views considered the spore to be dormant and metabolically inert. However, we have recently shown that at least one week following spore formation, during an adaptive period, the spore senses and responds to environmental cues and undergoes corresponding molecular changes, influencing subsequent emergence from quiescence.

The main objective is the development of highly multiplexed affinity proteomics tools for point of care diagnostics. Enabled by the world's largest resource of antibodies to human protein targets generated within the framework of the Human Protein Atlas program, the aim of this application is to develop new concepts in translational medical research to allow dramatic improvement in performance and accessibility of point of care tests in both high and low resource clinical settings in infectious diseases, autoimmune conditions and several cancers. To achieve the goal, developments will be made in 1) detection principles and sample preparation, 2) rapid and low cost protein microarrays assays and 3) novel amplification strategies. Strategies will be developed to for the first time enable highly multiplexed sandwich detection across the analytical range of the plasma proteome. Four different novel microarray formats amenable for point of care are presented that each will allow dramatically improved multiplexity and high performance. Different types of new multifunctional silica/gold nanoparticles exhibiting combinations of magnetic, fluorescent, electro-active, enzymatic and optical qualities that will allow deterministic actuation and multimodal detection options will be synthesized and applied for exquisite sensitivity and rapid detection. Following the technology development, developed assay systems will be implemented for full plasma proteome analysis and in the three clinical areas: infectious diseases, autoimmune conditions and cancers. Collaborations with clinical researchers in all the relevant fields have already been established. In theses clinical fields, a body of previous research efforts has shown that highly multiplexed plasma analysis may dramatically improve diagnostic accuracy, and it is here that the novel comprehensive point of care systems presented in this proposal may conceivably create the largest impact for patients and health care systems.

Influenza (Flu) virus infections lead to thousands of deaths annually worldwide and billions of dollars of economic burden. Vaccination is the primary strategy for Flu prevention; however, effective protection requires a perfect match with the seasonal circulating strains. Seasonal vaccines target the hemagglutinin (HA) and neuraminidase (NA) proteins of the two predominant A and one B circulating strains, and induce high neutralizing antibody titers. Emerging Flu viruses escape host immunity by modifying HA glycosylation and masking the immunodominant epitopes of previous years' strains. In consequence, yearly seasonal Flu vaccines have to be developed. Therefore, there is a need for a universal Flu vaccine that would protect against most variants. Patterns of HA glycosylation show that there are only 5 -6 alterations in a century of evolution, reflecting the limited possibilities for the virus to escape host immunity while retaining its infectivity. Therefore, by developing a vaccine that includes most relevant past seasonal (H1, H3, and B) and pandemic (H5, H7) strains, it might be feasible to develop a universal Flu vaccine. The project will combine HA selection with the novel self-amplifying mRNA (SAM®) vaccine technology, which consists of a synthetic RNA delivered by lipid nanoparticles, and already proved immunogenic in mice with Flu H1 and H7. We will design five SAM(HA) vectors encoding combinations of four HA, prioritized to cover up to 20 relevant strains, and characterize their immunogenicity in mice by measuring hemagglutination-inhibition and virus neutralization titers. In-depth characterization of germinal center B cells and follicular helper T cells will be performed by mass cytometry (CyTOF). Finally, HA-specific CD4 and CD8 T-cell cytokine and cytotoxic responses will be determined by mass cytometry for correlates of protection. At the end of this proposal, the development of a universal Flu vaccine should be achieved.

The brain is nowadays the object of a number of extensive systematic studies that focus on seemingly all aspects of its morphology and function, from overall brain architecture to neuronal connections, neuronal morphology and gene expression. However, at least one basic aspect is as yet incompletely studied: the molecular anatomy of the neuron, i.e., the copy numbers and the spatial arrangement of molecules within the neuronal cell. This cannot be addressed by gene expression or proteomics approaches, as they do not have sufficient spatial precision. Electron microscopy, ideal for unraveling the neuronal morphology, does not have sufficient protein labeling efficiency. My project aims to fill this gap by a combination of super-resolution fluorescence microscopy, advanced fluorescence labelling techniques and advanced biochemistry tools such as quantitative mass spectometry. My objectives are fourfold: 1) to determine the molecular organization of at least 200 major neuronal proteins: their exact copy numbers and their position within the cell determined with nanoscale precision; 2) to generate a bank of biochemistry and microscopy sample preparations that will be available world-wide for the characterization of 1000-2000 additional proteins; 3) to integrate results into an in silico neuronal model that can be used for modelling functional neuronal parameters; 4) to use this technology to determine the changes in neuronal anatomy caused by neurodegeneration in Alzheimer's and Parkinson's Diseases. My group has already performed key preliminary work towards these aims. Our preliminary work focused on the synapse, where we ascertained copy numbers and positions for proteins adding up to more than 50% of the synapse's protein mass (see figure). We are thus ready to embark on this large-scale, risk-taking project. We are confident that the in silico neuron and the preparation bank we will create will represent key new resources for future studies in neurobiology.

This proposal aims the design of novel optical enhancers for surface-enhanced Raman scattering (SERS) to develop routine methods for quantitative detection of a very broad range of substances based on regular arrays of gold nanorods. As proof of concept the diagnosis of neurodegenerative diseases, such as Alzheimer or Creutzfeldt-Jakob diseases (CJD), which still pose a great challenge for international health systems because of the economic and social impact of its pandemic outbreaks. Such diseases do not induce any immunological response on the infected individuals and thus, antibody detection cannot be used because they are not produced by the host. Therefore, antibody-free detection systems are required, which are also highly sensitive and selective. Overall this presents a significant challenge which can be resolved using a new generation of SERS substrates with unprecedented degree of structural control. Therefore, the main objectives of the project will involve the fabrication of uniform gold nanorods within a wide range of sizes; their assembly into perfectly ordered supercrystals, both on planar and patterned substrates; evaluation of the SERS enhancing properties of such assemblies as a function of nanorod morphology and degree of order; comparison with theoretical modeling and prediction of the most convenient configuration; and finally the full implementation of the detection of prions using these substrates. We propose the use of SERS as a non-invasive sensor, which is able to detect and monitor prions in biological fluids (blood, urine or saliva). For this target to be achieved, the enhancing metallic substrates need to be engineered for focalization of the plasmonic modes at certain regions of the substrate and production of extremely high enhancement factors. This idea is based on the concept of field localization by nanoantennas, which will be perfectly applicable to the oriented gold nanorod colloidal crystals.

Development of new strategies to treat neurodegenerative diseases is one of the key priorities of the European Union. Their socioeconomic burden is rapidly growing due to the increasing lifespan and the decreasing percentage of working population, currently costing the EU €130 billion a year in care. Despite efforts put in development of treatments for neurodegeneration, the bench-to-bedside translation of neuroprotective strategies remains very low. Major factors contributing to this problem are incomplete understanding of the mechanisms behind neuronal injuries, lack of compounds affecting multiple protective pathways/cell types, and side effects caused by broad-spectrum neuroprotectants. A separate crucial problem is the limited blood–brain barrier (BBB) passage of most compounds. Therefore, identification of effective therapeutic targets in the brain and delivery of novel low-cost neuroprotectants with minimal side effects and high BBB passage is of paramount importance. The emergence of nanoneuroscience is revolutionizing treatment of CNS disorders. This approach uses nanometer-scale materials, which can interact with biological systems at a molecular level, bypass cellular barriers and induce desired physiological responses in cells with minimal side effects. The project proposed here combines advanced methods of nanoscience and neurobiology to (1) characterize novel therapeutic targets and neuroprotective pathways in the brain and (2) design novel efficient nanoprotectants against neurodegenerative conditions, such as stroke, epilepsy, Parkinson's and Alzheimer's diseases. The project will offer exemplary training for the Fellow in nanoneuroscience, as well as in project management skills, at Imperial College, ranked the 5th highest university internationally. These competencies will complement the already impressive range of the Fellow's research and managing capabilities and will help her to develop as a leader in this frontier area of science.

The success of modern medical treatments such as cellular therapy and targeted treatments requires appropriate tools for in vivo monitoring. Imaging modalities, such as magnetic resonance imaging (MRI), single photon emission computed tomography (SPECT) and positron emission tomography (PET) are key candidates due to their noninvasive nature. However, these imaging techniques are extremely expensive and can involve radiation, both of which hinder their longitudinal and repetitive use. Ultrasound has so far been unsuitable due to the absence of a label to differentiate regions of interest from tissue background, the main problem being that current ultrasound contrast agents (CAs) have active lifetimes in the order of minutes. The CoNQUeST platform (Clinical Nanoparticles for Quantitative Ultrasound with high STability) proposed here is an entirely new type of ultrasound CA that is extremely stable (lifetime of a year) and is not affected by insonation. This mechanism of contrast generation appears completely novel: The polymeric particles are under 200nm in diameter and must contain a soluble metal (M.Srinivas et al., patent pending, filed 09/2012). Based on the current state of the art, these particles are too small and do not contain the requisite gaseous component for ultrasound contrast. CoNQUeST particles are applicable to longitudinal and repeated imaging, as is necessary for cell tracking, due to their stability. Furthermore, these particles can be chemically bound to targeting agents, dyes and drugs, and are suitable for multimodal imaging, including MRI (both 1H and 19F), fluorescence and SPECT. Finally, the CoNQUeST agents are suitable for clinical use. I propose the application of the CoNQUeST agents to a clinical trial for tracking dendritic cell therapy in melanoma patients, longitudinal theranostic imaging in preclinical models and thorough characterisation of this novel mechanism of ultrasound contrast generation.