Ischaemic heart disease is a major healthcare issue, resulting globally in 12.2% of all deaths, but rising to a massive 16.3% amongst the wealthiest countries. This makes it the leading cause of death and with an aging population this is expected to remain a major issue. Bio-medical research is at the forefront of the movement to understand the onset of heart conditions, trauma and their treatment, and includes detailed studies at the cellular level. Pivotal in these studies is the ability to simulate these conditions and examine the membrane integrity and viability of the cardiac myocyte, the contractile cell from the heart. This project will develop a tool to facilitate these studies using acoustically induced fluid flows which produce sub-millimeter sized vortex flows, termed ultrasonic micro-streaming , to generate shear forces within the cell membrane; by introducing and manipulating micro-streaming sources such as micro-bubbles, it will be shown how ultrasonic streaming can be targeted at isolated cardiac myocyte cells to either test their integrity or induce a controlled level of trauma to test subsequent cardioprotective treatments and restoration of cell function.

We have carefully planned this research programme to pioneer a wholly new capability in ultrasonic particle manipulation to allow electronic sonotweezers to take their place alongside optical tweezers, dielectrophoresis and other techniques in the present and future particle manipulation toolkit.Following end-user demand, particle manipulation is a rapidly growing field, notably applied to the life sciences, with emerging applications in analysis and sorting, measurement of cell forces and tissue engineering. Existing devices have valuable capabilities but also limits in terms of forces that can be produced and measured, particle sizes that can be handled, their range of compatible buffer characteristics and sensitivity to heating, and suitability for integration with sensors in low cost devices. Key to our programme is the concept of dynamic potential energy landscapes and the established ability of ultrasound to create such landscapes, potentially to generate forces under electronic / computer control. Our principal technical aim is to exploit this in integrated sonotweezers to apply and measure larger forces over longer length scales, extend micromanipulation to larger particles, and demonstrate this in pathfinder applications in life sciences.To achieve our aims, we have already carried out successful feasibility studies and brought together an outstanding multidisciplinary team of investigators including internationally established members, some of the UK's most exciting young scientists and engineers, and appropriate overseas collaborators. Such a team is a prerequisite for what we recognise as a challenging, highly complex, densely interlinked programme. Over its four years, with strong management and built-in research flexibility, we will explore key areas of science, technology and applications to create and demonstrate electronic sonotweezers. Throughout the work, there will be parallel activity in understanding of physical principles, modelling and design, state-of-the-art fabrication, sensor integration, and applications testing.

A major challenge in biology is to understand how cells recognize external signals and give appropriate responses. Now that the sequence of the human genome is complete, it is important to assign functions to each gene and to identify the corresponding proteins that control key cellular functions. White and colleagues pioneered the development of microscopy-based methods for the visualization and timelapse measurement of biological processes in single living cells. We have used natural light-emitting proteins from fireflies, jelly fish and fluorescent corals. Synthesis (expression) of these proteins causes mammalian cells to become luminescent (light emitting in the dark) or fluorescent (change the colour of light). By placing the gene that codes for a luminescent protein next to a promoter that controls a gene of interest, we can use luminescence from living cells as a way of measuring when the gene of interest is normally switched on and off. Fluorescent proteins have also been used to genetically label proteins of interest, so that the movement of the protein can be visualized in a living cell. White and colleagues previously used timelapse fluorescence and luminescence microscopy coupled to computer simulations to investigate cell decision making. We discovered that a set of important signalling proteins, called NF-kappaB, move repeatedly into and out of the nucleus of the cell, suggesting that cells may use proteins as timers to encode complex messages (like Morse Code). This was a surprise since the original NF-kappaB protein, p65, was discovered 20 years ago and was thought to act as a simple switch that moves into the nucleus once to activate genes. Only timelapse measurements in single living cells were able to see this. The NF-kappaB system is widely recognised as crucial to the control of important cellular processes including both cell division and cell death. It is implicated as being involved in a variety of diseases, such as cancer and inflammatory disease. We will now develop a substantial systems biology project to study all of the components of this complex system. While the previous work has provided major insights, we now need a far broader range of integrated experimental tools to study it. Also the use of mathematical models to make computer predictions will be critical to help us to visualize how this system works. We will make accurate measurements of the (much larger) set of proteins that are involved in NF-kappaB signalling and the genes that are controlled by these signals. The (very experienced) project team includes bioinformaticians, cell biologists, computer scientists, mathematicians, molecular biologists, microscopists and protein chemists. The project will be managed in a structured and organized way, so that the mathematical modelling can be used to predict and design the biological experiments. A central team of experimental officers will be responsible for coordinating the experiments, data and model storage and communication of information between team members. We will study the numbers of molecules of each of the NF-kappaB proteins in the cell, their stability, chemical states and interactions with each other and with other proteins. We will also study in detail which genes that they bind to and control. We will also aim to understand how single protein molecules acting at single genes can act to control decisions of cell life and death. This multidisciplinary approach is essential in order to understand this complex system. A further aim of the project is to provide training for post-docs and students. In this respect, we will benefit from sponsorship of training courses and symposia by the instrumentation companies Carl Zeiss, Hamamatsu Photonics, Coherent and Nano Imaging Devices. The project will also benefit from ongoing collaborations with Genetix and AstraZeneca

We have carefully planned this research programme to pioneer a wholly new capability in ultrasonic particle manipulation to allow electronic sonotweezers to take their place alongside optical tweezers, dielectrophoresis and other techniques in the present and future particle manipulation toolkit.Following end-user demand, particle manipulation is a rapidly growing field, notably applied to the life sciences, with emerging applications in analysis and sorting, measurement of cell forces and tissue engineering. Existing devices have valuable capabilities but also limits in terms of forces that can be produced and measured, particle sizes that can be handled, their range of compatible buffer characteristics and sensitivity to heating, and suitability for integration with sensors in low cost devices. Key to our programme is the concept of dynamic potential energy landscapes and the established ability of ultrasound to create such landscapes, potentially to generate forces under electronic / computer control. Our principal technical aim is to exploit this in integrated sonotweezers to apply and measure larger forces over longer length scales, extend micromanipulation to larger particles, and demonstrate this in pathfinder applications in life sciences.To achieve our aims, we have already carried out successful feasibility studies and brought together an outstanding multidisciplinary team of investigators including internationally established members, some of the UK's most exciting young scientists and engineers, and appropriate overseas collaborators. Such a team is a prerequisite for what we recognise as a challenging, highly complex, densely interlinked programme. Over its four years, with strong management and built-in research flexibility, we will explore key areas of science, technology and applications to create and demonstrate electronic sonotweezers. Throughout the work, there will be parallel activity in understanding of physical principles, modelling and design, state-of-the-art fabrication, sensor integration, and applications testing.

We have carefully planned this research programme to pioneer a wholly new capability in ultrasonic particle manipulation to allow electronic sonotweezers to take their place alongside optical tweezers, dielectrophoresis and other techniques in the present and future particle manipulation toolkit.Following end-user demand, particle manipulation is a rapidly growing field, notably applied to the life sciences, with emerging applications in analysis and sorting, measurement of cell forces and tissue engineering. Existing devices have valuable capabilities but also limits in terms of forces that can be produced and measured, particle sizes that can be handled, their range of compatible buffer characteristics and sensitivity to heating, and suitability for integration with sensors in low cost devices. Key to our programme is the concept of dynamic potential energy landscapes and the established ability of ultrasound to create such landscapes, potentially to generate forces under electronic / computer control. Our principal technical aim is to exploit this in integrated sonotweezers to apply and measure larger forces over longer length scales, extend micromanipulation to larger particles, and demonstrate this in pathfinder applications in life sciences.To achieve our aims, we have already carried out successful feasibility studies and brought together an outstanding multidisciplinary team of investigators including internationally established members, some of the UK's most exciting young scientists and engineers, and appropriate overseas collaborators. Such a team is a prerequisite for what we recognise as a challenging, highly complex, densely interlinked programme. Over its four years, with strong management and built-in research flexibility, we will explore key areas of science, technology and applications to create and demonstrate electronic sonotweezers. Throughout the work, there will be parallel activity in understanding of physical principles, modelling and design, state-of-the-art fabrication, sensor integration, and applications testing.