DNA profiling represents one of the most important and growing techniques currently use in the UK to investigate and fight crime. Established in 1995 the UK data base, which is the world's largest per head of population, currently holds just under 5 million individual profiles, based on short repeating DNA sequences, offering a discriminating power of one in 50 million. At present DNA profiling is being used to solve around 50,000, or one in every 300 recorded crimes per year, in the UK. The effectiveness and use of the technique in supporting police investigations could however be significantly improved if the unit cost and time taken from collecting a sample to obtaining a profile could be shortened from the current days to hours. In July 2006 an ambitious three year project, funded by the EPSRC under its 'Think Crime' initiative (EP/D040930), was undertaken at the University of Hull to develop a micro fluidic based Lab-on-a-chip system which could be used to obtain DNA profiles at the scene of a crime. The work has seen the successful design and fabrication of micro fluidic chips and associated control system, that is able to extract DNA from a range of sample types (note cell lysis performed off chip to release DNA), perform PCR amplification of a multiplex sample and then separate and detect products in approximately 2 hours. The chip used to perform this analysis has no moving parts (reagents and sample are pumped by electric fields) and contains all the reagents in gels on chip, avoiding the possibility of contamination as once the sample is placed on the chip and sealed nothing enters or leaves the device. The micro fluidic device which is the subject of a UK patent application submitted in October 2008 (Application No. 0818609.0) is made of glass (120mm long, 60mm wide and 2 mm deep) and fits into a custom built instrument, about the size of a domestic microwave (60cm long, 30cm wide and 40 cm high), that controls the generation of the DNA profile (movement, amplification and separation/detection). However having reached the proof of principle stage in the current project the applicants are now very keen to make the technology accessible to the police in their fight against crime.Having carried out an initial market and IP evaluation in conjunction with the Forensic Scientific Service the idea of adapting the methodology to provide a custody suite based technology, allowing DNA profiling in under two hours from a buccal (mouth) swab sample, has been identified as a significant need which is realistically achievable following minor modifications to the current control instrument and chips. The DNA profile would be achieved at a competitive price due to the very small amounts of reagents required. An initial evaluation of market competitors and potentially blocking IP has indicated that a good opportunity exists to develop the proposed technology which will address a unique market slot. Accordingly funding will be used to support the technical modifications required to convert the current technology into a demonstrator system for custody based use. The Follow-on proposal will also allow retention of key research personnel with specialised practical skills and technical knowhow, who will gain additional skills in commercialisation and product development. Furthermore a more comprehensive freedom to operate study and business plan will be produced to carry the work forward into a full commercially-based activity. In the context of this proposal it is important to stress that given the current strong emphasis being place on a growing DNA database the UK represents an ideal location in which to launch the proposed new technology. It is clear also that internationally, DNA fingerprinting and biometric analysis represents an emerging field and early market dominance will clearly offer the potential to develop a major UK product base to address a growing global arena for forensic and related applications.

Forensic investigations frequently involve the analysis of body fluids spilled during the commissioning of a crime, leading to deposits on a wide range of surfaces such as carpets, walls, clothing, bedlinen and skin. Testing of, for example, bloodstains for the presence of drugs and drug-related metabolites or other biomarkers can be a highly complex procedure, and the success rate of such analyses can be severely limited due to loss of material during extraction, as well as a lack of absolute sensitivity. Moreover, the inherently destructive nature of these tests - specimens need to be removed from the bulk item before any extraction can be carried out - is a major issue, as is the fact that analysis is invariably carried out at a laboratory remote from the scene. The necessity for remote analysis does also add potentially crucial time to the investigative process, as well as carrying an inherent risk of sample degradation and contamination in transit and subsequent storage. Perhaps most importantly: if it is not feasible to remove a suitable sample from an item, potentially useful information will not be available to investigating officers. Therefore, forensic practitioners have considerable interest in technologies which could potentially overcome at least some of these important limitations.In this proposal we intend to undertake a feasibility study to assess a novel application of the established technique of mass spectrometry in such forensic investigations. Mass spectrometry is a very powerful and well-established technique able to detect minute amounts of materials with very high sensitivity and specificity. Unfortunately, it has some serious drawbacks which severely restrict its applicability. These drawbacks chiefly concern the sampling of the materials under investigation, which have to be capable of being in the gas phase. This tends to be especially difficult from solid substrates, necessitating physical removal and dissolution of the material of interest, bombardment of the sample with highly energetic ions in an ultrahigh vacuum chamber, or illumination of the sample with an intense laser beam after application of a matrix material. All these methods are highly problematic in the field of forensic analysis, where integrity of the subject is of utmost importance. A methodology to volatilise materials of interest for mass spectrometric analysis which is compatible with forensic requirements would thus have great potential.The newly-introduced technique of DESI, short for desorption electrospray ionisation, has such potential, as yet to be realised, to become a fully-fledged routine surface analytical tool, marrying the analytical capabilities of mass spectrometry with a sampling facility convenient for forensic analyses. As opposed to most other such techniques, it is applicable under a wide range of environmental conditions and suitable for a wide array of samples including surfaces of biological, pharmaceutical, polymer, metal and mineral materials. Moreover, current technical capabilities are such that a great deal of miniaturisation is possible, leading to a desktop-sized or potentially even a portable device.The proposed work focuses on the application of the technique to forensically relevant subjects and materials, more specifically the detection of drugs and drug metabolites in body fluid stains found at scenes of crime. Direct comparisons will be made with mass spectra generated using two established mass spectrometry techniques capable of surface analysis available in the School of Pharmacy, secondary ion mass spectrometry (SIMS) and matrix assisted laser desorption-ionisation (MALDI), which both employ different methods of volatilisation. It should be stressed that whereas this proposal concentrates on a very specific sample type, the technique has the clear potential for application to a much wider range of samples in the forensic science field.

Forensic science is an important tool in the fight against crime and this proposal will build on cutting edge research to create technology that will revolutionise the way DNA fingerprinting can be used at crime scenes to generate quick and accurate answers where they are urgently needed. Over the past decade DNA fingerprinting, together with the establishment of a national database, has becoming one of the most important forensic tools in the fight against crime, with its unique analytical capability to biometrically profile suspects. At present such analysis is carried out in the laboratory environment which requires samples to be collected from the scene of a crime, and then be transported to a central facility for subsequent analysis. This current practice clearly adds additional transport and storage time which in turn increase costs and adds unwanted delays in analysis times. Therefore the development of field-based methodology could prove to be most valuable in generating more rapid DNA-based intelligence. Such capability will not only allow DNA profiles to be obtained in say less than 30 minutes but the parallel operation of such methodology will allow reasonably large numbers of samples to be run before the crime scene becomes contaminated or corrupted. Whilst the basic analytical methodology required to carry out DNA analysis is well established (i.e. extraction and purification of the DNA from the biological sample, amplification of the target DNA by polymerase chain reaction (PCR) and electrophoretic size separation and fluorescent detection of the DNA fragments) the development of a portable device which integrates these hitherto separate functions is a unique challenge which form the basis of this proposal.In order to achieve true hand-held portability the technology needs to be mechanically and chemically/biochemically robust and have low power requirements for battery-based operation. In this proposed project the combined expertise of the applicants will be focused on generating a prototype instrument that will meet the portability requirements outlined above. The work builds on early encouraging research in which micro fluidic methodology coupled with efficient microwave-based heating has been demonstrated to be suitable for the PCR amplification of DNA samples. The micro fluidic approach, which offers small sample capability (microlitres), has already been shown by number of researchers, including the applicants, to offer a rapid approach to the thermal PCR cyclic process suggesting realistic processing times of around 20 minutes. Following amplification of the target DNA, analysis can effectively be performed using capillary electrophoresis-based separations with fluorescence detection and once again the micro fluidic approach has proved advantageous in offering an efficient and rapid separation process within a few minutes. The prototype developed in this application will be based on an integrated micro fluidics manifold or chip that will enable sample extraction/preconcentration, PCR amplification and DNA fragment separation to be achieved whilst exploiting low power requirements by using manual and electrokinetic fluidic pumping, low power solid state microwave-based heating and low power fluorescent detection. Whilst all of the sample processing stages indicated have been demonstrated separately, it is important to stress that there is a real scientific challenge in bringing them all together in an integrated unit with compatible functions and requirements at each stage. If successful, the product will be an example of novel low-cost but highly functional technology, which will be of immediate benefit in the fight against crime. We know this is not going to be easy but we believe the research team assembled around this proposal can get the job done and break the integration barrier to success that currently exists.

People involved in trying to find missing persons have to depend upon images of them taken before they disappeared. For missing adults, this is not too great a problem, as their appearance will not have changed radically. However, children may have changed out of all recognition. The aim of this project is to develop a way of 'ageing' a facial image to give a realistic impression of what the missing child will look like now. If successful, the system will be a major boost to the work of those locating missing children: it will provide accurate images rapidly and simply at a fraction of the current cost.The project will develop algorithms, or step-by-step problem-solving procedures, that can be used to age a facial image. These will be based on studies of facial variation in children over two critical stages in their development which will be undertaken at Dundee University, and from existing face data archives held at the University of Kent. The project will benefit from having very close interaction and guidance from the National Missing Persons Helpline (NMPH) and VisionMetric, a company that has a good deal of commercial experience in developing forensic imaging applications.The final outcome of the project will be a software system that can be used on standard hardware and over the Internet. To get to this stage, however, there will be a series of project stages:Compiling and annotating a photographic database of children's faces spanning two main developmental phases (at Dundee)Developing a comprehensive statistical model of facial appearance (at Kent)Developing person-specific age transformation techniques (at Dundee and Kent)Developing these techniques to make them quick and user-friendly, so that they can be used for practical or commercial development (at Kent)Practical implementation of the software system (at Kent, involving VisionMetric, NMPH and the Metropolitan Police)

People involved in trying to find missing persons have to depend upon images of them taken before they disappeared. For missing adults, this is not too great a problem, as their appearance will not have changed radically. However, children may have changed out of all recognition. The aim of this project is to develop a way of 'ageing' a facial image to give a realistic impression of what the missing child will look like now. If successful, the system will be a major boost to the work of those locating missing children: it will provide accurate images rapidly and simply at a fraction of the current cost.The project will develop algorithms, or step-by-step problem-solving procedures, that can be used to age a facial image. These will be based on studies of facial variation in children over two critical stages in their development which will be undertaken at Dundee University, and from existing face data archives held at the University of Kent. The project will benefit from having very close interaction and guidance from the National Missing Persons Helpline (NMPH) and VisionMetric, a company that has a good deal of commercial experience in developing forensic imaging applications.The final outcome of the project will be a software system that can be used on standard hardware and over the Internet. To get to this stage, however, there will be a series of project stages:Compiling and annotating a photographic database of children's faces spanning two main developmental phases (at Dundee)Developing a comprehensive statistical model of facial appearance (at Kent)Developing person-specific age transformation techniques (at Dundee and Kent)Developing these techniques to make them quick and user-friendly, so that they can be used for practical or commercial development (at Kent)Practical implementation of the software system (at Kent, involving VisionMetric, NMPH and the Metropolitan Police)