Laser and optical based methods for detecting and characterising microorganisms

Infectious diseases continue to kill millions of people every year and are a significant burden on the socio-economic development of developing countries. After many years of international policy aimed at containing diseases, it has recently become an explicit aim to move towards elimination of infectious diseases. However, if this is to occur, it will be necessary to have highly eficacious diagnostic tools to ensure infected individuals are identified and treated. However, the diagnosis of infectious diseases in the developed and developing world requires the full integration of complex assays in easy-to-use platforms with robust analytical performances at low cost. Many relevant bioanalytical technologies have been developed for use in laboratories and clinics, including the current gold standard for the diagnosis of tuberculosis and malaria. The miniaturization and integration of complex functions into lab-on-a-chip (LOC)technologies using microfluidics have only had limited success in translating diagnosis assays out of a centralized laboratory to point-of-care (POC) settings, because they still remain constrained due to chip interconnection and they are either not likely to go out of research laboratories or are not appropriate for low resource settings. In this thesis, a new microfluidic platform was developed that reduced the dependency of the diagnostic procedure on large laboratory instruments providing simplicity of use, enabling the patient sample to be processed and diagnosed on a low cost, disposable biochip. Surface acoustic wave (SAW) devices, which are commonly used in mobile phone technologies, were adapted to provide controlled microfluidic functions by shaping the SAW using particular designs of electrodes and phononic structures. The control of lateral positioning of the SAW was demonstrated using a slanted finger interdigitated transducer (IDT) in a frequency tuneable manner allowing microfluidic functions such as mixing, moving and merging, sequentially performed using a single IDT both on the substrate and on a disposable chip. Alternatively, phononic bandgaps were designed to break the symmetry of the SAW in a tuneable manner and gradient index phononic crystals (GRIN-PC) lenses were designed to focus the SAW and successfully increased the amplitude of the wave by a factor 3 while the focal position could be tuned with the frequency. The potential of these techniques was demonstrated by controlling the amplitude and direction of water jet towers by the use of a phononic horn structure that allowed the enhancement of energy at defined positions and by propelling and directing a macrometer scale object in water using a slanted IDT. As proof of concepts of diagnostic devices for the developing world, an immunoassay for tuberculosis using only mobile phone technologies (SAW, light-emitting diode(LED) and complementary metaloxidesemiconductor (CMOS) camera) was demonstrated with a limit of detection of 1 pM, which is the limit required in an interferon-release assay. This limit of detection was only achievable because of the ability of SAW to increase the mixing and to reduce the non-specific binding. Furthermore, a method to enrich malaria infected cells, based on SAW and isopycnic gradient, was also demonstrated and showed an enrichment up to 100x in the equivalent of a fingerprick of blood in 3 seconds. This technique will allow to reduce the limit of detection of the current gold standard. This platform not only opens a clear road toward POC diagnostics due to its size, cost, versatility and ease in integration, but has also the potential to provide useful tools in laboratory settings for large scale, high throughput technologies.