Ines Frese

Microfluorimeter with disposable polymer chip for detection of coeliac disease toxic gliadin

Coeliac disease is an inflammatory disease of the upper small intestine and results from gluten ingestion in genetically susceptible individuals, and is the only life-long nutrient-induced enteropathy. The only treatment is a strict gluten-free diet and the longer the individual fails to adhere to this diet, the greater the chance of developing malnutrition and other complications. The existence of reliable gluten free food is crucial to the well-being of the population. Here we report on a microfluorimeter device for the in situ detection of gliadin in foodstuffs, which could be used for a rapid control of raw materials in food processing, as well as for process control of gliadin contamination. The microfluorimeter is based on a reflector that is used inside a microfluidic chip, exploiting various strategically placed reflective or totally metallised mirrors for efficient collection of the fluorescent light emitted in a large solid angle. The chip is capable of executing five assays in parallel and has been demonstrated to possess detection sensitivity applicable to fluoroimmunoassays. Various immunoassay formats exploiting fluorescence detection, using enzyme/fluorophore labels were developed and compared in terms of sensitivity, ease of assay, assay time and compatibility with buffer used to extract gliadin from raw and cooked foodstuffs, with the best performance observed with an indirect competition assay using a fluorophore-labelled anti-mouse antibody. This assay was exploited within the microfluorimeter device, and a very low detection limit of 4.1 ng/mL was obtained. The system was observed to be highly reproducible, with an RSD of 5.9%, for a concentration of 50 ng/mL of gliadin applied to each of the five channels of the microfluorimeter. Biofunctionalised disposable strips incorporated into the microfluorimeter were subjected to accelerated Arrhenius thermal stability studies and it was demonstrated that strips pre-coated with gliadin could be stored for approximately 2 years at 4 degrees C, with no discernable loss in sensitivity or detectability of the assay. Finally, the microfluorimeter was applied to the analysis of commercial gluten-free food samples, and an excellent correlation with routine ELISA measurements was obtained. The developed microfluorimeter should find widespread application for on-site execution of fluoroimmunoassays.

Plastic micro-optical components with the LIGA technology

Moulding of plastics enables optical features to be integrated into a single unit. This is particularly an advantage for product designs that impose space and weight constraints. Therefore, the use of plastic for biomedical and non telecommunications orientated optical applications continues to grow as design engineers take advantage of the ease of fabrication and the material flexibility. Deep X-ray LIGA presents itself as a method ideally suited for the production of moulds for the manufacture of plastic microcomponents. LIGA is synonymous for the lithography preferably carried out with synchrotron radiation X-rays, although many other lithography and non-lithography methods for master production have been developed in the last few years. Nevertheless, the exceptional resist heights, the enormous accuracy and low runout as well as the low sidewall roughnesses cannot be copied by these other methods of master production. In particular, the low sidewall roughnesses achieved through deep X-ray LIGA is essential for the manufacture of waveguide coupling systems based on polymers. The design and conceptualisation of such waveguides systems is presented here. In addition however, the exceptional resist heights and low runout can be employed to produce passive structures for the packaging of optical components. This paper provides an overview of the deep X-ray LIGA technology, emphasizing its strengths and application areas. Considerations for the design and manufacture of the plastic structures are also elucidated.

Water analysis in a lab-on-a-chip system

The development of a lab-on-chip system which allows the parallel detection of a variety of different parameters of a water sample is presented. Water analysis typically comprises the determination of around 30 physical and chemical parameters. An even larger number can arise when special contaminations of organic molecules are of interest. A demonstration system has been realised to show the feasibility and performance of an integrated device for the determination of physical quantities like electrical conductivity, light absorption and turbidity. Additionally, chemical quantities like the pH-value and the content of inorganic and organic contaminations are also determined. Two chips of credit card size contain the analytical functions and will be fabricated by injection moulding. First prototypes have been manufactured by milling or precision milling for the optical components.