Automated Particle Identification through Regression Analysis of Size, Shape and Colour

Rapid point of care diagnostic tests and tests to provide therapeutic information are now available for a range of specific conditions from the measurement of blood glucose levels for diabetes to card agglutination tests for parasitic infections. Due to a lack of specificity these test are often then backed up by more conventional lab based diagnostic methods for example a card agglutination test may be carried out for a suspected parasitic infection in the field and if positive a blood sample can then be sent to a lab for confirmation. The eventual diagnosis is often achieved by microscopic examination of the sample. In this paper we propose a computerized vision system for aiding in the diagnostic process; this system used a novel particle recognition algorithm to improve specificity and speed during the diagnostic process. We will show the detection and classification of different types of cells in a diluted blood sample using regression analysis of their size, shape and colour. The first step is to define the objects to be tracked by a Gaussian Mixture Model for background subtraction and binary opening and closing for noise suppression. After subtracting the objects of interest from the background the next challenge is to predict if a given object belongs to a certain category or not. This is a classification problem, and the output of the algorithm is a Boolean value (true/false). As such the computer program should be able to ”predict” with reasonable level of confidence if a given particle belongs to the kind we are looking for or not. We show the use of a binary logistic regression analysis with three continuous predictors: size, shape and color histogram. The results suggest this variables could be very useful in a logistic regression equation as they proved to have a relatively high predictive value on their own

Creating tissue on chip constructs in microtitre plates for drug discovery

We report upon a novel coplanar dielectrophoresis (DEP) based cell patterning system for generating transferrable hepatic cell constructs, resembling a liver-lobule, in culture. The use of paper reinforced gel substrates provided sufficient strength to enable these constructs to be transfered into 96-well plates for long term functional studies, including in the future, drug development studies. Experimental results showed that hepatic cells formed DEP field-induced structures corresponding to an array of lobule-mimetic patterns. Hepatic viability was observed over a period of 3 days by the use of a fluorescent cell staining technique, whilst the liver specific functionality of albumin secretion showed a significant enhancement due to the layer patterning of cell lines (HepG2/C3A), compared to 2D patterned cells and un-patterned control. This “build and transfer” concept could, in future, also be adapted for the layer-by-layer construction of organs-on-chip in microtitre formats

Continuous cell lysis in microfluidics through acoustic and optoelectronic tweezers

A versatile platform for efficient cell lysis using a combination of acoustic and electric fields in a microchannel is presented. Cell membrane disruption is triggered by electric fields inducing electroporation and then lysis. The principle of optoelectronic tweezers (OET) is applied to control the electric field strength and a surface acoustic wave transducer is attached to an OET chip to implement acoustic tweezing (AT). The system is characterized in terms of spatial control of electric fields, single cell precision and lysi

Acoustically controlled enhancement of molecular sensing to assess oxidative stress in cells

We demonstrate a microfluidic platform for the controlled aggregation of colloidal silver nanoparticles using surface acoustic waves (SAWs), enabling surface enhanced Raman scattering (SERS) analysis of a cell based model for oxidative damage. We show that by varying the frequency and the power of the acoustic energy, it is possible to modulate the aggregation of the colloid within the sample and hence to optimise the SERS analysis

Biocompatibility of a lab-on-a-pill sensor in artificial gastrointestinal environments

n this paper, we present a radiotelemetry sensor, designed as a lab-in-a-pill, which incorporates a two-channel microfabricated sensor platform for real-time measurements of temperature and pH. These two parameters have potential application for use in remote biological sensing (for example they may be used as markers that reflect the physiological environment or as indicators for disease, within the gastrointestinal tract). We have investigated the effects of biofouling on these sensors, by exploring their response time and sensitivity in a model in vitro gastrointestinal system. The artificial gastric and intestinal solutions used represent a model both for fasting, as well as for the ingestion of food and subsequent digestion to gastrointestinal chyme. The results showed a decrease in pH sensitivity after exposure of the sensors for 3 h. The response time also increased from an initial measurement time of 10 s in pure GI juice, to ca. 25 s following the ingestion of food and 80 s in simulated chyme. These in vitro results indicate that changes in viscosity in our model gastrointestinal system had a pronounced effect on the unmodified sensor

Effects of organic and ‘low input’ production methods on food quality and safety

The intensification of agricultural production in the last century has resulted in a significant loss of biodiversity, environmental problems and associated societal costs. The use of shorter rotations or monocropping and high levels of mineral fertilisers, pesticides and crop growth regulators may also have had negative impacts on food quality and safety. To reverse the negative environmental and biodiversity impacts of agricultural intensification, a range of different ‘low input’ farming systems have been developed and are now supported by EU and government support schemes. A range of recent reviews concluded that switching to low input, integrated or organic farming practices results in significant environmental benefits and increased biodiversity in agro-ecosystems. Some recent studies also reported higher levels of nutritionally desirable compounds (e.g. vitamins, antioxidants, mineral nutrients) in foods from organic and ‘low input’ production systems compared to food from conventional systems. The increasing demand and current price premiums achieved by foods from low input and especially organic production systems were shown to be closely linked to consumer perceptions about nutritional and health benefits of such foods. However, there are other studies reporting no significant differences in composition between low input and conventional foods, or inconsistent results. There is currently a lack of (a) factorial studies, which allow the effect of individual production system components (e.g. rotation design, fertility management, crop health management, variety choice) on food composition to be assessed and (b) dietary intervention or cohort studies which compare the effect of consuming foods from different production systems on animal and/or human health. It is therefore currently not possible to draw overall conclusions about the effect of low input production on food quality and safety. This paper will (a) describe the range of organic and other ‘low input’ standards, certification and support systems currently used, (b) summarise the currently available information on effects of organic and other low input crop production systems on the environment, biodiversity and food quality, and (c) describe the methodologies and results from subproject 2 of the EU-funded Integrated project QualityLowInputFood. This project focused on improving our knowledge about the effect of organic and low input crop and livestock production systems on food quality and safety parameters

A programmable microsystem using system-on-chip for real-time biotelemetry

A telemetry microsystem, including multiple sensors, integrated instrumentation and a wireless interface has been implemented. We have employed a methodology akin to that for System-on-Chip microelectronics to design an integrated circuit instrument containing several "intellectual property" blocks that will enable convenient reuse of modules in future projects. The present system was optimized for low-power and included mixed-signal sensor circuits, a programmable digital system, a feedback clock control loop and RF circuits integrated on a 5 mm × 5 mm silicon chip using a 0.6 μm, 3.3 V CMOS process. Undesirable signal coupling between circuit components has been investigated and current injection into sensitive instrumentation nodes was minimized by careful floor-planning. The chip, the sensors, a magnetic induction-based transmitter and two silver oxide cells were packaged into a 36 mm × 12 mm capsule format. A base station was built in order to retrieve the data from the microsystem in real-time. The base station was designed to be adaptive and timing tolerant since the microsystem design was simplified to reduce power consumption and size. The telemetry system was found to have a packet error rate of 10<sup>-</sup><sup>3</sup> using an asynchronous simplex link. Trials in animal carcasses were carried out to show that the transmitter was as effective as a conventional RF device whilst consuming less power

Effect of organic, low-input and conventional production systems on pesticide and growth regulator residues in wheat, potato and cabbage

The Nafferton factorial systems comparison (NFSC) experiments facilitate the investigation of effects of, and interaction between, three production system components - a) rotational position, b) fertility and c) crop protection management - in organic, conventional and low-input crop management systems. This paper presents first results on pesticide and growth regulator residues observed over a period of two years. Residues were only detected for three (Chlormequat, Chlorothalonil and Aldicarb) of the 28 pesticides used in the experiments. As expected, residue levels were affected by the crop protection practices, but significant effects of fertility management practices were also detected. This indicates that the human health risks associated with pesticide residues may increase in low input systems which attempt to reduce the environmental impact of conventional farming systems by switching to organic matter based fertilisation regimes

Rheology at the micro-scale: new tools for bio-analysis

We present a simple and non-invasive experimental procedure to measure the linear viscoelastic properties of cells by passive particle tracking microrheology. In order to do this, a generalised Langevin equation is adopted to relate the timedependent thermal fluctuations of a probe sensor, immobilised to the cell’s membrane, to the frequency-dependent viscoelastic moduli of the cell. The method has been validated by measuring the linear viscoelastic response of a soft solid and then applied to cell physiology studies. It is shown that the viscoelastic moduli are related to the cell’s cytoskeletal structure, which in this work is modulated either by inhibiting the actin/myosin-II interactions by means of blebbistatin or by varying the solution osmolarity from iso- to hypo-osmotic conditions. The insights gained from this form of rheological analysis promises to be a valuable addition to physiological studies; e.g. cell physiology during pathology and pharmacological response

N released from organic amendments is affected by soil management history

A ryegrass bioassay was conducted to investigate the effect of soil management history on nitrogen mineralisation from composted manure and pelleted poultry manure. Soils were used from 2 field experiments comparing conventional and organic/low input management systems. When composted manure was added, soils which had received high rates of composted FYM under biodynamic management released a greater amount of nitrogen for plant uptake than those with a history of mineral or fresh manure fertilisation, suggesting that biological preconditioning may result in greater efficiency of composted FYM as a nitrogen source for plants. “Native” N mineralisation was found to be related to total soil N content