Capillary-based multiplexed isothermal nucleic acid-based test for sexually transmitted diseases in patients

We demonstrate a multiplexed loop mediated isothermal amplification (LAMP) assay for infectious disease diagnostics, where the analytical process flow of target pathogens genomic DNA is performed manually by moving magnetic beads through a series of plugs in a capillary. Heat is provided by a water bath and the results read by the naked eye, enabling applications in low resource settings

Frequency dependence of microflows upon acoustic interactions with fluids

Rayleigh surface acoustic waves (SAWs), generated on piezoelectric substrates, can interact with liquids to generate fast streaming flows. Although studied extensively, mainly phenomenologically, the effect of the SAW frequency on streaming in fluids in constrained volumes is not fully understood, resulting in sub-optimal correlations between models and experimental observations. Using microfluidic structures to reproducibly define the fluid volume, we use recent advances modeling the body force generated by SAWs to develop a deeper understanding of the effect of acoustic frequency on the magnitude of streaming flows. We implement this as a new predictive tool using a finite element model of fluid motion to establish optimized conditions for streaming. The model is corroborated experimentally over a range of different acoustic excitation frequencies enabling us to validate a design tool, linking microfluidic channel dimensions with frequencies and streaming efficiencies. We show that in typical microfluidic chambers, the length and height of the chamber are critical in determining the optimum frequency, with smaller geometries requiring higher frequencies

Experimental and Numerical Studies on a Centrifugal Pump with 2D-Curved Blades in Cavitating Condition

In the presented study a special test-pump with 2D curvature blade geometry in cavitating and non-cavitating conditions was investigated using different experimental techniques and a 3D numerical model of cavitating flows. Experimental and numerical results concerning pump characteristics and performance breakdown were compared at different flow conditions. Appearing types of cavitation and the spatial distribution of vapour structures within the runner were also analysed

Monitoring genetic population biomarkers for wastewater-based epidemiology

We report a rapid “sample-to-answer” platform that can be used for the quantitative monitoring of genetic biomarkers within communities through the analysis of wastewater. The assay is based on the loop-mediated isothermal amplification (LAMP) of nucleic acid biomarkers and shows for the first time the ability to rapidly quantify human-specific mitochondrial DNA (mtDNA) from raw untreated wastewater samples. mtDNA provides a model population biomarker associated with carcinogenesis including breast, renal and gastric cancers. To enable a sample-to-answer, field-based technology, we integrated a filter to remove solid impurities and perform DNA extraction and enrichment into a low cost lateral flow-based test. We demonstrated mtDNA detection over seven consecutive days, achieving a limit of detection of 40 copies of human genomic DNA per reaction volume. The assay can be performed at the site of sample collection, with minimal user intervention, yielding results within 45 min and providing a method to monitor public health from wastewater

Spatially selecting single cell for lysis using light induced electric fields

An optoelectronic tweezing (OET) device, within an integrated microfluidic channel, is used to precisely select single cells for lysis among dense populations. Cells to be lysed are exposed to higher electrical fields than their neighbours by illuminating a photoconductive film underneath them. Using beam spot sizes as low as 2.5 μm, 100% lysis efficiency is reached in <1 min allowing the targeted lysis of cells

Green-function Method for Nonlinear Interactions of Elastic Waves

In the linear wave propagation regime, an analytical mesh-free Green-function decomposition has been shown as a viable alternative to FDTD and FEM. However, its expansion into nonlinear regimes has remained elusive due to the inherent linear properties of the Green-function approach. This work presents a novel frequency-domain Green function method to describe and model nonlinear wave interactions in isotropic hyperelastic media. As an example of the capabilities of the method, we detail the generation of sum frequency waves when initial quasimonochromatic waves are emitted in a fluid by finite sources. The method is supported by both numerical and experimental results using immersion ultrasonic techniques

Cycling of rational hybridization chain reaction to enable enzyme-free DNA-based clinical diagnosis

In order to combat the growing threat of global infectious diseases, there is the need for rapid diagnostic technologies that are sensitive and that can provide species specific information (as might be needed to direct therapy as resistant strains of microbes emerge). Here, we present a convenient, enzyme-free amplification mechanism for a rational hybridization chain reaction (HCR), which is implemented in a simple format for isothermal amplification and sensing, applied to the DNA-based diagnosis of Hepatitis B virus (HBV) in 54 patients. During the cycled amplification process, DNA monomers self-assemble in an organized and controllable way, only when a specific target HBV sequence is present. This mechanism is confirmed using super-resolution Stochastic Optical Reconstruction Microscopy (STORM). The enabled format is designed in a manner analogous to an enzyme-linked immunosorbent assay (ELISA), generating coloured products with distinct tonality and with a limit of detection of ca. 5 copies/reaction. This routine assay also showed excellent sensitivity (>97%) in clinical samples demonstrating the potential of this convenient, low cost, enzyme-free method for use in low resource settings

Visual detection of Brucella in bovine biological samples using DNA-activated gold nanoparticles

Brucellosis is a bacterial disease, which, although affecting cattle primarily, has been associated with human infections, making its detection an important challenge. The existing gold standard diagnosis relies on the culture of bacteria which is a lengthy and costly process, taking up to 45 days. New technologies based on molecular diagnosis have been proposed, either through dip-stick, immunological assays, which have limited specificity, or using nucleic acid tests, which enable to identify the pathogen, but are impractical for use in the field, where most of the reservoir cases are located. Here we demonstrate a new test based on hybridization assays with metal nanoparticles, which, upon detection of a specific pathogen-derived DNA sequence, yield a visual colour change. We characterise the components used in the assay with a range of analytical techniques and show sensitivities down to 1000 cfu/ml for the detection of Brucella. Finally, we demonstrate that the assay works in a range of bovine samples including semen, milk and urine, opening up the potential for its use in the field, in low-resource settings

Frequency dependence of microflows upon acoustic interactions with fluids

Rayleigh surface acoustic waves (SAWs), generated on piezoelectric substrates, can interact with liquids to generate fast streaming flows. Although studied extensively, mainly phenomenologically, the effect of the SAW frequency on streaming in fluids in constrained volumes is not fully understood, resulting in sub-optimal correlations between models and experimental observations. Using microfluidic structures to reproducibly define the fluid volume, we use recent advances modeling the body force generated by SAWs to develop a deeper understanding of the effect of acoustic frequency on the magnitude of streaming flows. We implement this as a new predictive tool using a finite element model of fluid motion to establish optimized conditions for streaming. The model is corroborated experimentally over a range of different acoustic excitation frequencies enabling us to validate a design tool, linking microfluidic channel dimensions with frequencies and streaming efficiencies. We show that in typical microfluidic chambers, the length and height of the chamber are critical in determining the optimum frequency, with smaller geometries requiring higher frequencies

Channel integrated optoelectronic tweezer chip for microfluidic particle manipulation

Light patterned electrical fields have been widely used for the manipulation of microparticles, from cells to microscopic electronic components. In this work, we explore a novel electromechanical phenomenon for particle focusing and sorting where the electrical field patterns are shaped by a combination of the light patterned photoconductor and the channel geometry. This effect results from the combination of particle polarisation described by the Clausius–Mossotti relation and the engineering of large electric gradients produced by choosing the channels height to suit the size of the particles being manipulated. The matched geometry increases the distortion of the field created by a combination of the illuminated photoconductor and the particles themselves and hence the non-uniformity of the field they experience. We demonstrate a new channel integration strategy which allows the creation of precisely defined channel structures in the OET device. By defining channels in photoresist sandwiched between upper and lower ITO coated glass substrates we produce robust channels of well controlled height tailored to the particle. Uniquely, the top substrate is attached before photolithographically defining the channels. We demonstrate versatile control using this effect with dynamically reconfigurable light patterns allowing the retention against flow, focusing and sorting of micro particles within the channels. Contrary to traditional designs, this channel integrated device allows patterned micro channels to be used in conjunction with conductive top and bottom electrodes producing optimal conditions for the dielectrophoretic manipulation as demonstrated by the rapid flow (up to 5 mm s−1) in which the particles can be focused