MicroBubble activated acoustic cell sorting.

Acoustophoresis, the ability to acoustically manipulate particles and cells inside a microfluidic channel, is a critical enabling technology for cell-sorting applications. However, one of the major impediments for routine use of acoustophoresis at clinical laboratory has been the reliance on the inherent physical properties of cells for separation. Here, we present a microfluidic-based microBubble-Activated Acoustic Cell Sorting (BAACS) method that rely on the specific binding of target cells to microbubbles conjugated with specific antibodies on their surface for continuous cell separation using ultrasonic standing wave. In acoustophoresis, cells being positive acoustic contrast particles migrate to pressure nodes. On the contrary, air-filled polymer-shelled microbubbles being strong negative acoustic contrast particles migrate to pressure antinodes and can be used to selectively migrate target cells. As a proof of principle, we demonstrate the separation of cancer cell line in a suspension with better than 75% efficiency. Moreover, 100% of the microbubble-cell conjugates migrated to the anti-node. Hence a better upstream affinity-capture has the potential to provide higher sorting efficiency. The BAACS technique expands the acoustic cell manipulation possibilities and offers cell-sorting solutions suited for applications at point of care.

Lab-in-a-fiber-based integrated particle separation and counting.

An all-fiber integrated device capable of separating and counting particles is presented. A sequence of silica fiber capillaries with various diameters and longitudinal cavities are used to fabricate the component for size-based elasto-inertial passive separation of particles followed by detection in an uninterrupted continuous flow. Experimentally, fluorescent particles of 1 µm and 10 µm sizes are mixed in a visco-elastic fluid and fed into the all-fiber separation component. The particles are sheathed by an elasticity enhancer (PEO - polyethylene oxide) to the side walls. Larger 10 µm particles migrate to the center of the silica capillary due to the combined inertial lift force and elastic force, while the smaller 1 µm particles are unaffected, and exit from a side capillary. A separation efficiency of 100% for the 10 µm and 97% for the 1 µm particles is achieved at a total flow rate of 50 µL min-1. To the best of our knowledge, this is the first time effective inertial-based separation has been demonstrated in circular cross-section microchannels. In the following step, the separated 10 µm particles are routed through another all-fiber component for counting and a counting throughput of ∼1400 particles per min is demonstrated. We anticipate the ability to combine high throughput separation and precise 3D control of particle position for ease of counting will aid in the development of advanced microflow cytometers capable of particle separation and quantification for various biomedical applications.

Microfluidic centrifugation assisted precipitation based DNA quantification.

Nucleic acid amplification methods are increasingly being used to detect trace quantities of DNA in samples for various diagnostic applications. However, quantifying the amount of DNA from such methods often requires time consuming purification, washing or labeling steps. Here, we report a novel microfluidic centrifugation assisted precipitation (µCAP) method for single-step DNA quantification. The method is based on formation of a visible precipitate, which can be quantified, when an intercalating dye (GelRed) is added to the DNA sample and centrifuged for a few seconds. We describe the mechanism leading to the precipitation phenomenon. We utilize centrifugal microfluidics to precisely control the formation of the visible and quantifiable mass. Using a standard CMOS sensor for imaging, we report a detection limit of 45 ng µl-1. Furthermore, using an integrated lab-on-DVD platform we recently developed, the detection limit is lowered to 10 ng µl-1, which is comparable to those of current commercially available instruments for DNA quantification. As a proof of principle, we demonstrate the quantification of LAMP products for a HIV-1B type genome containing plasmid on the lab-on-DVD platform. The simple DNA quantification system could facilitate advanced point of care molecular diagnostics.

High performance micro-flow cytometer based on optical fibres.

Flow cytometry is currently the gold standard for analysis of cells in the medical laboratory and biomedical research. Fuelled by the need of point-of-care diagnosis, a significant effort has been made to miniaturize and reduce cost of flow cytometers. However, despite recent advances, current microsystems remain less versatile and much slower than their large-scale counterparts. In this work, an all-silica fibre microflow cytometer is presented that measures fluorescence and scattering from particles and cells. It integrates cell transport in circular capillaries and light delivery by optical fibres. Single-stream cell focusing is performed by Elasto-inertial microfluidics to guarantee accurate and sensitive detection. The capability of this technique is extended to high flow rates (up to 800 µl/min), enabling a throughput of 2500 particles/s. The robust, portable and low-cost system described here could be the basis for a point-of-care flow cytometer with a performance comparable to commercial systems.

Microfluidic and transducer technologies for lab on a chip applications.

Point-of-care diagnostic devices typically require six distinct qualities: they must deliver at least the same sensitivity and selectivity, and for a cost per assay no greater than that of today's central lab technologies, deliver results in a short period of time (〈15 min at GP; 〈2h in hospital), be portable or at least small in scale, and require no or extremely little sample preparation. State-of-the-art devices deliver information of several markers in the same measurement.