Search

Cone penetration testing (CPT) is a preferred method for characterizing soil profiles for evaluating seismic liquefaction triggering potential. However, CPT has limitations in characterizing highly stratified profiles because the measured tip resistance ( q c ) of the cone penetrometer is influenced by the properties of the soils above and below the tip. This results in measured q c values that appear “blurred” at sediment layer boundaries, inhibiting our ability to characterize thinly layered strata that are potentially liquefiable. Removing this “blur” has been previously posed as a continuous optimization problem, but in some cases this methodology has been less efficacious than desired. Thus, we propose a new approach to determine the corrected q c values (i.e. values that would be measured in a stratum absent of thin-layer effects) from measured values. This new numerical optimization algorithm searches for soil profiles with a finite number of layers which can automatically be added or removed as needed. This algorithm is provided as open-source MATLAB software. It yields corrected q c values when applied to computer-simulated and calibration chamber CPT data. We compare two versions of the new algorithm that numerically optimize different functions, one of which uses a logarithm to refine fine-scale details, but which requires longer calculation times to yield improved corrected q c profiles.

A new strategy to produce stable barcodes using biological self-assembly of streptavidin- and biotin-functionalized quantum dots is reported. Such systems are of potential use in multiplexed immunoassay and nucleic acid hybridization assays.

We report the ability to move and arrange patterns of live embryonic stem cells using holographic optical tweezers. Single cell suspensions of mouse embryonic stem cells were manipulated with holographic optical tweezers into a variety of patterns including lines, curves and circles. Individual cells were also lifted out of the sample plane highlighting the potential for 3D positional control. Trypan blue dye exclusion and Live/Dead™ staining (CMFDA−1, EthHD−1) showed that the cells were still viable after manipulation with the optical tweezers. The ability to move individual stem cells into specific, pre-defined patterns provides a method to study how arrangement and associated small-scale interactions occur between neighbouring cells.

We exploit the mechanical action of surface acoustic waves (SAW) to differentially lyse human cancer cells in a chemical-free manner. The extent to which cells were disrupted is reported for a range of SAW parameters, and we show that the presence of 10 μm polystyrene beads is required to fully rupture cells and their nuclei. We show that SAW is capable of subcellular fractionation through the chemical-free isolation of nuclei from whole cells. The concentration of protein was assessed in lysates with a sensitive microfluidic antibody capture (MAC) chip. An antibody-based sandwich assay in a microfluidic microarray format was used to detect unlabeled human tumor suppressor protein p53 in crude lysates, without any purification step, with single-molecule resolution. The results are digital, enabling sensitive quantification of proteins with a dynamic range >4 orders of magnitude. For the conditions used, the efficiency of SAW-induced mechanical lysis was determined to be 12.9% ± 0.7% of that for conventional detergent-based lysis in yielding detectable protein. A range of possible loss mechanisms that could lead to the drop in protein yield are discussed. Our results show that the methods described here are amenable to an integrated point-of-care device for the assessment of tumor protein expression in fine needle aspirate biopsies.

We have demonstrated a monolithic integrated arrayed waveguide grating (AWG) microspectrometer microfluidic platform capable of fluorescence spectroscopic analysis. The microspectrometer in this proof of concept study has a small (1 cm × 1 cm) footprint and 8 output channels centred on different wavelengths. We show that the signals from the output channels detected on a camera chip can be used to recreate the complete fluorescence spectrum of an analyte. By making fluorescence measurements of (i) mixed quantum dot solutions, (ii) an organic fluorophore (Cy5) and (iii) the propidium iodide (PI)-DNA assay, we illustrate the unique advantages of the AWG platform for simultaneous, quantitative multiplex detection and its capability to detect small spectroscopic shifts. Although the current system is designed for fluorescence spectroscopic analysis, in principle, it can be implemented for other types of analysis, such as Raman spectroscopy. Fabricated using established semiconductor industry methods, this miniaturised platform holds great potential to create a handheld, low cost biosensor with versatile detection capability.

Biosensing now spans a range of medical, environmental and industrial applications. New optical techniques based on scattering, absorbance, transmission and photonic crystal technology will allow for compact, low-cost and reliable automatic analysis that can be easily produced on a mass scale.

Holographic or diffractive optical components are widely implemented using spatial light modulators within optical tweezers to form multiple, and/or modified traps. We show that by further modifying the hologram design to account for residual aberrations, the fidelity of the focused beams can be significantly improved, quantified by a spot sharpness metric. However, the impact this improvement has on the quality of the optical trap depends upon the particle size. For particle diameters on the order of 1 microm, aberration correction can improve the trap performance metric, which is the ratio of the mean square displacement of a corrected trap to an uncorrected trap, in excess of 25%, but for larger particles the trap performance is not unduly affected by the aberrations typically encountered in commercial spatial light modulators.

A microfluidic format-based system has been developed for in situ monitoring of the calcium flux response to agonists using Chinese hamster ovary (CHO) cells. The assay is based on measuring the fluorescent intensity of the calcium-sensitive indicator, Fluo-4 AM, and was performed in a modified glass chip channel, whose surface was functionalised using a silanisation method with 3-aminopropyltriethoxysilane (APTS) (enabling the cells to be immobilised on the channel surface). CHO cells calcium flux response was measured for different agonists over a range of concentrations. Cells and reagents were introduced into the chip in a continuous flow as a series of plugs in a given sequence.

We demonstrate a technique for the multi-point measurement of fluid flow in microscopic geometries. The technique consists of an array of microprobes can be simultaneously trapped and used to map out the fluid flow in a microfluidic device. The optical traps are alternately turned on and off such that the probe particles are displaced by the flow of the surrounding fluid and then re-trapped. The particles' displacements are monitored by digital video microscopy and directly converted into velocity field values. The techniques described have potential to be extended to drive an integrated lab-on-chip device, where pumping, flow measurement and optical sensing could all be achieved by structuring a single laser beam.

Holographic or diffractive optical components, such as a spatial light modulator (SLM), can be used in optical tweezers for the creation of multiple and modified optical traps. In addition to this, SLMs can also be used to correct for aberrations within the optical train resulting in an improved trapping performance. Typically an electrically addressed SLM may deviate from flatness by up to 4λ, dominated by astigmatism due to the overall curvature of the SLM surface. This astigmatism may be corrected by adding the appropriate hologram to the SLM display resulting in a dramatic improvement in the fidelity of the focussed spot. The impact that this correction has on the performance of the optical trap is most noticeable for small particles. For the SLM used in this study, the improvement in trap performance for a 0.8 μm diameter particles can be in excess of 25%. However, for 5 μm diameter particles our results show an improvement of less than 0.5%. This dependence upon particle size is most probably associated with the relative size of the PSF and the trapped particle. Once the PSF is significantly smaller than the particle diameter, further reduction brings little improvement in trap performance.

This paper demonstrates the use of micron sized beads, modified with fluorescent dyes, as non-invasive sensors to probe the local changes in pH, within a microfluidic channel. To achieve this, amine modified polystyrene spheres (either 3 microm or 6 microm in diameter) were functionalised with the pH sensitive fluorochrome SNARF-1 to produce point sensors. The modified beads were trapped at defined positions close to a pair of integrated planar gold microelectrodes within the channel, using optical tweezers. Both transient and steady-state electrochemical potentials were applied to the microelectrode pair in order to generate changes in the local pH, associated with electrolysis. The functionalised beads indicated the pH changes in the channel, measured as a change in the fluorescence signal, generated by the immobilised pH sensitive dye. Responses were measured with temporal resolutions of between 1 and 200 ms, whilst the spatial resolution of the pH gradients was limited by the size of the beads to 3 microm.

We use holographic optical tweezers to create and monitor the liquid flow within a micro-fluidic device. Using the tweezers to both trap and spin micron-sized beads within a 10-20 micron wide channel creates a fluid flow of the order of 200 cubic microns/sec. We also use the optical tweezers to measure the fluid flow by trapping and releasing probe particles that are imaged with high temporal and spatial resolution. Using the multi-trap capability of the holographic optical tweezers we measure the transverse fluid velocity at many positions simultaneously with an accuracy of better than 1 micron/sec. Such studies are highly pertinent to lab-on-chip systems for various applications and studies within the biosciences.

Central to the success of microfluidic systems has been the development of innovative methods for the manipulation of fluids within microchannels. We demonstrate a method for generating flow within a microfluidic channel using an optically driven pump. The pump consists of two counter rotating birefringent vaterite particles trapped within a microfluidic channel and driven using optical tweezers. The transfer of spin angular momentum from a circularly polarised laser beam rotates the particles at up to 10 Hz. We show the that the pump is able to displace fluid in microchannels, with flow rates of up to 200 μm 3 s -1 (200 fL s -1 ). The direction of fluid pumping can be reversed by altering the sense of the rotation of the vaterite beads. We also incorporate a novel optical sensing method, based upon an additional probe particle, trapped within separate optical tweezers, enabling us to map the magnitude and direction of fluid flow within the channel. The techniques described in the paper have potential to be extended to drive an integrated lab-on-chip device, where pumping, flow measurement and optical sensing could all be achieved by structuring a single laser beam.

SNARF-1 fluorochrome was used to functionalize 3μm diameter latex spheres making them sensitive to the pH of their environment, manifested as a change in their fluorescence. The fluorescence emission at 580nm was excited using a filtered xenon arc lamp at 515nm. A solution of functionalized latex spheres was placed between gold microelectrodes in a microfluidic channel. Optical tweezers were used to trap and manipulate the spheres in the vicinity of the microelectrodes, to map out the pH profile in the electrolyte solution, induced by passing 20 microsecond transient current pulses through the microelectrodes.

We have developed an interactive user-interface that can be used to generate phase holograms for use with spatial light modulators. The program utilises different hologram design techniques allowing the user to select an appropriate algorithm. The program can be used to generate multiple beams, interference patterns and can be used for beam steering. We therefore see a major application of the program to be within optical tweezers to control the position, number and type of optical traps.

We use holographic optical tweezers to trap multiple micron-sized objects and manipulate them in 3-dimensions. Trapping multiple objects allow us to create 3-dimensional structures, examples of which include; simple cubes which can be rotated or scaled, complex crystal structures like the diamond lattice or interactive 3-dimensional control of trapped particles anywhere in the sample volume.

A high-density array of assay sites for cell based drug profiling was micromachined on microscope glass coverslips. The shortening and intracellular calcium of individual ventricular muscle cells electrically stimulated in picolitre volume was recorded using video enhanced light microscopy. The electromechanical coupling of cells enclosed in microchambers decayed in contrast to cells permanently superfused in microchannels, indicating ischemic conditions in the restricted volume. This cell-based model of ischemia will enable the profiling of new protective compounds in heart failure.

The micromanipulation of objects into 3-dimensional geometries within holographic optical tweezers is carried out using modified Gerchberg-Saxton (GS) and direct binary search (DBS) algorithms to produce the hologram designs. The algorithms calculate sequences of phase holograms, which are implemented using a spatial light modulator, to reconfigure the geometries of optical traps in many planes simultaneously. The GS algorithm is able to calculate holograms quickly from the initial, intermediate and final trap positions. In contrast, the DBS algorithm is slower and therefore used to pre-calculate the holograms, which are then displayed in sequence. Assembly of objects in a variety of 3-D configurations is semi-automated, once the traps in their initial positions are loaded.

We demonstrate controllable poration within ≈1 µm regions of individual cells, mediated by a near-IR laser interacting with thin-layer amorphous silicon substrates. This technique will allow new experiments in single-cell biology, particularly in neuroscience. As our understanding of the fundamental mechanistic processes underpinning biology expands, so does the need for high-precision tools to allow the dissection of the heterogeneity and stochastic processes that dominate at the single- and sub-cellular level. Here, we demonstrate a highly controllable and reproducible optical technique for inducing poration within specific regions of a target cell’s plasma membrane, permitting localized delivery of payloads, depolarization and lysis experiments to be conducted in unprecedented detail. Experiments support a novel mechanism for the process, based upon a thermally-induced change triggered by the interactions of a near-IR laser with a biocompatible thin film substrate at powers substantially below that used in standard optoporation experiments.

We have developed holographic optical tweezers that can manipulate many particles simultaneously in three dimensions in order to create micro-crystal structures that extend over many tens of microns. The technique uses specific hologram-design algorithms to create structures that can be dynamically scaled or rotated about arbitrary axes. We believe the generation and control of pre-determined crystal-like structures have significant potential in fields as diverse as photonic-crystal construction, seeding of biological tissue growth and creation of metrological standards within nanotechnology.

Silica particles were partially coated with silver, and a suitable chromophore, such that they could be trapped within an optical tweezers system and simultaneously be excited to emit a surface–enhanced resonance Raman scattering (SERRS) spectrum. A standard 1064 nm TEM00 mode laser was used to trap the bead whilst a frequency doubling crystal inserted into the beam gave several microwatts of 532 nm co-linear light to excite the SERRS emission. The configuration has clear applications in providing apparatus that can manipulate a particle whilst simultaneously obtaining surface sensitive sensory information.

We have developed software with an interactive user interface that can be used to generate phase holograms for use with spatial light modulators. The program utilizes different hologram design techniques, allowing the user to select an appropriate algorithm. The program can be used to generate multiple beams and can be used for beam steering. We see a major application of the program to be in optical tweezers to control the position, number, and type of optical traps.