Large-Area Resistive Strain Sensing Sheet for Structural Health Monitoring.

Damage significantly influences response of a strain sensor only if it occurs in the proximity of the sensor. Thus, two-dimensional (2D) sensing sheets covering large areas offer reliable early-stage damage detection for structural health monitoring (SHM) applications. This paper presents a scalable sensing sheet design consisting of a dense array of thin-film resistive strain sensors. The sensing sheet is fabricated using flexible printed circuit board (Flex-PCB) manufacturing process which enables low-cost and high-volume sensors that can cover large areas. The lab tests on an aluminum beam showed the sheet has a gauge factor of 2.1 and has a low drift of 1.5 µ ϵ / d a y . The field test on a pedestrian bridge showed the sheet is sensitive enough to track strain induced by the bridge's temperature variations. The strain measured by the sheet had a root-mean-square (RMS) error of 7 µ ϵ r m s compared to a reference strain on the surface, extrapolated from fiber-optic sensors embedded within the bridge structure. The field tests on an existing crack showed that the sensing sheet can track the early-stage damage growth, where it sensed 600 µ ϵ peak strain, whereas the nearby sensors on a damage-free surface did not observe significant strain change.

Ancient hot and cold genes and chemotherapy resistance emergence.

We use a microfabricated ecology with a doxorubicin gradient and population fragmentation to produce a strong Darwinian selective pressure that drives forward the rapid emergence of doxorubicin resistance in multiple myeloma (MM) cancer cells. RNA sequencing of the resistant cells was used to examine (i) emergence of genes with high de novo substitution densities (i.e., hot genes) and (ii) genes never substituted (i.e., cold genes). The set of cold genes, which were 21% of the genes sequenced, were further winnowed down by examining excess expression levels. Both the most highly substituted genes and the most highly expressed never-substituted genes were biased in age toward the most ancient of genes. This would support the model that cancer represents a revision back to ancient forms of life adapted to high fitness under extreme stress, and suggests that these ancient genes may be targets for cancer therapy.

Strain Transfer for Optimal Performance of Sensing Sheet.

Sensing sheets based on Large Area Electronics (LAE) and Integrated Circuits (ICs) are novel sensors designed to enable reliable early-stage detection of local unusual structural behaviors. Such a device consists of a dense array of strain sensors, patterned onto a flexible polyimide substrate along with associated electronics. Previous tests performed on steel specimens equipped with sensing sheet prototypes and subjected to fatigue cracking pointed to a potential issue: individual sensors that were on or near a crack would immediately be damaged by the crack, thereby rendering them useless in assessing the size of the crack opening or to monitor future crack growth. In these tests, a stiff adhesive was used to bond the sensing sheet prototype to the steel specimen. Such an adhesive provided excellent strain transfer, but it also caused premature failure of individual sensors within the sheet. Therefore, the aim of this paper is to identify an alternative adhesive that survives minor damage, yet provides strain transfer that is sufficient for reliable early-stage crack detection. A sensor sheet prototype is then calibrated for use with the selected adhesive.

Automated leukocyte processing by microfluidic deterministic lateral displacement.

We previously developed a Deterministic Lateral Displacement (DLD) microfluidic method in silicon to separate cells of various sizes from blood (Davis et al., Proc Natl Acad Sci 2006;103:14779-14784; Huang et al., Science 2004;304:987-990). Here, we present the reduction-to-practice of this technology with a commercially produced, high precision plastic microfluidic chip-based device designed for automated preparation of human leukocytes (white blood cells; WBCs) for flow cytometry, without centrifugation or manual handling of samples. After a human blood sample was incubated with fluorochrome-conjugated monoclonal antibodies (mAbs), the mixture was input to a DLD microfluidic chip (microchip) where it was driven through a micropost array designed to deflect WBCs via DLD on the basis of cell size from the Input flow stream into a buffer stream, thus separating WBCs and any larger cells from smaller cells and particles and washing them simultaneously. We developed a microfluidic cell processing protocol that recovered 88% (average) of input WBCs and removed 99.985% (average) of Input erythrocytes (red blood cells) and >99% of unbound mAb in 18 min (average). Flow cytometric evaluation of the microchip Product, with no further processing, lysis or centrifugation, revealed excellent forward and side light scattering and fluorescence characteristics of immunolabeled WBCs. These results indicate that cost-effective plastic DLD microchips can speed and automate leukocyte processing for high quality flow cytometry analysis, and suggest their utility for multiple other research and clinical applications involving enrichment or depletion of common or rare cell types from blood or tissue samples. © 2016 International Society for Advancement of Cytometry.

Cell motility and drug gradients in the emergence of resistance to chemotherapy.

The emergence of resistance to chemotherapy by cancer cells, when combined with metastasis, is the primary driver of mortality in cancer and has proven to be refractory to many efforts. Theory and computer modeling suggest that the rate of emergence of resistance is driven by the strong selective pressure of mutagenic chemotherapy and enhanced by the motility of mutant cells in a chemotherapy gradient to areas of higher drug concentration and lower population competition. To test these models, we constructed a synthetic microecology which superposed a mutagenic doxorubicin gradient across a population of motile, metastatic breast cancer cells (MDA-MB-231). We observed the emergence of MDA-MB-231 cancer cells capable of proliferation at 200 nM doxorubicin in this complex microecology. Individual cell tracking showed both movement of the MDA-MB-231 cancer cells toward higher drug concentrations and proliferation of the cells at the highest doxorubicin concentrations within 72 h, showing the importance of both motility and drug gradients in the emergence of resistance.

Minimization of thermodynamic costs in cancer cell invasion.

Metastasis, the truly lethal aspect of cancer, occurs when metastatic cancer cells in a tumor break through the basement membrane and penetrate the extracellular matrix. We show that MDA-MB-231 metastatic breast cancer cells cooperatively invade a 3D collagen matrix while following a glucose gradient. The invasion front of the cells is a dynamic one, with different cells assuming the lead on a time scale of 70 h. The front cell leadership is dynamic presumably because of metabolic costs associated with a long-range strain field that precedes the invading cell front, which we have imaged using confocal imaging and marker beads imbedded in the collagen matrix. We suggest this could be a quantitative assay for an invasive phenotype tracking a glucose gradient and show that the invading cells act in a cooperative manner by exchanging leaders in the invading front.

Low-Temperature Synthesis of a TiO2/Si Heterojunction.

The classical SiO2/Si interface, which is the basis of integrated circuit technology, is prepared by thermal oxidation followed by high temperature (>800 °C) annealing. Here we show that an interface synthesized between titanium dioxide (TiO2) and hydrogen-terminated silicon (H:Si) is a highly efficient solar cell heterojunction that can be prepared under typical laboratory conditions from a simple organometallic precursor. A thin film of TiO2 is grown on the surface of H:Si through a sequence of vapor deposition of titanium tetra(tert-butoxide) (1) and heating to 100 °C. The TiO2 film serves as a hole-blocking layer in a TiO2/Si heterojunction solar cell. Further heating to 250 °C and then treating with a dilute solution of 1 yields a hole surface recombination velocity of 16 cm/s, which is comparable to the best values reported for the classical SiO2/Si interface. The outstanding performance of this heterojunction is attributed to Si-O-Ti bonding at the TiO2/Si interface, which was probed by angle-resolved X-ray photoelectron spectroscopy. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) showed that Si-H bonds remain even after annealing at 250 °C. The ease and scalability of the synthetic route employed and the quality of the interface it provides suggest that this surface chemistry has the potential to enable fundamentally new, efficient silicon solar cell devices.

Concentrating genomic length DNA in a microfabricated array.

We demonstrate that a microfabricated bump array can concentrate genomic-length DNA molecules efficiently at continuous, high flow velocities, up to 40 µm/s, if the single-molecule DNA globule has a sufficiently large shear modulus. Increase in the shear modulus is accomplished by compacting the DNA molecules to minimal coil size using polyethylene glycol (PEG) derived depletion forces. We map out the sweet spot, where concentration occurs, as a function of PEG concentration and flow speed using a combination of theoretical analysis and experiment. Purification of DNA from enzymatic reactions for next-generation DNA-sequencing libraries will be an important application of this development.

Hydrodynamic metamaterials: microfabricated arrays to steer, refract, and focus streams of biomaterials.

We show that it is possible to direct particles entrained in a fluid along trajectories much like rays of light in classical optics. A microstructured, asymmetric post array forms the core hydrodynamic element and is used as a building block to construct microfluidic metamaterials and to demonstrate refractive, focusing, and dispersive pathways for flowing beads and cells. The core element is based on the concept of deterministic lateral displacement where particles choose different paths through the asymmetric array based on their size: Particles larger than a critical size are displaced laterally at each row by a post and move along the asymmetric axis at an angle to the flow, while smaller particles move along streamline paths. We create compound elements with complex particle handling modes by tiling this core element using multiple transformation operations; we show that particle trajectories can be bent at an interface between two elements and that particles can be focused into hydrodynamic jets by using a single inlet port. Although particles propagate through these elements in a way that strongly resembles light rays propagating through optical elements, there are unique differences in the paths of our particles as compared with photons. The unusual aspects of these modular, microfluidic metamaterials form a rich design toolkit for mixing, separating, and analyzing cells and functional beads on-chip.

A microfluidic device for continuous cancer cell culture and passage with hydrodynamic forces.

We demonstrate a novel and robust microfluidic chip with combined functions of continuous culture and output of PC-3 prostate cancer cells. With digital controls, polydimethylsiloxane (PDMS) flexible diaphragms are able to apply hydrodynamic shear forces on cultures, detaching a fraction of attached cancer cells from the surface for output while leaving others for reuse in subsequent cultures. The fractions of detached cells and remaining cells can be precisely controlled. The system has not only the advantages of small size, high cell culture efficiency, and digital control, but also of simple fabrication at low cost, easy operation and robust performance. The chip performs 9 passages during 30 days of continuous culture and shows promise as a durable design suitable for long-term cell output.

Scaling of deterministic lateral displacement devices to a single column of bumping obstacles.

We describe a deterministic lateral displacement (DLD) for particle separation with only a single column of bumping features. The bifurcation of fluid streams at obstacles is not set by the "tilt" of columns with respect to macroscopic current flow, but rather by the fluidic resistances for lateral flow at each obstacle. With one column of 14 bumping features and corresponding inlet/outlet channels, the single-column DLD can separate particles with diameters of 4.8 µm and 9.9 µm at 30 µL min-1, with an area of only 0.37 mm × 1.5 mm (0.55 mm2). The large-cell output contains over 99% of the 9.9 µm particles and only 0.2% of the 4.8 m particles. The throughput per area of 54 µL min-1 per mm2 represents a 10× increase over previous selective harvesting reports for microfluidic devices in a similar particle size range.

An in vitro tumor swamp model of heterogeneous cellular and chemotherapeutic landscapes.

The heterogenous, highly metabolic stressed, poorly irrigated, solid tumor microenvironment - the tumor swamp - is widely recognized to play an important role in cancer progression as well as the development of therapeutic resistance. It is thus important to create realistic in vitro models within the therapeutic pipeline that can recapitulate the fundamental stress features of the tumor swamp. Here we describe a microfluidic system which generates a chemical gradient within connected microenvironments achieved through a static diffusion mechanism rather than active pumping. We show that the gradient can be stably maintained for over a week. Due to the accessibility and simplicity of the experimental platform, the system allows for not only well-controlled continuous studies of the interactions among various cell types at single-cell resolution, but also parallel experimentation for time-resolved downstream cellular assays on the time scale of weeks. This approach enables simple, compact implementation and is compatible with existing 6-well imaging technology for simultaneous experiments. As a proof-of-concept, we report the co-culture of a human bone marrow stromal cell line and a bone-metastatic prostate cancer cell line using the presented device, revealing on the same chip a transition in cancer cell survival as a function of drug concentration on the population level while exhibiting an enrichment of poly-aneuploid cancer cells (PACCs) as an evolutionary consequence of high stress. The device allows for the quantitative study of cancer cell dynamics on a stress landscape by real-time monitoring of various cell types with considerable experimental throughput.

The role of heterogeneous environment and docetaxel gradient in the emergence of polyploid, mesenchymal and resistant prostate cancer cells.

The ability of a population of PC3 prostate epithelial cancer cells to become resistant to docetaxel therapy and progress to a mesenchymal state remains a fundamental problem. The progression towards resistance is difficult to directly study in heterogeneous ecological environments such as tumors. In this work, we use a micro-fabricated "evolution accelerator" environment to create a complex heterogeneous yet controllable in-vitro environment with a spatially-varying drug concentration. With such a structure we observe the rapid emergence of a surprisingly large number of polyploid giant cancer cells (PGCCs) in regions of very high drug concentration, which does not occur in conventional cell culture of uniform concentration. This emergence of PGCCs in a high drug environment is due to migration of diploid epithelial cells from regions of low drug concentration, where they proliferate, to regions of high drug concentration, where they rapidly convert to PGCCs. Such a mechanism can only occur in spatially-varying rather than homogeneous environments. Further, PGCCs exhibit increased expression of the mesenchymal marker ZEB1 in the same high-drug regions where they are formed, suggesting the possible induction of an epithelial to mesenchymal transition (EMT) in these cells. This is consistent with prior work suggesting the PGCC cells are mediators of resistance in response to chemotherapeutic stress. Taken together, this work shows the key role of spatial heterogeneity and the migration of proliferative diploid cells to form PGCCs as a survival strategy for the cancer population, with implications for new therapies.

Generation of Heterogeneous Drug Gradients Across Cancer Populations on a Microfluidic Evolution Accelerator for Real-Time Observation.

Conventional cell culture remains the most frequently used preclinical model, despite its proven limited ability to predict clinical results in cancer. Microfluidic cancer-on-chip models have been proposed to bridge the gap between the oversimplified conventional 2D cultures and more complicated animal models, which have limited ability to produce reliable and reproducible quantitative results. Here, we present a microfluidic cancer-on-chip model that reproduces key components of a complex tumor microenvironment in a comprehensive manner, yet is simple enough to provide robust quantitative descriptions of cancer dynamics. This microfluidic cancer-on-chip model, the "Evolution Accelerator," breaks down a large population of cancer cells into an interconnected array of tumor microenvironments while generating a heterogeneous chemotherapeutic stress landscape. The progression and the evolutionary dynamics of cancer in response to drug gradient can be monitored for weeks in real time, and numerous downstream experiments can be performed complementary to the time-lapse images taken through the course of the experiments.

Hybrid LAE-CMOS Force-Sensing System Employing TFT-Based Compressed Sensing for Scalability of Tactile Sensing Skins.

Tactile sensing requires form-fitting and dense sensor arrays over large-areas. Hybrid systems, combining Large-Area Electronics (LAE) and silicon-CMOS ICs to respectively provide diverse sensing and high-performance computation/control, enable a platform for such sensing. A key challenge is that hybrid systems require a large number of interfaces between the LAE and CMOS domains, particularly as the number of sensors scales. This paper presents an architecture that exploits the attribute of signal sparsity, commonly exhibited in large-scale tactile-sensing applications, to reduce the interfacing complexity to a level set by the sparsity rather than the number of sensors. This enhances scalability compared to sequential-scanning and active-matrix approaches. The architecture implements compressed sensing via thin-film-transistor (TFT) switches, and is demonstrated in a force-sensing system with 20 force sensors, a TFT die (with 161 ZnO TFTs) per sensor, and a custom CMOS IC for system readout and control. Acquisition error of 0.7 k[Formula: see text] is achieved over a 100 k Ω-20 k Ω sensing range, at energy and rate of 2.46  µ J/frame and 31 fps.

Crossing microfluidic streamlines to lyse, label and wash cells.

We present a versatile method for continuous-flow, on-chip biological processing of cells, large bio-particles, and functional beads. Using an asymmetric post array in pressure-driven microfluidic flow, we can move particles of interest across multiple, independent chemical streams, enabling sequential chemical operations. With this method, we demonstrate on-chip cell treatments such as labeling and washing, and bacterial lysis and chromosomal extraction. The washing capabilities of this method are particularly valuable because they allow many analytical or treatment procedures to be cascaded on a single device while still effectively isolating their reagents from cross-contamination.

Microfluidic device for label-free measurement of platelet activation.

In this work we demonstrate a new microfluidic method for the rapid assessment of platelet size and morphology in whole blood. The device continuously fractionates particles according to size by displacing them perpendicularly to the fluid flow direction in a micro-fabricated post array. Whole blood, labeled with the fluorescent, platelet specific, antibody PE-anti-CD41, was run through the device and the positions of fluorescent objects noted as they exited the array. From this, histograms of platelet size were created which show marked increases in size after exposure to thrombin or a temperature of 4 degrees C. We infer that the well known morphological changes that occur during activation are causing the observed increase in size.

Determining blood cell size using microfluidic hydrodynamics.

Microfluidic flow cytometers currently analyze far fewer parameters than conventional flow cytometry or fluorescence activated cell sorting (FACS) in order to minimize cost and complexity. There is a need for microfluidic devices that analyze more and or new cell parameters with compact and minimal means. Here we show a new and explicitly microfluidic parameter, "hydrodynamic" cell size, and compare it to forward scatter in conventional flow cytometry. The hydrodynamic size of cells is determined by the degree of lateral displacement experienced while traveling through a 1.2-mm-wide non-clogging array of micro-fabricated obstacles. We show comparable size resolution between the microfluidic device and forward scatter in conventional flow cytometry and without the need to lyse red blood cells. We use the device to differentiate healthy lymphocytes from malignant lymphocytes by size alone and we use the device to detect increased numbers of activated lymphocytes in blood as a result of exposure to staphylococcal enterotoxin B (SEB), a potential bioterror agent. Together the results demonstrate a microfluidic device that performs some of the measurement and separation tasks of a flow cytometer but at a potentially lower cost and complexity.

Deterministic Lateral Displacement: The Next-Generation CAR T-Cell Processing?

Reliable cell recovery and expansion are fundamental to the successful scale-up of chimeric antigen receptor (CAR) T cells or any therapeutic cell-manufacturing process. Here, we extend our previous work in whole blood by manufacturing a highly parallel deterministic lateral displacement (DLD) device incorporating diamond microposts and moving into processing, for the first time, apheresis blood products. This study demonstrates key metrics of cell recovery (80%) and platelet depletion (87%), and it shows that DLD T-cell preparations have high conversion to the T-central memory phenotype and expand well in culture, resulting in twofold greater central memory cells compared to Ficoll-Hypaque (Ficoll) and direct magnetic approaches. In addition, all samples processed by DLD converted to a majority T-central memory phenotype and did so with less variation, in stark contrast to Ficoll and direct magnetic prepared samples, which had partial conversion among all donors (<50%). This initial comparison of T-cell function infers that cells prepared via DLD may have a desirable bias, generating significant potential benefits for downstream cell processing. DLD processing provides a path to develop a simple closed system that can be automated while simultaneously addressing multiple steps when there is potential for human error, microbial contamination, and other current technical challenges associated with the manufacture of therapeutic cells.

Cancer dormancy and criticality from a game theory perspective.

BACKGROUND: The physics of cancer dormancy, the time between initial cancer treatment and re-emergence after a protracted period, is a puzzle. Cancer cells interact with host cells via complex, non-linear population dynamics, which can lead to very non-intuitive but perhaps deterministic and understandable progression dynamics of cancer and dormancy. RESULTS: We explore here the dynamics of host-cancer cell populations in the presence of (1) payoffs gradients and (2) perturbations due to cell migration. CONCLUSIONS: We determine to what extent the time-dependence of the populations can be quantitively understood in spite of the underlying complexity of the individual agents and model the phenomena of dormancy.