Calibration of triaxial accelerometers by constant rotation rate in the gravitational field.

We extend the use of the intrinsic properties calibration method for triaxial accelerometers that we reported previously from discrete angular steps to using a constant rotation rate to produce a time varying sinusoidal excitation in the earth's gravitational field. We show that this extension yields the low frequency calibration response of the device under test. Whereas traditional vibration-based methods using shakers generally exhibit an increased measurement uncertainty with decreased excitation frequency, we show that this approach does not. We report results obtained from a commercial triaxial digital accelerometer from DC up to a 0.5 Hz rotation rate. The maximum rotation rate that we report is limited by our rotation stage; but we expect that the method can be extended to higher rotation rates with an upper limit constrained by what can be tolerated as a maximum centripetal acceleration.

Reduction of calibration uncertainty due to mounting of three-axis accelerometers using the intrinsic properties model.

We show that the calibration of tri-axis accelerometers based on the device's intrinsic properties alleviates the uncertainty due to mounting misalignment in comparison to the use of the sensitivity matrix. The intrinsic properties of a tri-axis accelerometer are based on a (u, v, w) coordinate system that represent the direction of maximum sensitivities of each of the three accelerometers (U, V, W) and are assumed not to be perfectly orthogonal to each other. The calibration procedure requires rotation of the device in the gravitational field around each of the Cartesian coordinate (x, y, z) axes. One component in driving down the uncertainty of laboratory comparisons and calibration repeats relates to misalignment in mounting the device onto the calibration instrument. We show that the uncertainty of the cross-axis terms of the sensitivity matrix is a dominating factor affecting uncertainty down to a 0.01° misalignment at a 100 µV noise level. The misalignment component can be exacerbated when calibrating modern microelectromechanical systems (MEMS)-based accelerometers, which are typically a few millimeters in dimension.

Shock Measurements Based on Pendulum Excitation and Laser Doppler Velocimetry: Primary Calibration by SI-Traceable Distance Measurements.

A new method is described to provide a primary calibration of shock measurements produced by a shock measurement system consisting of pendulum excitation and laser Doppler velocimetry. The method uses the laser Doppler velocimeter to determine the total distance traveled by a rigid block that slides along a Teflon (fluorocarbon) channel after being struck by a pendulum head, and the resulting distance is compared to the distance measured by an SI-traceable length measurement. The instantaneous velocity of the block is measured by the velocimeter and is used to calculate the displacement of the block by integrating the velocity data. The result is compared to the displacement measured using calibrated rulers and calipers. The method was applied to an independently calibrated commercial velocimeter for impact accelerations ranging from 2000 to 30,000 m/s2. The results of the independent mechanical displacement measurements agreed with those from the commercial velocimeter to within ±0.3 %, with better agreement above accelerations of order 10,000 m/s2 to within ±0.1 %. A conservative, upper-bound, uncertainty analysis included the effects of noise and other random errors, as well as type B estimates for systematic errors from occasional momentary demodulation failures (dropouts), use of a different number of rulers before and after shock distance measurement, and the relative frequency response of the velocimeter.

Gravity-Based Characterization of Three-Axis Accelerometers in Terms of Intrinsic Accelerometer Parameters.

Cross-sensitivity matrices are used to translate the response of three-axis accelerometers into components of acceleration along the axes of a specified coordinate system. For inertial three-axis accelerometers, this coordinate system is often defined by the axes of a gimbal-based instrument that exposes the device to different acceleration inputs as the gimbal is rotated in the local gravitational field. Therefore, the cross-sensitivity matrix for a given three-axis accelerometer is not unique. Instead, it depends upon the orientation of the device when mounted on the gimbal. We define nine intrinsic parameters of three-axis accelerometers and describe how to measure them directly and how to calculate them from independently determined cross-sensitivity matrices. We propose that comparisons of the intrinsic parameters of three axis accelerometers that were calculated from independently determined cross-sensitivity matrices can be useful for comparisons of the cross-sensitivity-matrix measurement capability of different institutions because the intrinsic parameters will separate the accelerator-gimbal alignment differences among the participating institutions from the purely gimbal-related differences, such as gimbal-axis orthogonality errors, z-axis gravitational-field alignment errors, and angle-setting or angle-measurement errors.

Hybrid cell adhesive material for instant dielectrophoretic cell trapping and long-term cell function assessment.

Dielectrophoresis (DEP) for cell manipulation has focused, for the most part, on approaches for separation/enrichment of cells of interest. Advancements in cell positioning and immobilization onto substrates for cell culture, either as single cells or as cell aggregates, has benefited from the intensified research efforts in DEP (electrokinetic) manipulation. However, there has yet to be a DEP approach that provides the conditions for cell manipulation while promoting cell function processes such as cell differentiation. Here we present the first demonstration of a system that combines DEP with a hybrid cell adhesive material (hCAM) to allow for cell entrapment and cell function, as demonstrated by cell differentiation into neuronlike cells (NLCs). The hCAM, comprised of polyelectrolytes and fibronectin, was engineered to function as an instantaneous cell adhesive surface after DEP manipulation and to support long-term cell function (cell proliferation, induction, and differentiation). Pluripotent P19 mouse embryonal carcinoma cells flowing within a microchannel were attracted to the DEP electrode surface and remained adhered onto the hCAM coating under a fluid flow field after the DEP forces were removed. Cells remained viable after DEP manipulation for up to 8 d, during which time the P19 cells were induced to differentiate into NLCs. This approach could have further applications in areas such as cell-cell communication, three-dimensional cell aggregates to create cell microenvironments, and cell cocultures.

Separation and metrology of nanoparticles by nanofluidic size exclusion.

A nanofluidic technology for the on-chip size separation and metrology of nanoparticles is demonstrated. A nanofluidic channel was engineered with a depth profile approximated by a staircase function. Numerous stepped reductions in channel depth were used to separate a bimodal mixture of nanoparticles by nanofluidic size exclusion. Epifluorescence microscopy was used to map the size exclusion positions of individual nanoparticles to corresponding channel depths, enabling measurement of the nanoparticle size distributions and validation of the size separation mechanism.

Quantum dot FRET-based probes in thin films grown in microfluidic channels.

This paper describes the development of new fluorescence resonance energy transfer (FRET)-based quantum dot probes for proteolytic activity. The CdSe/ZnS quantum dots are incorporated into a thin polymeric film, which is prepared by layer-by-layer deposition of alternately charged polyelectrolytes. The quantum dots, which serve as fluorescent donors, are separated from rhodamine acceptor molecules, which are covalently attached to the film surface by a varying number of polyelectrolyte layers. When excited with visible light, the emission color of the polyelectrolyte multilayer film appears orange due to FRET between the quantum dots and molecular acceptors. The emission color changes to green when the rhodamine molecules are removed from the surface by enzymatic cleavage. The new probe design enables the use of quantum dots in bioassays, in this study for real-time monitoring of trypsin activity, while alleviating concerns about their potential toxicity. Application of these quantum dot FRET-based probes in microfluidic channels enables bioanalysis of volume-limited samples and single-cell studies in an in vivo-like environment.

Accurate optical analysis of single-molecule entrapment in nanoscale vesicles.

We present a nondestructive method to accurately characterize low analyte concentrations (0-10 molecules) in nanometer-scale lipid vesicles. Our approach is based on the application of fluorescence fluctuation analysis (FFA) and multiangle laser light scattering (MALLS) in conjunction with asymmetric field flow fractionation (AFFF) to measure the entrapment efficiency (the ratio of the concentration of encapsulated dye to the initial bulk concentration) of an ensemble of liposomes with an average diameter less than 100 nm. Water-soluble sulforhodamine B (SRB) was loaded into the aqueous interior of nanoscale liposomes synthesized in a microfluidic device. A confocal microscope was used to detect a laser-induced fluorescence signal resulting from both encapsulated and unencapsulated SRB molecules. The first two cumulants of this signal along with the autocorrelation function (ACF) were used to quantify liposome entrapment efficiency. Our analysis moves beyond typical, nonphysical assumptions of equal liposome size and brightness. These advances are essential for characterizing liposomes in the single-molecule encapsulation regime. Our work has further analytical impact because it could increase the interrogation time of free-solution molecular analysis by an order of magnitude and form the basis for the development of liposome standard reference materials.

Microfluidic directed self-assembly of liposome-hydrogel hybrid nanoparticles.

We present a microfluidic method to direct the self-assembly of temperature-sensitive liposome-hydrogel hybrid nanoparticles. Our approach yields nanoparticles with structural properties and highly monodisperse size distributions precisely controlled across a broad range relevant to the targeted delivery and controlled release of encapsulated therapeutic agents. We used microfluidic hydrodynamic focusing to control the convective-diffusive mixing of two miscible nanoparticle precursor solutions (a DPPC:cholesterol:DCP phospholipid formulation in isopropanol and a photopolymerizable N-isopropylacrylamide mixture in aqueous buffer) to form nanoscale lipid vesicles with encapsulated hydrogel precursors. These precursor nanoparticles were collected off-chip and were irradiated with ultraviolet (UV) light in bulk to polymerize the nanoparticle interiors into hydrogel cores. Multiangle laser light scattering in conjunction with asymmetric flow field-flow fractionation was used to characterize nanoparticle size distributions, which spanned the approximately 150 to approximately 300 nm diameter range as controlled by microfluidic mixing conditions, with a polydispersity of approximately 3% to approximately 5% (relative standard deviation). Transmission electron microscopy was then used to confirm the spherical shape and core-shell composition of the hybrid nanoparticles. This method may be extended to the directed self-assembly of other similar cross-linked hybrid nanoparticle systems with engineered size/structure-function relationships for practical use in healthcare and life science applications.

Polyelectrolyte Multilayer-Treated Electrodes for Real-Time Electronic Sensing of Cell Proliferation.

We report on the use of polyelectrolyte multilayer (PEM) coatings as a non-biological surface preparation to facilitate uniform cell attachment and growth on patterned thin-film gold (Au) electrodes on glass for impedance-based measurements. Extracellular matrix (ECM) proteins are commonly utilized as cell adhesion promoters for electrodes; however, they exhibit degradation over time, thereby imposing limitations on the duration of conductance-based biosensor experiments. The motivation for the use of PEM coatings arises from their long-term surface stability as promoters for cell attachment, patterning, and culture. In this work, a cell proliferation monitoring device was fabricated. It consisted of thin-film Au electrodes deposited with a titanium-tungsten (TiW) adhesion layer that were patterned on a glass substrate and passivated to create active electrode areas. The electrode surfaces were then treated with a poly(ethyleneimine) (PEI) anchoring layer and subsequent bilayers of sodium poly(styrene sulfonate) (PSS) and poly(allylamine hydrochloride) (PAH). NIH-3T3 mouse embryonic fibroblast cells were cultured on the device, observed by optical microscopy, and showed uniform growth characteristics similar to those observed on a traditional polystyrene cell culture dish. The optical observations were correlated to electrical measurements on the PEM-treated electrodes, which exhibited a rise in impedance with cell proliferation and stabilized to an approximate 15 % increase as the culture approached confluency. In conclusion, cells proliferate uniformly over gold and glass PEM-treated surfaces, making them useful for continuous impedance-based, real-time monitoring of cell proliferation and for the determination of cell growth rate in cellular assays.

Generalized temperature measurement equations for Rhodamine B dye solution and its application to microfluidics.

Temperature mapping based on fluorescent signal intensity ratios is a widely used noncontact approach for investigating temperature distributions in various systems. This noninvasive method is especially useful for applications, such as microfluidics, where accurate temperature measurements are difficult with conventional physical probes. However, the application of a calibration equation to relate fluorescence intensity ratio to temperature is not straightforward when the reference temperature in a given application is different than the one used to derive the calibration equation. In this report, we develop and validate generalized calibration equations that can be applied for any value of reference temperature. Our analysis shows that a simple linear correction for a 40 degrees C reference temperature produces errors in measured temperatures between -3 to 8 degrees C for three previously published sets of cubic calibration equations. On the other hand, corrections based on an exact solution of these equations restrict the errors to those inherent in the calibration equations. The methods described here are demonstrated for cubic calibration equations derived by three different groups, but the general method can be applied to other dyes and calibration equations.

Nanofluidic structures with complex three-dimensional surfaces.

Nanofluidic devices have typically explored a design space of patterns limited by a single nanoscale structure depth. A method is presented here for fabricating nanofluidic structures with complex three-dimensional (3D) surfaces, utilizing a single layer of grayscale photolithography and standard integrated circuit manufacturing tools. This method is applied to construct nanofluidic devices with numerous (30) structure depths controlled from approximately 10 to approximately 620 nm with an average standard deviation of <10 nm over distances of >1 cm. A prototype 3D nanofluidic device is demonstrated that implements size exclusion of rigid nanoparticles and variable nanoscale confinement and deformation of biomolecules.

Single molecule measurements within individual membrane-bound ion channels using a polymer-based bilayer lipid membrane chip.

The measurement of single poly(ethylene glycol) (PEG) molecules interacting with individual bilayer lipid membrane-bound ion channels is presented. Measurements were performed within a polymer microfluidic system including an open-well bilayer lipid membrane formation site, integrated Ag/AgCl reference electrodes for on-chip electrical measurements, and multiple microchannels for independent ion channel and analyte delivery. Details of chip fabrication, bilayer membrane formation, and alpha-hemolysin ion channel incorporation are discussed, and measurements of interactions between the membrane-bound ion channels and single PEG molecules are presented.

Dielectrophoretic capture of mammalian cells using transparent indium tin oxide electrodes in microfluidic systems.

Transparent indium tin oxide microelectrodes were fabricated and used to immobilize suspended NIH 3T3 fibroblast cells by positive dielectrophoresis. The indium tin oxide electrodes facilitated microscopic observation of immobilized cells compared with opaque metallized electrodes. Dielectrophoresis was used to capture arrays of individual cells and form small cell clusters within a microfluidic network. The extent of cellular immobilization (no-cell, single-cell, or multiple-cell capture) was correlated with the applied voltage and inversely with the flow velocity. Specific conditions yielding predominantly single-cell capture were identified. The viability of immobilized cells was confirmed using fluorescence microscopy.

Microwave Power Absorption in Low-Reflectance, Complex, Lossy Transmission Lines.

Simple sets of equations have been derived to describe the absorption of microwave power in three-region, lossy transmission lines in terms of S-parameter reflection and transmission amplitudes. Each region was assumed to be homogeneous with discontinuities at the region boundaries. Different sets of equations were derived to describe different assumptions about the amplitudes of the reflection coefficients at the different boundaries. These equations, which are useful when interference effects due to multiple reflections are small, were used to analyze S-parameter measurements on a transmission line that had a microfluidic channel in its middle region. The channel was empty for one set of measurements and filled with water for a second set of measurements. Most of the reflection assumptions considered here produced similar results for the fraction of the applied microwave power that was absorbed by a water-filled microchannel. This shows that the absorbed power is relatively insensitive to the reflection details as long as energy is conserved in the analysis. Another important result of this work is that the difference between the power absorbed in a water-filled channel and the power absorbed in the same empty channel can be a poor predictor of the power absorbed in the water in the presence of competing absorption processes such as absorption by the transmission-line metal.

Fabrication of polymer microfluidic systems by hot embossing and laser ablation.

Fabrication of microfluidic channels in common commercially available thermoplastic materials can be easily accomplished using hot embossing or ultraviolet (UV) laser ablation. Hot embossing involves replication of a microfluidic network in a polymer substrate from a stamp (or template) fabricated in silicon or metal. UV laser ablation is performed by either exposing the polymer substrate through a mask or by using a laser direct-write process. The resulting polymer microfluidic channels are most often sealed with another polymer piece using thermal bonding or solvent bonding to complete the fabrication procedure. Unlike their silicon and glass counterparts, polymer microfluidic systems can be fabricated by these methods in less than 1 h, making the materials attractive for both research prototyping and commercialization.

Embedded MicroHeating Elements in Polymeric MicroChannels for Temperature Control and Fluid Flow Sensing.

This paper describes the first demonstration of temperature control and flow sensing of fluids using integrated circuit (IC)-based microheating elements embedded in microchannels molded in polydimethylsiloxane (PDMS). Fluid channels and connections to capillary tubing are molded in PDMS using a silicon wafer template. The PDMS film is then bonded to an IC that contains the micromachined microheating elements. Capillary tubes are inserted and fluids are externally pumped through the channels. Heating of the fluid is observed by the formation of bubbles on the microheating element. Sensing of fluid flow is demonstrated by measuring a change in the large signal resistance of the microheater analogous to a hot wire anemometer with a detection limit of ± 320 pL/s.

The Art in Science of MicroTAS: the 2013 edition.

The 6th annual µTAS Art in Science Award was presented to Ye Wang of Eindhoven University of Technology at the 17th International Conference of Miniaturized Systems for Chemistry and Life Sciences held in Freiburg, Germany, on October 27-31, 2013. The winning image captivated the hearts and minds of the judges and is featured on the front cover of this issue. "Artificial Life", as the author of the work that produced this image named it, was taken with a scanning electron microscope (SEM) and shows cilia-like structures (or "microhairs") generated from PDMS (polydimethylsiloxane) and magnetic nanoparticles (Y. Wang, Y. Gao, H. M. Wyss, P. D. Anderson and J. M. J. den Toonder, "Out of Cleanroom, Self-assembled Magnetic Artificial Cilia", micro-TAS, 2013, 787-789). To produce the structure the author used a glass mold produced by femtolaser modification and hydrofluoric acid etching. As the title implies, the features produced by this procedure resemble cilia-like structures as seen in a number of eukaryotic cells.

DNA molecules descending a nanofluidic staircase by entropophoresis.

A complex entropy gradient for confined DNA molecules was engineered for the first time. Following the second law of thermodynamics, this enabled the directed self-transport and self-concentration of DNA molecules. This new nanofluidic method is termed entropophoresis. As implemented in experiments, long DNA molecules were dyed with cyanine dimers, dispersed in a high ionic strength buffer, and confined by a nanofluidic channel with a depth profile approximated by a staircase function. The staircase step depths spanned the transition from strong to moderate confinement. The diffusion of DNA molecules across slitlike steps was ratcheted by entropic forces applied at step edges, so that DNA molecules descended and collected at the bottom of the staircase, as observed by fluorescence microscopy. Different DNA morphologies, lengths, and stoichiometric base pair to dye molecule ratios were tested and determined to influence the rate of transport by entropophoresis. A model of ratcheted diffusion was used to interpret a shifting balance of forces applied to linear DNA molecules of standard length in a complex free energy landscape. Related metrics for the overall and optimum performance of entropophoresis were developed. The device and method reported here transcend current limitations in nanofluidics and present new possibilities in polymer physics, biophysics, separation science, and lab-on-a-chip technology.

Microfluidic mixing and the formation of nanoscale lipid vesicles.

We investigate the formation of unilamellar lipid vesicles (liposomes) with diameters of tens of nanometers by controlled microfluidic mixing and nanoparticle determination (COMMAND). Our study includes liposome synthesis experiments and numerical modeling of our microfluidic implementation of the batch solvent injection method. We consider microfluidic liposome formation from the perspective of fluid interfaces and convective-diffusive mixing, as we find that bulk fluid flow parameters including hydrodynamically focused alcohol stream width, final alcohol concentration, and shear stress do not primarily determine the vesicle formation process. Microfluidic device geometry in conjunction with hydrodynamic flow focusing strongly influences vesicle size distributions, providing a coarse method to control liposome size, while total flow rate allows fine-tuning the vesicle size in certain focusing regimes. Although microfluidic liposome synthesis is relatively simple to implement experimentally, numerical simulations of the mixing process reveal a complex system of fluid flow and mass transfer determining the formation of nonequilibrium vesicles. These results expand our understanding of the microfluidic environment that controls liposome self-assembly and yield several technological advances for the on-chip synthesis of nanoscale lipid vesicles.