Gravimetric deposition of microliter drops with radiometric confirmation.

A manual microliter gravimetric dispensing technique is demonstrated using a micropipettor modified for use with removeable microcapillaries. Liquid scintillation sources were prepared from a well-characterized 241Am reference solution, providing a radiometric check of dispensed masses. Further experiments confirmed controlled dispensing of drops onto gold foils with losses ≤0.34(4) % of the total drop activity. A detailed measurement equation for the weighing technique, including the corrections for evaporation, is presented with a full accounting of associated uncertainties.

Design of an electrostatic balance mechanism to measure optical power of 100 kW.

A new instrument is required to accommodate the need for increased portability and accuracy in laser power measurement above 100 W. Reflection and absorption of laser light provide a measurable force from photon momentum exchange that is directly proportional to laser power, which can be measured with an electrostatic balance traceable to the SI. We aim for a relative uncertainty of 10-3 with coverage factor k = 2. For this purpose, we have designed a monolithic parallelogram 4-bar linkage incorporating elastic circular notch flexure hinges. The design is optimized to address the main factors driving force measurement uncertainty from the balance mechanism: corner loading errors, balance stiffness, stress in the flexure hinges, sensitivity to vibration, and sensitivity to thermal gradients. Parasitic rotations in the free end of the 4-bar linkage during arcuate motion are constrained by machining tolerances. An analytical model shows this affects the force measurement less than 0.01 percent. Incorporating an inverted pendulum reduces the stiffness of the system without unduly increasing tilt sensitivity. Finite element modeling of the flexures is used to determine the hinge orientation that minimizes stress which is therefore expected to minimize hysteresis. Thermal effects are mitigated using an external enclosure to minimize temperature gradients, although a quantitative analysis of this effect is not carried out. These analyses show the optimized mechanism is expected to contribute less than 1 × 10-3 relative uncertainty in the final laser power measurement.

Toward a New Primary Standardization of Radionuclide Massic Activity Using Microcalorimetry and Quantitative Milligram-Scale Samples.

We present a new paradigm for the primary standardization of radionuclide activity per mass of solution (Bq/g). Two key enabling capabilities are 4π decay-energy spectrometry using chip-scale sub-Kelvin microcalorimeters and direct realization of mass by gravimetric inkjet dispensing using an electrostatic force balance. In contrast to traditional traceability, which typically relies on chemical separation of single-radionuclide samples, 4π integral counting, and additional spectrometry methods to verify purity, the system described here has both 4π counting efficiency and spectroscopic resolution sufficient to identify multiple radionuclides in the same sample at once. This enables primary standardization of activity concentrations of mixed-radionuclide samples. A major benefit of this capability, beyond metrology, is in assay of environmental and forensics samples, for which the quantification of multiplenuclide samples can be achieved where presently inhibited by interferences. This can be achieved without the need for chemical separations or efficiency tracers, thereby vastly reducing time, radioactive waste, and resulting measurement uncertainty.

Simplified kilogram traceability for high-power laser measurement using photon momentum radiometers.

Photon momentum radiometers measure the force imparted by a reflected laser beam to determine the laser's optical power. This requires high-accuracy calibration of the force sensors using milligram and microgram mass artifacts. Calibrated test masses can therefore be used to provide traceability of these radiometers to the International System of Units, but low-noise calibration at these mass levels is difficult. Here, we present the improvement in calibration capability that we have gained from implementing a robotic mass delivery system. We quantify this in terms of the specific nuances of force measurements as implemented for laser power metrology.

Direct measurement of radiation pressure and circulating power inside a passive optical cavity.

A mechanical force sensor coupled to two optical cavities is developed as a metrological tool. This system is used to generate a calibrated circulating optical power and to create a transfer standard for externally coupled optical power. The variability of the sensor as a transfer standard for optical power is less than 2%. The uncertainty in using the sensor to measure the circulating power inside the cavity is less than 3%. The force measured from the mechanical response of the sensor is compared to the force predicted from characterizing the optical spectrum of the cavity. These two forces are approximately 20% different. Potential sources for this disagreement are analyzed and discussed. The sensor is compact, portable, and can operate in ambient and vacuum environments. This device provides a pathway to novel nanonewton scale force and milliwatt scale laser power calibrations, enables direct measurement of the circulating power inside an optical cavity, and enhances the sensitivity of radiation pressure-based optical power transfer standards.

A simple method for the determination of qPlus sensor spring constants.

qPlus sensors are widely used to measure forces at the atomic scale, however, confidence in these measurements is limited by inconsistent reports of the spring constant of the sensor and complications from finite tip heights. Here we combine a numerical investigation of the force reconstruction with an experimental characterization of the flexural mechanics of the qPlus sensor. Numerical studies reveal significant errors in reconstructed force for tip heights exceeding 400 µm or one sixth of the cantilever length. Experimental results with a calibrated nanoindenter reveal excellent agreement with an Euler-Bernoulli beam model for the sensor. Prior to the attachment of a tip, measured spring constants of 1902 ± 29 N/m are found to be in agreement with theoretical predictions for the geometry and material properties of the sensor once a peaked ridge in the beam cross section is included. We further develop a correction necessary to adjust the spring constant for the size and placement of the tip.

Accurate flexural spring constant calibration of colloid probe cantilevers using scanning laser Doppler vibrometry.

Calibration of the flexural spring constant for atomic force microscope (AFM) colloid probe cantilevers provides significant challenges. The presence of a large attached spherical added mass complicates many of the more common calibration techniques such as reference cantilever, Sader, and added mass. Even the most promising option, AFM thermal calibration, can encounter difficulties during the optical lever sensitivity measurement due to strong adhesion and friction between the sphere and a surface. This may cause buckling of the end of the cantilever and hysteresis in the approach-retract curves resulting in increased uncertainty in the calibration. Most recently, a laser Doppler vibrometry thermal method has been used to accurately calibrate the normal spring constant of a wide variety of tipped and tipless commercial cantilevers. This paper describes a variant of the technique, scanning laser Doppler vibrometry, optimized for colloid probe cantilevers and capable of spring constant calibration uncertainties near ±1%.

Calculation of the effect of tip geometry on noncontact atomic force microscopy using a qPlus sensor.

In qPlus atomic force microscopy the tip length can in principle approach the length of the cantilever. We present a detailed mathematical model of the effects this has on the dynamic properties of the qPlus sensor. The resulting, experimentally confirmed motion of the tip apex is shown to have a large lateral component, raising interesting questions for both calibration and force-spectroscopy measurements.

Uncertainty quantification in nanomechanical measurements using the atomic force microscope.

Quantifying uncertainty in measured properties of nanomaterials is a prerequisite for the manufacture of reliable nanoengineered materials and products. Yet, rigorous uncertainty quantification (UQ) is rarely applied for material property measurements with the atomic force microscope (AFM), a widely used instrument that can measure properties at nanometer scale resolution of both inorganic and biological surfaces and nanomaterials. We present a framework to ascribe uncertainty to local nanomechanical properties of any nanoparticle or surface measured with the AFM by taking into account the main uncertainty sources inherent in such measurements. We demonstrate the framework by quantifying uncertainty in AFM-based measurements of the transverse elastic modulus of cellulose nanocrystals (CNCs), an abundant, plant-derived nanomaterial whose mechanical properties are comparable to Kevlar fibers. For a single, isolated CNC the transverse elastic modulus was found to have a mean of 8.1 GPa and a 95% confidence interval of 2.7-20 GPa. A key result is that multiple replicates of force-distance curves do not sample the important sources of uncertainty, which are systematic in nature. The dominant source of uncertainty is the nondimensional photodiode sensitivity calibration rather than the cantilever stiffness or Z-piezo calibrations. The results underscore the great need for, and open a path towards, quantifying and minimizing uncertainty in AFM-based material property measurements of nanoparticles, nanostructured surfaces, thin films, polymers and biomaterials.

Toggling bistable atoms via mechanical switching of bond angle.

We reversibly switch the state of a bistable atom by direct mechanical manipulation of bond angle using a dynamic force microscope. Individual buckled dimers at the Si(100) surface are flipped via the formation of a single covalent bond, actuating the smallest conceivable in-plane toggle switch (two atoms) via chemical force alone. The response of a given dimer to a flip event depends critically on both the local and nonlocal environment of the target atom-an important consideration for future atomic scale fabrication strategies.

Accurate noncontact calibration of colloidal probe sensitivities in atomic force microscopy.

The absolute force sensitivities of colloidal probes comprised of atomic force microscope, or AFM, cantilevers with microspheres attached to their distal ends are measured. The force sensitivities are calibrated through reference to accurate electrostatic forces, the realizations of which are described in detail. Furthermore, the absolute accuracy of a common AFM force calibration scheme, known as the thermal noise method, is evaluated. It is demonstrated that the thermal noise method can be applied with great success to colloidal probe calibration in air and in liquid to yield force measurements with relative standard uncertainties below 5%. Techniques to combine the electrostatics-based determination of the AFM force sensitivity with measurements of the colloidal probe's thermal noise spectrum to compute noncontact estimates of the displacement sensitivity and spring constant are also developed.

SI traceable calibration of an instrumented indentation sensor spring constant using electrostatic force.

We present a measurement scheme for creating reference electrostatic forces that are traceable to the International System of Units. This scheme yields reference forces suitable for calibrating the force sensitivity of instrumented indentation machines and atomic force microscopes. Forces between 10 and 200 muN were created and expressed in terms of the voltage, length, and capacitance between a pair of interacting electrodes. The electrodes comprised an electrically conductive sphere mounted as a tip on an instrumented indentation sensor, and a planar counterelectrode fixed to a sample stage in close proximity to the sphere. For comparison, we applied mechanical forces of similar magnitudes, first using deadweights and then using a reference force sensor. The deflection of the sensor due to the various applied forces was measured using an interferometer. A spring constant for the sensor was computed from the observed records of force versus displacement. Each procedure yielded a relative standard uncertainty of approximately 1%; however, the electrostatic technique is scalable and could provide traceable reference forces as small as a few hundred piconewtons, a range far below anything yet achieved using deadweights.

The stiffness of collagen fibrils influences vascular smooth muscle cell phenotype.

Cells receive signals from the extracellular matrix through receptor-dependent interactions, but they are also influenced by the mechanical properties of the matrix. Although bulk properties of substrates have been shown to affect cell behavior, we show here that nanoscale properties of collagen fibrils also play a significant role in determining cell phenotype. Type I collagen fibrils assembled into thin films provide excellent viewing of cells interacting with individual fibrils. Cells can be observed to extensively manipulate the fibrils, and this behavior seems to result in an incompletely spread stellate morphology and a nonproliferative phenotype that is typical of these cells in collagen gels. We show here that thin films of collagen fibrils can be dehydrated, and when seeded on these dehydrated fibrils, smooth muscle cells spread and proliferate extensively. The dehydrated collagen fibrils appear to be similar to the fully hydrated collagen fibrils in topology and in presentation of beta(1) integrin ligation sites, but they are mechanically stiffer. This decrease in compliance of dehydrated fibrils is seen by a failure of cell movement of dehydrated fibrils compared to their ability to rearrange fully hydrated fibrils and from direct measurements by nanoindentation and quantitative atomic force measurements. We suggest that increase in the nanoscale rigidity of collagen fibrils can cause these cells to assume a proliferative phenotype.

Long-term enhancement of maze learning in mice via a generalized Mozart effect.

OBJECTIVES: An animal model of the 'generalized Mozart effect' (GME) - enhanced/normalized higher brain function in response to music exposure - has been established. We extend those results in two studies using another species (mice). Study 1: (1) maze testing after music exposure was extended to a minimum of 6 hours; (2) no exposure to music in utero. Study 2: (1) music exposure time further reduced; (2) maze testing extended to 24 hours. METHODS: Study 1: two mouse groups were exposed to music continuously for 10 hours per day for 10 weeks (Group I: Mozart's Sonata K.448, Group II: Beethoven's Fur Elise). After 10 weeks, the ability to negotiate a T-maze was assessed (recording working time in maze, number of errors). Maze ability was tested 6 hours following the last music exposure. Study 2: two mouse groups were exposed periodically to music (58% silence) 10 hours per day for 10 weeks. Experiments after 10 weeks examined the groups' abilities to run the maze (recording working time/errors). Experiments were conducted 24 hours following the last music exposure. RESULTS: The Mozart group exhibited significant enhancements compared with the control mice in both studies, i.e. significantly lower working time (p<0.05) and committed fewer errors. DISCUSSION: Observation of GME in another species supports its generality for the mammalian cortex. The absence of a GME in fMRI studies for the control music also indicates a neurophysiological basis. With extended exposure, GME is a long-term effect, indicating potential clinical importance. It has been demonstrated that GME reduces neuropathological spiking significantly in epileptics. We discuss the relevance of this study for epilepsy treatment.

Innate spatial-temporal reasoning and the identification of genius.

The teaching of mathematics is invariably language-based, but spatial-temporal (ST) reasoning (making a mental image and thinking ahead in space and time) is crucial to the understanding of math. Here we report that Big Seed, a demanding ST video game, based upon the mathematics of knot theory and previously applied to understanding DNA structure and function, can be used to reveal innate ST reasoning. Big Seed studies with middle and elementary school children provide strong evidence that ST reasoning ability is not only innate but far exceeds optimistic expectations based on age, the percentage of children achieving exceptional ST performance in less than 7 h of training, and retention of ability. A third grader has been identified as a genius (functionally defined) in ST performance. Big Seed may be used for training and assessing 'creativity' (functionally defined) and ST reasoning as well as discovering genius.