The μFill: A New 96-/384- Well Microplate Reagent Dispenser for HTS and Drug Discovery

Today's biomedical research requires instrumentation that is both functional and versatile. While high throughput screening (HTS) and drug discovery laboratories require instrumentation that can be automated, pilot assay laboratories may not necessarily need total automation. Towards that end, Bio-Tek has developed the μFill, a 96-/384- well microplate reagent dispenser capable of running stand-alone or computer-controlled as part of a robotics system. The μFill is compatible with both 96- and 384-well microplates, and using a specially designed adaptor can also dispense to deep-well microplates. It is capable of dispensing from 10-3000 μl for 96- well plates and 5-1500 μl for 384-well plates in 1-μl increments. The μFill can dispense 20 μl into a 96-well plate in four seconds and into a 384-well microplate in 12 seconds. The microprocessor-controlled syringe pump is based on a tested, low-maintenance design that requires no calibration, yet provides a high degree of accuracy and precision. The accuracy with an 80 μl dispense is within 1 μl with a percentage CV of less than 2%, and with a dispense volume of 20 μl, the percentage CV is still less than 5%. For those needing to dispense organic solvents or sterile aqueous solutions, a model that is autoclavable and has increased solvent resistance is also available. The programming allows for the control of flow rates from 225 μl/well/sec, for dispensing to cell cultures, and to 1000 μl/well/sec for rapid and vigorous reagent dispensing. The flexible software provides complete programming capabilities from the keypad. For more complete automation, robotics interfaces can be developed using ActiveX® software commands. The μFill's size, with a 14 x 14-inch footprint and a height of seven inches, allows it to be used almost anywhere.     

A knowledge-based system for conveyor equipment selection

Conveyor equipment selection is a complex, and sometimes, tedious task since there are literally hundreds of equipment types and manufacturers to choose from. The expert system approach to conveyor selection provides advantages of unbiased decision making, greater availability, faster response, and reduced cost as compared to human experts. This paper discusses the development of a prototype expert system for industrial conveyor selection. The system, which was developed on Level V Object, provides the user with a list of conveyor solutions for their material handling needs along with a list of suppliers for the suggested conveyor devices. Conveyor types are selected on the basis of a suitability score, which is a measure of the fulfillment of the material handling requirements by the characteristics of the conveyor. The computation of the score is performed through the Weighted Evaluation Method, and the Expected Value Criterion for decision making under risk. The prototype system was successfully validated through two industrial case studies.     

The end of zero-hit queries: query previews for NASA’s Global Change Master Directory

The Human-Computer Interaction Laboratory (HCIL) of the University of Maryland and NASA have collaborated over three years to refine and apply user interface research concepts developed at HCIL in order to improve the usability of NASA data services. The research focused on dynamic query user interfaces, visualization, and overview + preview designs. An operational prototype, using query previews, was implemented with NASA’s Global Change Master Directory (GCMD), a directory service for earth science datasets. Users can see the histogram of the data distribution over several attributes and choose among attribute values. A result bar shows the cardinality of the result set, thereby preventing users from submitting queries that would have zero hits. Our experience confirmed the importance of metadata accuracy and completeness. The query preview interfaces make visible the problems or gaps in the metadata that are undetectable with classic form fill-in interfaces. This could be seen as a problem, but we think that it will have a long-term beneficial effect on the quality of the metadata as data providers will be compelled to produce more complete and accurate metadata. The adaptation of the research prototype to the NASA data required revised data structures and algorithms. Received: 12 December 1997 / Revised: June 1999     

Post-Placement Interconnect Entropy

We introduce the concept of post-placement interconnect entropy: the minimal number of bits required to describe a well-placed netlist, which has connection lengths distributed according to Rent's rule. The entropy is a function of the number N of cells in the netlist and the Rent exponent p. We derive an expression for the entropy per cell and show that it converges as N approaches infinity. The entropy provides an achievable lower bound on the number of configuration bits in a programmable logic device (PLD) [or field-programmable gate array (FPGA)] and a useful measure of its routing flexibility. Specific numerical values are computed for practical situations. For example, any scalable FPGA composed of 4-input lookup table cells would require 31 configuration bits per cell. We compare this to the actual number of configuration bits in a standard FPGA architecture. We generalize the bound to dimensions higher than two, and show that for any p there is an optimal dimension that minimizes the bound.     

Fatigue Threshold R‐curves Predict Fatigue Endurance Strength for Self‐Reinforced Silicon Nitride

There is a need for methods that can help predict and avoid fatigue failures of silicon nitride ceramic components. The fatigue threshold R‐curve has been proposed as potential solution to this problem. In this study, the fatigue threshold R‐curve for small, semielliptical surface cracks was calculated for a silicon nitride ceramic using the published bridging stress distribution developed from fatigue threshold tests on macroscopic crack specimens. To test the accuracy of the endurance strengths predicted using the fatigue threshold R‐curve, fatigue tests were conducted using four‐point bend beams of silicon nitride containing semielliptical surface cracks introduced by Knoop indentation. The effectiveness of the methodology was verified; indeed, 77% of the beams tested at stress levels above the predicted endurance strength failed within 10⁷ cycles and 0% of the beams tested below the predicted endurance strength failed within 10⁷ cycles. Furthermore, using the bridging stress distribution, which is thought to be a material property, the need for prohibitively difficult fatigue threshold experiments on small surface cracks is avoided. Accordingly, this methodology is potentially quite practical for use in the engineering design of ceramic mechanical components.     

Phenolic Bioactive Modulation by Lactobacillus acidophilus Mediated Fermentation of Cherry Extracts for Anti-Diabetic Functionality,Helicobacter pylori inhibition and Probiotic Bifidobacterium longum Stimulation

Cherry juice from one cultivar was fermented for 0, 24, 48, and 72 h using Lactobacillus acidophilus, and its effects related to management of hyperglycemia, hypertension, inhibition of Helicobacter pylori, and proliferation of probiotic Bifidobacterium longum were evaluated using in vitro models. Cherry extract was fermented by initially adjusting the pH to 6.0 and at the natural acidic pH. Analysis was carried out by adjusting the pH and at fermented acidic pH at each time point from each of the two samples. Total soluble phenolics decreased over a period of 72 h for initial pH adjusted samples whereas it increased or remained constant for natural acidic pH samples. DPPH linked antioxidant activity and α-glucosidase inhibitory activity decreased for samples where final pH was adjusted whereas it increased for natural acidic pH samples. Fermentation led to a decrease in hypertension-relevant ACE inhibitory activity for all samples. Natural acidic pH samples had Helicobacter pylori inhibitory activity at 24, 48 and 72 h. Based on the rationale that simple phenolics in cherry could behave as proline analogs, the potential recovery of the pathogen from inhibition was evaluated with of addition of 0.5 mM proline in the medium. A proline induced growth recovery was observed indicating that the mechanism of inhibition is related to proline dehydrogenase based oxidative phosphorylation. Overall, no inhibition was observed when samples that had H. pylori inhibitory activity were further evaluated for their effect on probiotic Bifidobacterium longum.     

Vaporization of elemental mercury from molten lead at low concentrations

Experiments were conducted to measure the rate of vaporization of elemental mercury from molten lead to provide a basis for estimating radiological source terms for the APT (Accelerator Production of Tritium project) lead blanket. These data also have application to other accelerator targets in which mercury may be created by proton spallation in lead. Molten pools of lead with from 0.01% to 0.10% mercury were prepared under inert conditions. Experiments were conducted which varied in duration from several hours to as long as a month to measure the mercury vaporization from the lead pools. The melt pools and gas atmospheres were controlled at 340 degrees C during the tests, above the melting temperature of lead. Parameters which were varied in the tests included the mercury concentrations, gas flow rates over the melt, circulation in the melts, gas atmosphere compositions and the addition of aluminum to the melts. The vaporization of mercury was found to scale roughly linearly with the concentration of mercury in the pool. Variations in the gas flow rates were not found to have any effect on the mass transfer, however circulation of the melt by a submerged stirrer did enhance the mercury vaporization rate. The rate of mercury vaporization under a high-purity argon atmosphere was found to exceed that for an air atmosphere by as much as a factor of from ten to 20; the causal factor in this variation was the formation of an oxide layer over the melt pool with the air atmosphere which retarded mass transfer across the melt-atmosphere interface. Aluminum was introduced into the melt to investigate its effect upon the mercury vaporization rate. No effect was observed for a case under a high-purity argon atmosphere, which suggests that there are no chemical effects of the aluminum on the vaporization kinetics. With an air atmosphere, the presence of aluminum in the melt reduced the mercury vaporization by a factor of six in comparison to the identical test but without aluminum, suggesting that aluminum in the lead/ mercury melt retards the vaporization of mercury by creating a surface oxide layer in addition to the lead-oxide layer or by changing the character of the lead-oxide layer, thereby increasing the mass transfer resistance.     

Bendable bulk metallic glass: Effects of a thin,adhesive, strong,and ductile coating

We demonstrate, for the first time, that a thin, strong, ductile, and adhesive coating renders bulk metallic glasses (BMGs) bendable. The bending ductility of 3 mm thick BMGs, Zr₅₀Cu₃₀Al₁₀Ni₁₀ in this case, can be dramatically enhanced from ～0% to ～13.7% by the deposition of a thin bilayer film on the tensile side of the BMG sample. The bilayer, consisting of a 25 nm thick Ti adhesive layer with a 200 nm thick metallic glass (MG) overlayer, exhibits the required synergistic combination of good adhesion, high strength, and ductility compared with other single-layer films examined (Ti, TiN, and MG). Cross-sectional scanning and transmission electron microscopy, together with finite element modeling, reveal that the bilayer coating absorbs deformation while allowing more homogeneous formation of a high density of smaller shear bands at the bilayer/BMG interface. The bilayer coating, in turn, covers surface weak points and minimizes the formation of localized shear bands which lead to catastrophic failure under bending. As a result, the average shear-band spacing in bilayer-coated BMGs is small, 54 μm, and approximately equal to that found in bendable, 450 μm thick, MG ribbons. Thus, coated BMGs can accommodate large strains and overcome the MG size effect, without sacrificing their extraordinary mechanical properties. Our results for both coated and uncoated BMGs, as well as previously reported results for uncoated metallic glasses, with thicknesses ranging from ribbons to thin plates to bulk, are well described by a simple power law relationship between plastic strain to failure and shear band spacing. This scaling law may be useful in guiding future experiments toward producing more flexible BMGs.