Sulfide-rich materials comprising the waste at the abandoned Montalbion silver mine have undergone extensive oxidation prior to and after mining. Weathering has led to the development of an abundant and varied secondary mineral assemblage throughout the waste material. Post-mining minerals are dominantly metal and/or alkali (hydrous) sulfates, and generally occur as earthy encrustations or floury dustings on the surface of other mineral grains. The variable solubility of these efflorescences combined with the irregular rainfall controls the chemistry of seepage waters emanating from the waste dumps. Irregular rainfall events dissolve the soluble efflorescences that have built up during dry periods, resulting in 'first-flush' acid (pH 2.6-3.8) waters with elevated sulfate, Fe, Cu and Zn contents. Less-soluble efflorescences, such as anglesite and plumbojarosite, retain Pb in the waste dump. Metalrich (Al, Cd, Co, Cu, Fe, Mn, Ni, Zn) acid mine drainage waters enter the local creek system. Oxygenation and hydrolysis of Fe lead to the formation of Fe-rich precipitates (schwertmannite, goethite, amorphous Fe compounds) that, through adsorption and coprecipitation, preferentially incorporate As, Sb and In. Furthermore, during dry periods, evaporative precipitation of hydrous alkali and metal sulfate efflorescences occurs on the perimeter of stagnant pools. Flushing of the streambed by neutral pH waters during heavy rainfall events dissolves the efflorescences resulting in remobilisation and transport of sulfate and metals (particularly Cd, Zn) downstream. Thus, in areas of seasonal or irregular rainfall, secondary efflorescent minerals present in waste materials or drainage channels have an important influence on the chemistry of surface waters.
A coustic energy can precisely and accurately eject a droplet of liquid from a reservoir, enabling delivery of picoliter and nanoliter volumes (Ellson, R.; Mutz, M.; Browning, B.; Lee, L.; Miller, M.; Papen, R. Transfer of low nanoliter volumes between microplates using focused acousticsdautomation considerations. Journal of the Association for Laboratory Automation 2003, 8(5), 29e34). Acoustic droplet ejection has been shown to be extremely precise (coefficients of variation !2%) over a wide range of dispensed volumes (Ellson, R.; Mutz, M.; Browning, B.; Lee, L.; Miller, M.; Papen, R. Transfer of low nanoliter volumes between microplates using focused acousticsdautomation considerations. Journal of the Association for Laboratory Automation 2003, 8(5), 29e34). However, measuring the performance of low-volume fluid transfers can be difficult because the data are often masked by variability in bulk dispensers and fluorescence readers used as part of the overall measurement process (Petersen, J.; Nguyen, J. Comparison of absorbance and fluorescence methods for determining liquid dispensing precision. Journal of the Association for Laboratory Automation 2005, 10(2), 82e87; Rhode, H.; Schulze, M.; Renard, S.; Zimmerman, P.; Moore, T.; Cumme, G.; Horn, A. An improved method for checking HTS/uHTS liquid handling systems. Journal of Biomolecular Screening 2004, 9, 726e733). The fluorophore used must also be stable so that thermal bleaching and photobleaching do not contribute additional variability to the measurements. This study assesses the suitability of fluorescein to measure the precision of fluid transfers of 2.5-nL DMSO droplets. The short-term and long-term stabilities of fluorescein are first qualified using a reference standard. Next, we determine the noise contribution of the filler and reader. Lastly, data are presented for the precision of 5-and 50-nL fluid transfers using this fluorescein measurement process. ( JALA 2006;11:233-9)
More accurate dose-response curves can be constructed by eliminating aqueous serial dilution of compounds. Traditional serial dilutions that use aqueous diluents can result in errors in dose-response values of up to 4 orders of magnitude for a significant percentage of a compound library. When DMSO is used as the diluent, the errors are reduced but not eliminated. The authors use acoustic drop ejection (ADE) to transfer different volumes of model library compounds, directly creating a concentration gradient series in the receiver assay plate. Sample losses and contamination associated with compound handling are therefore avoided or minimized, particularly in the case of less water-soluble compounds. ADE is particularly well suited for assay miniaturization, but gradient volume dispensing is not limited to miniaturized applications.
Acoustic droplet ejection (ADE) enables crystallization experiments at the low-nanoliter scale, resulting in rapid vapor diffusion equilibration dynamics and efficient reagent usage in the empirical discovery of structure-enabling protein crystallization conditions. We extend our validation of this technology applied to the diverse physicochemical property space of aqueous crystallization reagents where dynamic fluid analysis coupled to ADE aids in accurate and precise dispensations. Addition of crystallization seed stocks, chemical additives, or small-molecule ligands effectively modulates crystallization, and we here provide examples in optimization of crystal morphology and diffraction quality by the acoustic delivery of ultra-small volumes of these cofactors. Additional applications are discussed, including set up of in situ proteolysis and alternate geometries of crystallization that leverage the small scale afforded by acoustic delivery. Finally, we describe parameters of a system of automation in which the acoustic liquid handler is integrated with a robotic arm, plate centrifuge, peeler, sealer, and stacks, which allows unattended high-throughput crystallization experimentation.
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