Resolving neurotransmitters detected by fast-scan cyclic voltammetry

Carbon-fiber microelectrodes are frequently used as chemical sensors in biological preparations. In this work, we evaluated the ability of cyclic voltammograms recorded at fast-scan rates to resolve neurochemicals when analyzed by principal component regression. A calibration set of 30 cyclic voltammograms was constructed from 9 different substances at a variety of concentrations. The set was reduced by principal component analysis, and it was found that 99.5% of the variance in the data could be captured with five principal components. This set was used to evaluate cyclic voltammograms obtained with one or two compounds present in solution. In most cases, satisfactory predictions of the identity and concentration of analytes were obtained. Chemical dynamics were also resolved from a set of fast-scan cyclic voltammograms obtained with the electrode implanted in a region of a brain slice that contains dopaminergic terminals. Following stimulation, principal component regression of the data resolved the changes in dopamine and pH that were evoked. In a second test of the method, vesicular release was measured from adrenal medullary cells and the data were evaluated with a calibration set composed of epinephrine and norepinephrine. Cells that secreted one or the other were identified. Overall, the results show that principal component regression with appropriate calibration data allows resolution of substances that give overlapping cyclic voltammograms.     

Spatially and temporally resolved single-cell exocytosis utilizing individually addressable carbon microelectrode arrays

We report the fabrication and characterization of carbon microelectrode arrays (MEAs) and their application to spatially and temporally resolve neurotransmitter release from single pheochromocytoma (PC12) cells. The carbon MEAs are composed of individually addressable 2.5-mum-radius microdisks embedded in glass. The fabrication involves pulling a multibarrel glass capillary containing a single carbon fiber in each barrel into a sharp tip, followed by beveling the electrode tip to form an array (10-20 microm) of carbon microdisks. This simple fabrication procedure eliminates the need for complicated wiring of the independent electrodes, thus allowing preparation of high-density individually addressable microelectrodes. The carbon MEAs have been characterized using scanning electron microscopy, steady-state and fast-scan voltammetry, and numerical simulations. Amperometric results show that subcellular heterogeneity in single-cell exocytosis can be electrochemically detected with MEAs. These ultrasmall electrochemical probes are suitable for detecting fast chemical events in tight spaces, as well as for developing multifunctional electrochemical microsensors.     

Computing Low Latency Batches with Unreliable Workers in Volunteer Computing Environments

Internet based volunteer computing projects such as SETI@home are currently restricted to performing coarse grained, embarrassingly parallel master-worker style tasks. This is partly due to the “pull” nature of task distribution in volunteer computing environments, where workers request tasks from the master rather than the master assigning tasks to arbitrary workers. In this paper we propose algorithms for computing batches of medium grained tasks with deadlines in pull-style volunteer computing environments. We develop models of unreliable workers based on analysis of trace data from an actual volunteer computing project. These models are used to develop algorithms for task distribution in volunteer computing systems with a high probability of meeting batch deadlines. We develop algorithms for perfectly reliable workers, computation-reliable workers and unreliable workers. Finally, we demonstrate the effectiveness of the algorithms through simulations using traces from actual volunteer computing environments.     

Voltammetric detection of metal nanoparticles separated by liquid chromatography

Fast-scan cyclic voltammetric (FSCV) detection of the separation of small (< 2-nm core diameter) organothiolate monolayer-protected gold clusters (MPCs) by high-performance liquid chromatography (HPLC) is demonstrated in this paper. Cyclic voltammograms taken at 1-s intervals (scan rate 150 V/s) display distinctive current-potential patterns for different MPCs, aiding in their identification. Previous amperometric detection of MPC separations required multiple HPLC runs at different electrode potentials to construct hydrodynamic current-potential voltammograms. Theory for ratiometric response of photodiode array absorbance and constant potential amperometry detectors was further tested on the fast-scan voltammetric detector and gave reasonable relative core size results for a mixture of Au140 and Au38 MPCs. In the first analysis of mixed MPC monolayer composition that is not simply an overall average result, preliminary HPLC experiments on MPCs with mixed hexanethiolate and ferrocenyl-octanethiolate monolayers resolved peaks that are shown to correspond to different average numbers of ferrocene-labeled ligands per MPC.     

Probing electric fields inside microfluidic channels during electroosmotic flow with fast-scan cyclic voltammetry

Fast-scan cyclic voltammetry (FSCV) at carbon-fiber microelectrodes was used in microfluidic channels. This method offers the advantage that it can resolve electroactive species not separated in the channel. In addition, this method provides a route to investigate the distribution of applied electrophoretic fields in microfluidic channels. To probe this, microelectrodes were inserted at various distances into channels and cyclic voltammograms recorded at 300 V/s were repeated at 0.1-s intervals. The use of a battery-powered laptop computer and potentiostat provided galvanic isolation between the applied electrophoretic field and the electrochemical measurements. In the absence of an external field, the peak potential for oxidation of the test solute, Ru(bpy)3(2+), was virtually unaltered by insertion of the microelectrode tip into the channel. When an electrophoretic field was applied, the peak potential for Ru(bpy)3(2+) oxidation shifted to more positive potentials in a manner that was directly proportional to the field in the channel. The shifts in peak potential observed with FSCV enabled direct compensation of the applied electrochemical potential. This approach was used to explore the electrophoretic field at the channel terminus. It was found to persist for more than 50 microm from the channel terminus. In addition, the degree of analyte dispersion was found to depend critically on the electrode position outside the channel.     

MS/MS methodology to improve subcellular mapping of cholesterol using TOF-SIMS

Time-of-flight secondary ion mass spectrometry (TOF-SIMS) can be utilized to map the distribution of various molecules on a surface with submicrometer resolution. Much of its biological application has been in the study of membrane lipids, such as phospholipids and cholesterol. Cholesterol is a particularly interesting molecule due to its involvement in numerous biological processes. For many studies, the effectiveness of chemical mapping is limited by low signal intensity from various biomolecules. Because of the high energy nature of the SIMS ionization process, many molecules are identified by detection of characteristic fragments. Commonly, fragments of a molecule are identified using standard samples, and those fragments are used to map the location of the molecule. In this work, MS/MS data obtained from a prototype C60(+)/quadrupole time-of-flight mass spectrometer was used in conjunction with indium LMIG imaging to map previously unrecognized cholesterol fragments in single cells. A model system of J774 macrophages doped with cholesterol was used to show that these fragments are derived from cholesterol in cell imaging experiments. Examination of relative quantification experiments reveals that m/z 147 is the most specific diagnostic fragment and offers a 3-fold signal enhancement. These findings greatly increase the prospects for cholesterol mapping experiments in biological samples, particularly with single cell experiments. In addition, these findings demonstrate the wealth of information that is hidden in the traditional TOF-SIMS spectrum.     

Temporal resolution in electrochemical imaging on single PC12 cells using amperometry and voltammetry at microelectrode arrays

Carbon-fiber-microelectrode arrays (MEAs) have been utilized to electrochemically image neurochemical secretion from individual pheochromocytoma (PC12) cells. Dopamine release events were electrochemically monitored from seven different locations on single PC12 cells using alternately constant-potential amperometry and fast-scan cyclic voltammetry (FSCV). Cyclic voltammetry, when compared to amperometry, can provide excellent chemical resolution; however, spatial and temporal resolution are both compromised. The spatial and temporal resolution of these two methods have been quantitatively compared and the differences explained using models of molecular diffusion at the nanogap between the electrode and the cell. A numerical simulation of the molecular flux reveals that the diffusion of dopamine molecules and electrochemical reactions both play important roles in the temporal resolution of electrochemical imaging. The simulation also reveals that the diffusion and electrode potential cause the differences in signal crosstalk between electrodes when comparing amperometry and FSCV.     

Accelerating earthquake simulations on general‐purpose graphics processors

Parallelization strategies are presented for Virtual Quake, a numerical simulation code for earthquakes based on topologically realistic systems of interacting earthquake faults. One of the demands placed upon the simulation is the accurate reproduction of the observed earthquake statistics over three to four decades. This requires the use of a high‐resolution fault model in computations, which demands computational power that is well beyond the scope of off‐the‐shelf multi‐core CPU computers. However, the recent advances in general‐purpose graphic processing units have the potential to address this problem at moderate cost increments. A functional decomposition of Virtual Quake is performed, and opportunities for parallelization are discussed in this work. Computationally intensive modules are identified, and these are implemented on graphics processing units, significantly speeding up earthquake simulations. In the current best case scenario, a computer with six graphics processing units can simulate 500 years of fault activity in California at 1.5 km × 1.5 km element resolution in less than 1 hour, whereas a single CPU requires more than 2 days to perform the same simulation. Copyright © 2015 John Wiley & Sons, Ltd.