Development of an In-Source Peltier Heater for Pulled Capillary Nanospray Emitter Columns.

Column compartments in liquid chromatography (LC) systems house the LC columns. These compartments are responsible for maintaining a suitable column environment for achieving optimal chromatographic performance. However, the advancements in instrument and column designs demand newer technologies. It is a well-established concept that decreasing the dead volume of the column improves the column resolution, thereby providing enhanced chromatographic separation. One of the major contributors in the dead volume is the line connecting the column in the LC compartment to the ion source in the mass spectrometer. Using in-source emitter columns is one strategy to enhance the resolution. However, ion sources without temperature control are not suitable for columns that are used at high temperatures. In this work, we are introducing a nano electrospray ionization source with an integrated Peltier heater designed for pulled capillary nanospray emitter columns. Although the performance of the device is demonstrated by showing the isomeric separation of permethylated glycans using a mesoporous graphitized carbon packed pulled capillary emitter, it can easily be paired with any nanospray emitter column that requires temperature control.

Probing hypoxia-induced staurosporine resistance in prostate cancer cells with a microfluidic culture system.

A microfluidic system for cell culture and drug response studies was developed to elucidate the effects of hypoxia on drug susceptibility. Drug response studies were performed in prostate cancer cells and Ramos B cells under normoxic and hypoxic conditions. A vacuum actuated microfluidic culture device was used for cell culture and PC3 cells were cultured in the chip up to 16 hours. Cells were treated with several concentrations of staurosporine and apoptosis was assayed using the fluorescent probes MitoTracker Deep Red and Annexin-V. For hypoxic samples, the chip was placed in a hypoxia chamber and pre-conditioned at <1% oxygen before inducing the cells with staurosporine. Cells exposed to 2 µM staurosporine were 32% ± 10% apoptotic under normoxic conditions but only 1.5% ± 12% apoptotic under hypoxic conditions. As little as 1 hour of hypoxic preconditioning increased drug resistance. Cell apoptosis correlated with drug dose, although in each case hypoxia reduced the apoptotic fraction significantly. Given the rapid nature of cell adaptation to hypoxia, this chip and analysis approach can be used to identify compounds that can induce cell death in hypoxic tumor cells rapidly.

Characterization of a novel particle into liquid sampler for analysis of single fluorescent aerosol particles through capillary electrophoresis.

An approach to sample and analyze single aerosolized droplets (<10 nL) of solutions containing fluorescein isothiocyanate (FITC) labeled glycine (GLY) and glutamic acid (GLU) is demonstrated. The sampling approach is based on inertial impaction in which the sample particle is accelerated through a nozzle and directly into a small drop of buffered solution (20 mM borate, pH=10) suspended at the end of a coaxial tube of stainless steel and a fused silica capillary. A spherical light scattering cell and laser (λ=532 nm) is used to detect the arrival of particles at the buffered droplet. Upon dissolution and/or mixing, a portion of the sample is injected onto the fused silica capillary for subsequent chemical analysis by capillary electrophoresis (CE) and detection by laser-induced fluorescence (LIF). It was found that the inertial impaction approach sampled particles >1 µm diameter with an efficiency of 80% or greater. At 15 kV applied potential, the FITC conjugates of GLY and GLU could be resolved in less than 120 s allowing qualitative analysis of the contents of single dispersed particles. However, the extent to which the sample is diluted into the buffer droplet varied significantly on a per-particle basis that caused >80% R.S.D. in fluorescence peak heights. This aspect of the method would necessitate the use of internal standards for quantitative analysis of materials present within the particles. It is envisaged that further improvements to the device described may ultimately lead to analysis of the contents of single particles dispersed in earth's atmosphere.

Simultaneous measurement of optical scattering and extinction on dispersed aerosol samples.

Accurate and precise measurements of light scattering and extinction by atmospheric particulate matter aid understanding of tropospheric photochemistry and are required for estimates of the direct climate effects of aerosols. In this work, we report on a second generation instrument to simultaneously measure light scattering (b(scat)) and extinction (b(ext)) coefficient by dispersed aerosols. The ratio of scattering to extinction is known as the single scatter albedo (SSA); thus, the instrument is referred to as the albedometer. Extinction is measured with the well-established cavity ring-down (CRD) technique, and the scattering coefficient is determined through collection of light scattered from the CRD beam. The improved instrument allows reduction in sample volume to <1% of the original design, and a reduction in response time by a factor of >30. Through using a commercially available condensation particle counter (CPC), we have measured scattering (σ(scat)) and extinction (σ(ext)) cross sections for size-selected ammonium sulfate and nigrosin aerosols. In most cases, the measured scattering and extinction cross section were within 1 standard deviation of the accepted values generated from Mie theory suggesting accurate measurements are made. While measurement standard deviations for b(ext) and b(scat) were generally <1 Mm(-1) when the measurement cell was sealed or purged with filtered air, relative standard deviations >0.1 for these variables were observed when the particle number density was low. It is inferred that statistical fluctuations of the absolute number of particles within the probe beam leads to this effect. However, measured relative precision in albedo is always superior to that which would be mathematically propagated assuming independent measurements of b(scat) and b(ext). Thus, this report characterizes the measurement precision achieved, evaluates the potential for systematic error to be introduced through light absorption by gases, presents comparisons with Mie theory, and provides ambient monitoring data collected on a mineral dust dominated aerosol at our location.