Light sheet based imaging flow cytometry on a microfluidic platform

We propose a light sheet based imaging flow cytometry technique for simultaneous counting and imaging of cells on a microfluidic platform. Light sheet covers the entire microfluidic channel and thus omits the necessity of flow focusing and point scanning based technology. Another advantage lies in the orthogonal detection geometry that totally cuts‐off the incident light, thereby substantially reducing the background in the detection. Compared to the existing state‐of‐art techniques the proposed technique shows marked improvement. Using fluorescently‐coated Saccharomyces cerevisiae cells we have recorded cell counting with throughput as high as 2,090 cells/min in the low flow rate regime and were able to image the individual cells on‐the‐go. Overall, the proposed system is cost‐effective and simple in channel geometry with the advantage of efficient counting in operational regime of low laminar flow. This technique may advance the emerging field of microfluidic based cytometry for applications in nanomedicine and point of care diagnostics. Microsc. Res. Tech. 76:1101–1107, 2013. © 2013 Wiley Periodicals, Inc.     

Numerical analysis of the subsonic flow around a three-dimensional cavity

Flight vehicles such as wheel wells and bomb bays have many cavities. The flow around a cavity is characterized as an unsteady flow because of the formation and dissipation of vortices brought about by the interaction between the free stream shear layer and the internal flow of the cavity. The resonance phenomena can damage the structures around the cavity and negatively affect the aerodynamic performance and stability of the vehicle. In this study, a numerical analysis was performed for the cavity flows using the unsteady compressible three-dimensional Reynolds-Averaged Navier-Stokes (RANS) equation with Wilcox’s turbulence model. The Message Passing Interface (MPI) parallelized code was used for the calculations by PC-cluster. The cavity has aspect ratios (L/D) of 2.5 ∼ 7.5 with width ratios (W/D) of 2 ∼ 4. The Mach and Reynolds numbers are 0.4 ∼ 0.6 and 1.6×10 ⁶ , respectively. The occurrence of oscillation is observed in the “shear layer and transient mode” with a feedback mechanism. Based on the Sound Pressure Level (SPL) analysis of the pressure variation at the cavity trailing edge, the dominant frequencies are analyzed and compared with the results of Rossiter’s formula. The dominant frequencies are very similar to the result of Rossiter’s formula and other experimental data in the low aspect ratio cavity (L/D = ∼ 4.5). In the large aspect ratio cavity, however, there are other low dominant frequencies due to the leading edge shear layer with the dominant frequencies of the feedback mechanism. The characteristics of the acoustic wave propagation are analyzed using the Correlation of Pressure Distribution (CPD). This paper was recommended for publication in revised form by Associate Editor Do Hyung Lee Hong-il CHOI received the B.S and M.S degrees in Aerospace Engineering from Chosun University, Korea in 2005 and 2008, respectively. He currently work at KOREA Electric Power Research Institute in Korea Pa-ul MUN received the B.S in Aerospace Engineering from Chosun University, Korea in 2008. He is currently Candidate for the degree of master of Aerospace Engineering at Chosun University in Korea. Jae-soo KIM received the B.S in Aerospace Engineering from Seoul National University in Korea in 1981. He then received his M.S and Ph.D. degree in Aerospace Engineering from KAIST in Korea in 1983 and 1987, respectively. He spent one year at Cornell university(USA) as a Post Doc. He worked at Korea Aerospace Research Institute for eight years. Dr. Kim is currently a Professor at the Department of Aerospace Engineering at Chosun University in Korea     

The value of assessing cell proliferation in breast cancer

Using three methods to measure cell proliferation, namely DNA cell cycle; anti-proliferating cell monoclonal antibody (MAb) (Ki67, P145) analysis by flow cytometry; and the histological silver (argyrophilic) staining technique to visualize nuclear-organizing regions (AgNOR), twenty-two paired samples of primary breast tumour and axillary lymph node were analysed. The results showed variable levels of correlation between the methods used for the tumour group (r = 0.915, P <0.001 for Ki677 versus P145; r = 0.42, P <0.005 for percentage S/G2/M-phase versus P145; r = 0.16, P <0.5 for percentage S/G2/M-phase versus AgNOR; r = 0.400, P < 0.1 for Ki67 versus AgNOR). The lymph-node group showed slightly poorer correlations, yet involved nodes showed a consistently higher level of proliferation than non-involved nodes by all methods used. Overall, MAb binding of Ki67 or P145 was seen to be a good indicator of cycling cells, detecting G1-phase cells in addition to S/G2/M-phase cells indentified by the other methods used. However, no advantage was found over the usual DNA flow cytometric analysis of cells, which had clear prognostic value. AgNOR scores were found to be able to discriminate between diploid and aneuploid; and dividing and non-dividing cells, but areas of score overlap limited the application of this technique to that of a positive discriminator only.     

DETERMINATION OF FLAVONES IN LOPHATHERUM GRACILE BY LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY

Lophatherum gracile Brongn. (L. gracile) has been used for food and medicine in China for thousands of years. Flavonoids are one of the main bioactive components of this herb. A reliable method was established using liquid chromatography with tandem mass spectrometry to determine the concentrations of seven flavonoids. The separation was performed using a C₁₈ column at 40°C and the mobile phase was a mixture of methanol and 0.3% formic acid (v/v) in water in gradient elution mode at a flow rate of 1 mL/min. Quantification was performed through tandem mass spectrometry with negative electrospray ionization and multiple reaction monitoring at m/z [M-H]^?. The method was validated using calibration curves, limits of detection and quantification, precision, repeatability, stability, and accuracy. Seven compounds in eleven samples were determined, and there were significant differences in the concentrations of isoorientin, swertiajaponin, afzelin, luteolin-7-O-β-D-glucoside, vitexin, isovitexin, and swertisin. The established method is suitable for quality control for the analysis of L. gracile, as well as to provide necessary information for the rational utilization of L. gracile resources.     

The development of protein chip using protein G for the simultaneous detection of various pathogens

A protein chip using protein G for the simultaneous detection of various pathogens such as Escherichia coli O157:H7, Salmonella typhimurium, Yersinia enterocolitica, and Legionella pneumophila was developed. In order to endow the orientation of antibody molecules on solid surface, protein G was introduced. The protein G on gold (Au) surface modified with 11-mercaptoundecanoic acid (MUA) was arrayed and then four different kinds of monoclonal antibodies (Mabs) against pathogens (E. coli O157:H7, S. typhimurium, Y. enterocolitica, and L. pneumophila) on protein G spots were selectively arrayed using a microarrayer, and its spatial density was over 2400 spots cm². Using the constructed protein chip, the various pathogens such as E. coli O157:H7, S. typhimurium, Y. enterocolitica, and L. pneumophila could be detected by a sandwich method and its lowest detection limit for E. coli O157:H7 was 10² CFU/ml. The proposed fabrication technique of protein chip for the detection of various pathogens could be applied to construct other protein chips with a high efficiency.     

Insight of EDX analysis and EFTEM: Are spherocrystals located in strombidae digestive gland implied in detoxification of trace metals?

Digestive tubules of Strombidae are composed by three cell types: digestive cells, vacuolated cells, and crypt cells. The last one is characterized by the presence of intracellular granules identified as spherocrystals. Such structures are known to occur in basophilic cells of gastropod digestive gland, where they are supposed to be involved in the regulation of some minerals and in detoxification. In this study, energy‐dispersive X‐ray analysis (EDX) and energy filtered transmission electron microscopy (EFTEM) were used to determine the elemental content of spherocrystals in two Strombidae, Strombus gigas and Strombus pugilis. In freshly collected individuals of both species, the following elements were detected: Ca, Fe, Mg, P, and Zn. Aluminum and Mn were also detected in S. gigas. Their presence in spherocrystals indicates that, in Strombidae, spherocrystals are involved in the regulation of minerals and essential trace metals. In order to answer the question “are spherocrystals involved in nonessential trace metals scavenging?,” artificial cadmium and lead exposure by both waterborne and dietary pathways was applied to S. pugilis. No evidence of cadmium (Cd(NO₃)₂) or lead (Pb(NO₃)₂) provided by food was found in spherocrystals. Cadmium provided in water (Cd(NO₃)₂ and CdCl₂) causes structural modifications of the digestive gland; however, this element was not trapped in spherocrystals. These results suggest that spherocrystals are not involved in detoxification of such nonessential trace metals. Microsc. Res. Tech., 2011. © 2011 Wiley‐Liss, Inc.     

Experimental and numerical study for the cross-flow around four cylinders in an in-line square configuration

The cross-flow around four cylinders in an in-line square configuration with the spacing ratio (L/D) 1.5, 2.5, 3.5 and 5.0 have been investigated experimentally using Laser Doppler Anemometer (LDA) and Digital Particle Image Velocimetry (DPIV). The experiments were carried out in a closed-loop wind tunnel with Reynolds number 1.128× 10⁴ to 1.982× 10⁴. Mean velocity distributions are obtained by LDA. The full field instantaneous and averaged velocity and vorticity components are measured by DPIV The present experimental study indicated that several distinct flow patterns exist. Distinct vortex shedding of the upstream cylinders was suppressed forL/D < 3.5 atRe=1.128× 10⁴. The flow patterns are affected by the spacing ratio andRe. In order to capture the details of the 3-D vortices structures and obtain all the instantaneous physical information, 3-D numerical simulations of the cross-flow around the four cylinders in an in-line square configuration with the spacing ratio 1.5 and 3.5, andRe=1.50× 10⁴ are carried out using large eddy simulation (LES). The numerical results are in good agreement with the experimental results. These results provided full field instantaneous information of the flow structures, velocity field and vorticity field of cross-flow around the four cylinders in an in-line square configuration.     

Mapping cholesteryl ester analogue uptake and intracellular flow in Paramecium by confocal fluorescence microscopy

In Paramecium primaurelia the uptake and intracellular flow of cholesteryl ester was studied by fluorescence confocal laser scanning optical microscopy and by the fluorescent analogue cholesteryl‐BODIPY^® FL C₁₂ (BODIPY‐CE). The BODIPY FL fluorophore has the characteristic of emitting green fluorescence, which is red‐shifted as the probe concentrates. In cells incubated with 25 µm BODIPY‐CE for 30 s, fluorescence is found in vesicles located around the cytopharynx in the posterior half of the cell. Successively, the lipid is internalized by food vacuoles, the fluorescent vesicles are distributed throughout the cell and the intracellular membranes are labelled. The food vacuole number is maximum after 10–15 min of continuous labelling, then it decreases until no food vacuoles are found in 30‐min fed cells. BODIPY‐CE accumulates in red‐labelled cytoplasmic droplets located in the anterior half of the cell. When food vacuole formation is inhibited by trifluoperazine, fluorescence is found on cellular membranes and in small green‐labelled vesicles at the apical pole. The inhibition of clathrin‐mediated endocytosis does not interfere in P. primaurelia with BODIPY‐CE intracellular flow: intracellular membranes and storage droplets in the cell anterior part are dyed. Conversely, the use of sterol‐binding drugs prevents the lipid accumulation in droplets, stopping the lipid within the cytoplasmic membranes. Furthermore, the cells treated with monensin and cytochalasin B show a labelling of the cellular membranes and lipid droplets, whereas NH₄Cl reduces the lipid storage. Low temperature (4 °C) does not prevent the internalization of BODIPY‐CE that, however, is localized at the cytoplasmic membrane level and does not accumulate in storage droplets. In addition, BODIPY‐CE inhibits phagocytosis, as evidenced by comparing the kinetics of food vacuole formation of control cells, only fed with latex particles, with that of cells fed with latex particles and BODIPY‐CE. In conclusion, this study points out that in P. primaurelia the cholesteryl ester enters the cell via food vacuoles and through the plasma membrane and, inside the cell, it alters cell functions.     

Comparison of some physical techniques for detection of spoilage in apple juice inoculated with Saccharomyces cerevisiae: Optical and photothermal methods

Abstract

Several physical techniques were used to study the extent of spoilage in apple juice deliberately inoculated with yeast (concentration of Saccharomyces cerevisiae ranged from 25 cells mL^?1 to 2.5 × 10⁶ cells mL^?1, respectively) and their performance compared in terms of detection limit achieved. The optical methods used in this investigation rely on the measurement of either absorption [as is the case for classical spectrophotometry (SP) and the so called optothermal window (OW), a variant of a photothermal method], or scattering [examples are turbidimetry (TB), laser scattering (SC), and laser speckle fluctuation (SF)]. It is shown that the presence of yeast increases both optical absorption and scattering. The most favorable detection limit (25 cells mL^?1) and a highest (nearly 10⁴) dynamic range, combined with a good linearity, were obtained with the experimental set‐up for SC. In addition, the extent of correlation between different methods was determined using two markedly different reference substances, i.e., (i) the mixture of apple and blackcurrant juices, representing a strongly absorbing sample, and (ii) diluted (dilution factor of 10³) milk as a strong scatterer. Finally, one has monitored the progress of a spontaneous spoilage process in the inoculated juices stored at 5°C under aerobic conditions.     

Experimental studies on heat transfer in the annuli with corrugated inner tubes

Experimental heat transfer data for single-phase water flow in the annuli with corrugated inner tubes are presented. In the annuli with parallel flow, ten different annular arrangements are considered. For water (low rate in l,700<Re<13,000 regime, data for Nusselt numbers are presented. The results show significant effects of both the pitch to trough height ratio (P/e) and the radius ratio (r ^*). AsP/e becomes closer to 8 in the range below the radius ratio(r ^*) of 0.5. Nusselt numbers increase. However, Nusselt numbers decrease in the range above the radius ratio (r^*) of 0.5 because flow reattachment position becomes farther in the narrower clearance.     

Numerical investigation on Rayleigh-Bénard-Marangoni flow of water near the maximum density in cylindrical pools with large aspect ratios

We report a set of direct numerical simulation results on Rayleigh-Bénard-Marangoni (R-B-M) flow of cold water in the cylindrical pools. The heat transfer between the free surface and the environment is considered. The aspect ratio Г of the cylindrical pool varies from 2 to 8. Rayleigh (Ra) and Biot (Bi) numbers are respectively confined in Ra ≤ 10⁴ and 0 < Bi ≤ 50. The flow onset critical Ra is determined and the influences of Bi and the density inversion parameter (Θ_m) on the critical Ra are analyzed. The primary bifurcation flow structures of R-B-M flow are shown and the evolution of the flow structures with Ra and Bi at different Θ_m is observed. Furthermore, the heat transfer ability is estimated by Nusselt number. The results indicate that the critical Ra of the flow onset increases with increasing Bi and Θ_m. But it decreases with the increase of Г. The primary bifurcation pattern is multicellular flow. With increasing Г, the number of flow cells in multicellular flow increases fast. With increasing Ra, the up-triangular and up-quadrilateral flow structures appear at Г = 4, and finally transits to the up one-torus. With increasing Ra and Г, and decreasing Θ_m, average Nusselt number increases monotonically. However, with increasing Bi, it first increases, and then decreases.

     

An improved method for predicting discharge of homogeneous compound channels based on energy concept

Accurate estimation of flow discharge in a compound river channel is increasingly important in river management and hydro-environment design. In this paper, a new model is developed to improve the prediction of flow based on Energy Concept Method (ECM) and Weighted Divided Channel Method (WDCM) along with the apparent shear stress at the interface between main channel and floodplain. The new model is compared with a wide range of our experimental data and the data available in the literature. The 27 datasets used include homogenous symmetric channels (22 datasets) and asymmetric channels (5 datasets) with various aspect ratios [channel total width (B) at bankfull / main channel bottom (b) =1.5–15.8], and bed slopes (S_o = 4.3 × 10⁻⁴–1.3 × 10⁻²). It was found that the new model has significantly improved the accuracy of flow prediction compared with the traditional Divided Channel Method (DCM), and has also considerably better results than the ECM and WDCM methods against all the datasets, particularly for relatively low flow depths of floodplain where the flow discharges are most difficult to predict correctly. The new model predicts the total discharge well for both symmetric and asymmetric channels, within an averaged relative error of about 5%.     

Characterization of biological macromolecules by combined mass mapping and electron energy-loss spectroscopy

The combination of scanning transmission electron microscopy (STEM) and parallel-detection energy-loss spectroscopy (EELS) was used to detect specific bound elements within macromolecules and macromolecular assemblies prepared by direct freezing. After cryotransferring and freeze-drying in situ, samples were re-cooled to liquid nitrogen temperature and low-dose (about 10³ e/nm²) digital dark-field images were obtained with single-electron sensitivity using a beam energy of approximately 100 keV and a probe current of approximately 5 pA. These maps provided a means of characterizing the molecular weights of the structures at low dose. The probe current was subsequently increased to about 5 nA in order to perform elemental analysis. The 320 copper atoms in a keyhole limpet haemocyanin molecule (mol.wt = 8 MDa) were detected with a sensitivity of ± 30 atoms in an acquisition time of 200 s. Phosphorus was detected in an approximately 10-nm length of single-stranded RNA contained in a tobacco mosaic virus particle (mol.wt = 130 kDa/nm) with a sensitivity of ± 25 atoms. Near single-atom sensitivity was achieved for the detection of iron in one haemoglobin molecule (mol.wt = 65 kDa, containing four Fe atoms). Such detection limits are only feasible if special processing methods are employed, as is demonstrated by the use of the second-difference acquisition technique and multiple least-squares fitting of reference spectra. Moreover, an extremely high electron dose (about 10¹⁰ e/nm²) is required resulting in mass loss that may be attributable to ‘knock-on’ radiation damage.     

Determination of polydatin by gold nanoparticle-enhanced chemiluminescence of silver nitrate and luminol

A novel flow injection method for the determination of polydatin is reported based on the inhibition of silver nitrate, luminol, and gold nanoparticles chemiluminescence. Under the optimum condition, the decrease in chemiluminescence was proportional to the concentration of polydatin from 1.0 × 10^?8 to 1.0 × 10^?5 mol/L. The detection limit was 2.1 × 10^?9 mo1/L and the relative standard deviation was 2.1% for the determination of 1.0 × 10^?6 mol/L polydatin. This method was successfully employed for the determination of polydatin in Polygonum cuspidatum roots and human urine.     

Acute effects of splint immobilization of the forearm on in vivo microcirculation and histomorphology of the human skin

BACKGROUND: Splint immobilization of the forearm is often performed in clinical practice. Previous studies investigated the effect of immobilization on bone, cartilage, muscle, and tendon, however, the acute effects on human skin microcirculation and histomorphology remains elusive. METHODS: In 12 healthy, nonsmoking individuals (aged 29.7 ± 9.1 years) a randomly selected forearm was immobilized by splinting for 72 h, whereas the other forearm served as control. In vivo Reflectance‐Mode Confocal‐Microscopy (RMCM) was performed prior (baseline value) and postimmobilization to evaluate: quantitative blood cell flow; density of functional dermal capillaries; epidermal thickness; and granular cell size. RESULTS: At 72h forearm immobilization, quantitative blood cell flow was significantly reduced (42.86 ± 3.68 cells/min) compared to the control blood flow (53.11 ± 3.68 cells/min, P < 0.05) and dermal capillaries indicates less functional density (5.73 ± 0.63 capillaries/mm²) compared to the controls (7.04 ± 0.81 capillaries/mm², P < 0.05). Histometric assessment reveals significantly thinner epidermis following immobilization compared to the control site (40.02 ± 2.91 vs. 46.64±3.09 µm, P < 0.05). Granular cell size was significantly altered at 72 h splinting (730.1 ± 42.53 µm²) compared to the control cell size at 770.2 ± 38.21 µm². Comparison of baseline values of both forearms indicate statistically insignificance (P > 0.05) for each parameter. CONCLUSION: At 72 h splint immobilization, for the first time, significant adaptive mechanisms were evaluated on human skin microcirculation and histomorphology using in vivo RMCM. These adaptations may be considered as an incipient atrophy of the human skin. Long‐term effects of immobilization including the regenerative potential should be evaluated in further RMCM studies. Microsc. Res. Tech. 77:99–103, 2014. © 2013 Wiley Periodicals, Inc.     

Construction of an Optical Sensor for Cobalt Determination Based on Methyltrioctylammonium Chloride Immobilized on a Polymer Membrane

Abstract

An optical sensor has been designed for the determination of cobalt by spectrophotometry. The sensing membrane is made by immobilizing methyltrioctylammonium chloride on a triacetylcellulose membrane. In the presence of Co(II) and thiocyanate ions, the colorless membrane changes to blue. The response time of the optode was about 7 min. The sensor can readily be regenerated with 0.02 mol L^?1 sodium oxalate solution. This optode is stable and can be stored under water for more than a month without reagent leaching. The calibration curve was linear in the range of 8.5×10^?6–1.3×10^?4 mol L^?1 of Co(II) ion with a limit of detection 5.9×10^?6 mol L^?1. The relative standard deviations for seven replicate measurements of 3.4×10^?5 and 1×10^?4 mol L^?1 of Co(II) were 1.58 and 1.10%, respectively. The sensor was successfully applied to the determination of cobalt in food samples and vitamin B₁₂ ampoule.     

Immunoregulatory effects of human amniotic mesenchymal stem cells and their exosomes on human peripheral blood mononuclear cells

Background: The immunomodulatory effects of mesenchymal stem cells (MSCs) and their exosomes have been receiving increasing attention. This study investigated the immunoregulatory effects of human amniotic mesenchymal stem cells (hAMSCs) and their exosomes on phytohemagglutinin (PHA)-induced peripheral blood mononuclear cells (PBMCs). Methods: The hAMSCs used in the experiment were identified by light microscopy and flow cytometry, and the differentiation ability of the cells was determined by Oil Red O and Alizarin Red staining. The expressions of transforming growth factor (TGF)-β, indoleamine 2,3-dioxygenase (IDO), cyclooxygenase-2 (COX-2), hepatocyte growth factor (HGF), and interleukin (IL)-6 were detected by quantitative real-time polymerase chain reaction and western blotting. PBMCs, hAMSCs, and their exosomes were collected for in vitro group culture. Then the immunoregulatory ability of hAMSCs and their exosomes were analyzed by flow cytometry and Enzyme-linked immunosorbent assay. Results: The hAMSCs and exosomes were successfully extracted from the human amniotic membrane. TGF-β, IDO, COX-2, HGF, and IL-6 were significantly expressed in hAMSCs. In vitro co-culture showed that hAMSCs promoted the proliferation of Th2 cells in PHA-induced PBMCs, while hAMSCs and exosomes inhibited the secretion of TNF-α in PHA-induced PBMCs, and promoted the secretion of IL-4 and IL-10, and hAMSCs had more significant effects than exosomes. Conclusions: hAMSCs or exosomes could exert immunoregulatory effects on PHA-induced PBMCs by affecting Th2 cell proliferation and cytokine secretion.     

Detection of sulfur in fixed and embedded rat peritoneal mast cell granules by X-ray energy dispersive spectroscopy

Purified rat peritoneal mast cells contained 3.3 × 10^?5 gm SO₄ and 2.2 × 10^?8 gm Ca/10⁶ cells. The molar ratio of S/Ca in the whole cell was 600:1. Frozen thin sections of unfixed mast cells contained only sulfur (S) in the granules when examined by X-ray energy dispersive spectroscopy (EDS). Mast cells fixed in 3% glutaraldehyde and 1.5% formaldehyde in 75% ethanol (Et/Ald) or in mixed buffered aldehydes and embedded in Epon 812 or the low viscosity resin diepoxyoctane (DEO) contained S in all granules and Ca in some of the granules measured. Neither element was found in the nucleus, cytoplasm, or resin. Isolated, Et/Ald fixed and embedded granules also contained S. The presence of Ca in the granules was artifactual in that the Ca was absorbed from water in the trough of the diamond knife and/or from the filter paper used to blot the sections dry. This phenomenon was investigated further. Sections of Et/Ald fixed and embedded mast cells were incubated with 5 × 10^?6 to 10^?2 M CaCl₂. Ca was detectable in 100% of the granules incubated at concentrations ≥ 10^?4 M and reached a constant S/Ca ratio of 2.0 at concentrations ≥ 10^?3 M. Ca was not detectable in the nucleus, cytoplasm, or resin at 10^?2 M. A plot of S versus Ca counts from the granules of cells incubated with 10^?2 M CaCl₂ was linear with a slope of 2.0 and a correlation coefficient of 0.88. Et/Ald fixed cells incubated with distilled H₂O had fewer granules containing Ca, 10%, than unincubated cells, 77%. Further, H₂O removed all Ca from Et/Ald fixed cells embedded in DEO. These studies show that S, which is present as SO₄ on the proteoglycan heparin, is readily detectable by X-ray EDS in fixed and embedded cells. An artifact of the technique is that weak anionic sites, which are most probably carboxyl groups on the proteoglycan, can bind the divalent cation Ca and cause spurious localization.     

Fabrication and Evaluation of Postcapillary Junctions Via Micro-Scale Molding for Use as Reactors and Flow Multiplexers in Capillary Electrokinetic Separations

Abstract

A new method of micro-fabrication is developed and employed in the production of low dead volume junctions for two post capillary detection schemes. The application of the junction as a post capillary reactor for peroxyoxalate cheinifuminescence detection is presented. Approximate detection limits of 2.5 x 10^-7 M and 9.1 x 10^-7 M are reported for perylene and anthracene (respectively) with a response linear over 3 orders of magnitude. A novel post capillary flow multiplexer is reported which utilizes electroosmotic flow switching to divert bands into secondary channels for off-line detection. The effect of solvent composition and reservoir geometry on electroosmotic flow switching times is described.