126 results on '"Guijt RM"'
Search Results
2. Exploring chip-capillary electrophoresis-laser-induced fluorescence field-deployable platform flexibility: Separations of fluorescent dyes by chip-based non-aqueous capillary electrophoresis
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Nuchtavorn, N, Smejkal, P, Breadmore, MC, Guijt, RM, Doble, P, Bek, F, Foret, F, Suntornsuk, L, and Macka, M
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Electrophoresis, Microchip ,Water ,Regression Analysis ,Reproducibility of Results ,Dimethyl Sulfoxide ,Acetates ,Sensitivity and Specificity ,Fluorescence ,Analytical Chemistry ,Fluorescent Dyes - Abstract
Microfluidic chip electrophoresis (chip-CE) is a separation method that is compatible with portable and on-site analysis, however, only few commercial chip-CE systems with laser-induced fluorescence (LIF) and light emitting diode (LED) fluorescence detection are available. They are established for several application tailored methods limited to specific biopolymers (DNA, RNA and proteins), and correspondingly the range of their applications has been limited. In this work we address the lack of commercially available research-type flexible chip-CE platforms by exploring the limits of using an application-tailored system equipped with chips and methods designed for DNA separations as a generic chip-CE platform - this is a very significant issue that has not been widely studied. In the investigated Agilent Bioanalyzer chip-CE system, the fixed components are the Agilent chips and the detection (LIF at 635nm and LEDIF at 470nm), while the chemistry (electrolyte) and the programming of all the high voltages are flexible. Using standard DNA chips, we show that a generic CE function of the system is easily possible and we demonstrate an extension of the applicability to non-aqueous CE (NACE). We studied the chip compatibility with organic solvents (i.e. MeOH, ACN, DMF and DMSO) and demonstrated the chip compatibility with DMSO as a non-volatile and non-hazardous solvent with satisfactory stability of migration times over 50h. The generic CE capability is illustrated with separations of fluorescent basic blue dyes methylene blue (MB), toluidine blue (TB), nile blue (NB) and brilliant cresyl blue (BC). Further, the effects of the composition of the background electrolyte (BGE) on the separation were studied, including the contents of water (0-30%) and buffer composition. In background electrolytes containing typically 80mmol/L ammonium acetate and 870mmol/L acetic acid in 100% DMSO baseline separation of the dyes were achieved in 40s. Linearity was documented in the range of 5-28μmol/L, 10-100μmol/L, 1.56-50nmol/L and 5-75nmol/L (r2 values in the range 0.974-0.999), and limit of detection (LOD) values were 90nmol/L, 1μmol/L 1.4nmol/L, and 2nmol/L for MB, TB, NB and BC, respectively. © 2013 Elsevier B.V.
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- 2013
3. Electrokinetic supercharging for on-line preconcentration of seven non-steroidal anti-inflammatory drugs in water samples.
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Dawod, MDZ, Breadmore, MC, Guijt, RM, Haddad, PR, Dawod, MDZ, Breadmore, MC, Guijt, RM, and Haddad, PR
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The development of new sensitive methods for the analysis of non-steroidal anti-inflammatory drugs (NSAIDs) in water samples is of great importance. In this work, seven NSAIDs were separated within 9 min using 15 mM sodium tetraborate (pH 9.2) containing 0.1% (w/v) hexadimethrine bromide (HDMB) and 10% (v/v) methanol. Field-amplified sample injection (FASI) was examined and found to improve the detection limits by 200-fold providing detection limits of 0.6–2.0 μg/L, but these are insufficient for the determination of NSAIDs as environmental pollutants in water samples. To improve the sensitivity further, electrokinetic supercharging (EKS) was examined. The optimum EKS method involved hydrodynamic injection leading electrolyte (100 mM NaCl, 30 s, 50 mbar), electrokinetic injection of the sample (200 s, −10 kV) and finally injection of the terminating electrolyte (100 mM 2-(cyclohexylamino) ethanesulphonic acid, CHES, 40 s, 50 mbar). With this method, the sensitivity was improved by 2400-fold giving detection limits of 50–180 ng/L. The developed method was validated and then applied to the analysis of wastewater samples from a local sewage treatment plant. The detection limits were found to increase by approximately 10-fold, however, this is still lower than levels previously found in wastewater samples from European and Mediterranean cities. The proposed method has the advantage of simplicity and achieving sensitivity through high-preconcentration power without the use of off-line chromatographic sample cleanup.
4. New approaches for fabrication of microfluidic capillary electrophoresis devices with on-chip conductivity detection
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Guijt, RM, Baltussen, E, van der Steen, G, Schasfoort, RBM, Schlautmann, S, Billiet, HAH, Frank, J, van Dedem, GWK, van den Berg, A, Guijt, RM, Baltussen, E, van der Steen, G, Schasfoort, RBM, Schlautmann, S, Billiet, HAH, Frank, J, van Dedem, GWK, and van den Berg, A
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In practice, microfluidic systems are based on the principles of capillary electrophoresis (CE), for a large part due to the simplicity of electroosmotic pumping. In this contribution, a universal conductivity detector is presented that allows detection of charged species down to the M level. Additionally, powderblasting is presented as a novel technique for direct etching of microfluidic networks. This method allows creation of features down to 50 μM with a total processing time (design to device) of less than one day. The performance of powderblasted devices with integrated conductivity detection is illustrated by the separation of lithium, sodium, and potassium ions and that of fumaric, malic, and citric acid.
5. Considerations on contactless conductivity detection in capillary electrophoresis
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Baltussen, E, Guijt, RM, van der Steen, G, Laugere, F, Baltussen, S, van Dedem, GWK, Baltussen, E, Guijt, RM, van der Steen, G, Laugere, F, Baltussen, S, and van Dedem, GWK
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Nearly all analyses by capillary electrophoresis (CE) are performed using optical detection, utilizing either absorbance or (laser-induced) fluorescence. Though adequate for many analytical problems, in a large number of cases, e.g., involving non-UV-absorbing compounds, these optical detection methods fall short. Indirect optical detection can then still provide an acceptable means of detection, however, with a strongly reduced sensitivity. During the past few years, contactless conductivity detection (CCD) has been presented as a valuable extension to optical detection techniques. It has been demonstrated that with CCD detection limits comparable, or even superior, to (indirect) optical detection can be obtained. Additionally, construction of the CCD around the CE capillary is straightforward and robust operation is easily obtained. Unfortunately, in the literature a large variety of designs and operating conditions for CCD were described. In this contribution, several important parameters of CCD are identified and their influence on, e.g., detectability and peak shape is described. An optimized setup based on a well-defined detection cell with three detection electrodes is presented. Additionally, simple and commercially available read-out electronics are described. The performance of the CCD-CE system was demonstrated for the analysis of peptides. Detection limits at the μM level were obtained in combination with good peak shapes and an overall good performance and stability.
6. Capillary electrophoresis with on-chip four-electrode capacitively coupled conductivity detection for application in bioanalysis
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Guijt, RM, Baltussen, E, van der Steen, G, Frank, J, Billiet, HAH, Schalkhammer, T, Laugere, F, Vellekoop, MJ, Berthold, A, Sarro, PM, van Dedem, GWK, Guijt, RM, Baltussen, E, van der Steen, G, Frank, J, Billiet, HAH, Schalkhammer, T, Laugere, F, Vellekoop, MJ, Berthold, A, Sarro, PM, and van Dedem, GWK
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Microchip capillary electrophoresis (CE) with integrated four-electrode capacitively coupled conductivity detection is presented. Conductivity detection is a universal detection technique that is relatively independent on the detection pathlength and, especially important for chip-based analysis, is compatible with miniaturization and on-chip integration. The glass microchip structure consists of a 6 cm etched channel (20 μm × 70 μm cross section) with silicon nitride covered walls. In the channel, a 30 nm thick silicon carbide layer covers the electrodes to enable capacitive coupling with the liquid inside the channel as well as to prevent interference of the applied separation field. The detector response was found to be linear over the concentration range from 20 μM up to 2 mM. Detection limits were at the low μm level. Separation of two short peptides with a pl of respectively 5.38 and 4.87 at the 1 mM level demonstrates the applicability for biochemical analysis. At a relatively low separation field strength (50 V/cm) plate numbers in the order of 3500 were achieved. Results obtained with the microdevice compared well with those obtained in a bench scale CE instrument using UV detection under similar conditions.
7. Indirect electro-osmotic pumping
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Guijt, RM, Lichtenberg, J, de Rooij, NIF, Verpoorte, E, Baltussen, E, van Dedem, GWK, Guijt, RM, Lichtenberg, J, de Rooij, NIF, Verpoorte, E, Baltussen, E, and van Dedem, GWK
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The manipulation of liquids within a microcapillary network remains a considerable challenge in the development of miniaturized total chemical analysis systems (μTAS). Fluid manipulation can be achieved using (micro) mechanical pumps connected or integrated into the device, and by using an electric field (E) for generation of electro-osmotic flow (EOF). For glass microdevices, electro-osmotic pumping (EOP) is most attractive, since no moving parts and/or valves are required. In its simplest embodiment, EOP in microfluidic devices involves imposing an E along the full length of the channel by immersing electrodes into open solution reservoirs situated at both ends of the channel. Electrolytically generated gases at the electrodes drift to the surface of the solution reservoirs and escape into the air. In more complex situations, however, EOP in a subsection of a microchannel may be required. For sampling, for example, from brain tissue in living organisms, the presence of electrodes in the ‘sample reservoir’ (i.e., the brain), and thus outside the microdevice is undesirable, since potentials applied to external electrodes interfere with the sampling environment. In these cases, electrodes need to be integrated into the microfluidic device. The use of electrodes in a microchannel, however, is not trivial. Electrolytic gases get caught in the sealed microchannel and hence effectively interrupt the electric field, and thus fluid movement. A number of approaches to avoid bubble formation during spatially localized application of voltages in microfluidic networks have been reported. In one example, a 1-mm-thick poly(dimethylsiloxane) (PDMS) substrate containing the microchannel was sealed with a glass cover plate containing the electrodes.1 Electrolytic gases formed at the electrodes dissipated through the highly gas-permeable PDMS film into the air. An alternative method for application of the electric field is the use of a conducting barrier between the electrodes and t
8. On-chip contactless four-electrode conductivity detection for capillary electrophoresis devices
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Laugere, F, Guijt, RM, Bastemeijer, J, van der Steen, G, Berthold, A, Baltussen, E, Sarro, PM, van Dedem, GWK, Vellekoop, MJ, Bossche, Aé, Laugere, F, Guijt, RM, Bastemeijer, J, van der Steen, G, Berthold, A, Baltussen, E, Sarro, PM, van Dedem, GWK, Vellekoop, MJ, and Bossche, Aé
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In this contribution, a capillary electrophoresis microdevice with an integrated on-chip contactless four-electrode conductivity detector is presented. A 6-cm-long, 70-μm-wide, and 20-μm-deep channel was etched in a glass substrate that was bonded to a second glass substrate in order to form a sealed channel. Four contactless electrodes (metal electrodes covered by 30-nm silicon carbide) were deposited and patterned on the second glass substrate for on-chip conductivity detection. Contactless conductivity detection was performed in either a two- or a four-electrode configuration. Experimental results confirmed the improved characteristics of the four-electrode configuration over the classical two-electrode detection setup. The four-electrode configuration allows for sensitive detection for varying carrier-electrolyte background conductivity without the need for adjustment of the measurement frequency. Reproducible electrophoretic separations of three inorganic cations (K+, Na+, Li+) and six organic acids are presented. Detection as low as 5 μM for potassium was demonstrated.
9. Internal electrolyte temperatures for polymer and fused-silica capillaries used in capillary electrophoresis
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Evenhuis, CJ, Guijt, RM, Macka, M, Marriott, PJ, Haddad, PR, Evenhuis, CJ, Guijt, RM, Macka, M, Marriott, PJ, and Haddad, PR
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Polymers are important as materials for manufacturing microfluidic devices for electro-driven separations, in which Joule heating is an unavoidable phenomenon. Heating effects were investigated in polymer capillaries using a CE setup. This study is the first step toward the longer-term objective of the study of heating effects occurring in polymeric microfluidic devices. The thermal conductivity of polymers is much smaller than that of fused silica (FS), resulting in less efficient dissipation of heat in polymeric capillaries. This study used conductance measurements as a temperature probe to determine the mean electrolyte temperatures in CE capillaries of different materials. Values for mean electrolyte temperatures in capillaries made of New Generation FluoroPolymer (NGFP), poly-(methylmethacrylate) (PMMA), and poly(ether ether ketone) (PEEK) capillaries were compared with those obtained for FS capillaries. Extrapolation of plots of conductance versus power per unit length (P/L) to zero power was used to obtain conductance values free of Joule heating effects. The ratio of the measured conductance values at different power levels to the conductance at zero power was used to determine the mean temperature of the electrolyte. For each type of capillary material, it was found that the average increase in the mean temperature of the electrolyte (ΔTMean) was directly proportional to P/L and inversely proportional to the thermal conductivity (λ) of the capillary material. At 7.5 W/m, values for ΔTMean for NGFP, PMMA, and PEEK were determined to be 36.6, 33.8, and 30.7°C, respectively. Under identical conditions, ΔTMean for FS capillaries was 20.4°C.
10. Fabrication of a glass-implemented microcapillary electrophoresis device with integrated contactless conductivity detection
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Berthold, A, Laugere, F, Schellevis, H, de Boer, CR, Laros, M, Guijt, RM, Sarro, PM, Vellekoop, MJ, Berthold, A, Laugere, F, Schellevis, H, de Boer, CR, Laros, M, Guijt, RM, Sarro, PM, and Vellekoop, MJ
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Glass microdevices for capillary electrophoresis (CE) gained a lot of interest in the development of micrototal analysis systems (μTAS). The fabrication of a μJAS requires integration of sampling, chemical separation and detection systems into a microdevice. The integration of a detection system into a microchannel, however, is hampered by the lack of suitable microfabrication technology. Here, a microfabrication method for integration of insulated microelectrodes inside a leakage-free microchannel in glass is presented. A combination of newly developed technological approaches, such as low-temperature glass-to-glass anodic bonding, channel etching, fabrication of buried metal interconnects, and deposition of thin plasma-enhanced chemical vapour deposition (PECVD) silicon carbide layers, enables the fabrication of a CE microdevice with an integrated contactless conductivity detector. The fabrication method of this CE microdevice with integrated contactless conductivity detector is described in detail. Standard CE separations of three inorganic cations in concentrations down to 5 μm show the viability of the new μCE system.
11. Recent advances in affinity capillary electrophoresis
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Guijt, RM, Frank, J, van Dedem, GWK, Baltussen, E, Guijt, RM, Frank, J, van Dedem, GWK, and Baltussen, E
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Use of the specificity of (bio)interactions can effectively overcome the selectivity limitation faced in capillary electrophoresis (CE), and the resulting technique usually is referred to as affinity capillary electrophoresis (ACE). Despite the high selectivity of ACE, several important problems still need to be addressed. A major issue in all CE separations, including ACE, is the concentration detection limit. Using UV detection, this is usually in the order of 10 - 6 M whereas laser-induced fluorescence (LIF) detection can provide detection limits down to the sub-10 - 10 M range. However, a marked disadvantage of LIF is that labeling of the analytes is usually required, which might change the interaction behavior of the solutes under investigation. Additionally, labeling reactions at sub-10 - 10 M concentration levels are certainly not trivial and often difficult to perform quantitatively. Alternative and universal detection approaches, particularly mass spectrometric (MS) detection, look very promising but (A) CE-MS techniques are still far from routine application. Important future progress in sensitive detection strategies is likely to increase the use of ACE in the future.
12. Use of bioaffinity interactions in electrokinetically controlled assays on microfabricated devices
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Guijt, RM, Baltussen, E, van Dedem, GWK, Guijt, RM, Baltussen, E, and van Dedem, GWK
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In this contribution, the role of bioaffinity interactions on electrokinetically controlled microfabricated devices is reviewed. Interesting applications reported in the literature include enzymatic assays, where enzyme and enzyme inhibition kinetics were studied, often in combination with electrophoretic separation. Attention is paid towards developments that could lead to implementation of electrokinetically controlled microdevices in high-throughput screening. Furthermore, enzyme-facilitated detection in combination with electrophoretic separation on microdevices is discussed. Various types of immunoassays have been implemented on the microchip format. The selectivity of antibody-antigen interaction has been exploited for the detection of analytes in complex sample matrices as required, for example, in clinical chemistry. Binding kinetics as well as stoichiometry were studied in chip-based assays. Automated mixing protocols as well as the demonstration of a parallel immunoassay allow implementation of microdevices in high-throughput screening. Furthermore, demonstration of immunoassays on cheap polymeric microdevices opens the way towards the fabrication of disposable devices, a requirement for commercialization and therefore for application in routine analyses.
13. Identification of homemade inorganic explosives by ion chromatographic analysis of post-blast residues
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Johns, CA, Shellie, RA, Potter, OG, O'Reilly, JW, Hutchinson, JP, Guijt, RM, Breadmore, MC, Hilder, EF, Dicinoski, GW, Haddad, PR, Johns, CA, Shellie, RA, Potter, OG, O'Reilly, JW, Hutchinson, JP, Guijt, RM, Breadmore, MC, Hilder, EF, Dicinoski, GW, and Haddad, PR
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Anions and cations of interest for the post-blast identification of homemade inorganic explosives were separated and detected by ion chromatographic (IC) methods. The ionic analytes used for identification of explosives in this study comprised 18 anions (acetate, benzoate, bromate, carbonate, chlorate, chloride, chlorite, chromate, cyanate, fluoride, formate, nitrate, nitrite, perchlorate, phosphate, sulfate, thiocyanate and thiosulfate) and 12 cations (ammonium, barium(II), calcium(II), chromium(III), ethylammonium, magnesium(II), manganese(II), methylammonium, potassium(I), sodium(I), strontium(II), and zinc(II)). Two IC separations are presented, using suppressed IC on a Dionex AS20 column with potassium hydroxide as eluent for anions, and non-suppressed IC for cations using a Dionex SCS 1 column with oxalic acid/acetonitrile as eluent. Conductivity detection was used in both cases. Detection limits for anions were in the range 2–27.4 ppb, and for cations were in the range 13–115 ppb. These methods allowed the explosive residue ions to be identified and separated from background ions likely to be present in the environment. Linearity (over a calibration range of 0.05–50 ppm) was evaluated for both methods, with r2 values ranging from 0.9889 to 1.000. Reproducibility over 10 consecutive injections of a 5 ppm standard ranged from 0.01 to 0.22% relative standard deviation (RSD) for retention time and 0.29 to 2.16%RSD for peak area. The anion and cation separations were performed simultaneously by using two Dionex ICS-2000 chromatographs served by a single autoinjector. The efficacy of the developed methods was demonstrated by analysis of residue samples taken from witness plates and soils collected following the controlled detonation of a series of different inorganic homemade explosives. The results obtained were also confirmed by parallel analysis of the same samples by capillary electrophoresis (CE) with excellent agreement being obtained.
14. Miniaturized analytical assays in biotechnology
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Guijt, RM, Moerman, R, Kroon, A, van Dedem, GWK, van den Doel, R, van Vliet, L, Young, IT, Laugere, F, Bossche, Aé, Sarro, PM, Guijt, RM, Moerman, R, Kroon, A, van Dedem, GWK, van den Doel, R, van Vliet, L, Young, IT, Laugere, F, Bossche, Aé, and Sarro, PM
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Biotechnology today is a well-established paradigm in many areas of human endeavor, such as the pharmaceutical industry, agriculture, management of the environment and many others. Meanwhile, biology is undergoing a spectacular transition: whereas systematic biology was replaced gradually by molecular biology, the latter is rapidly being transformed into a new systematic era in which entire genomes are being charted by ever more sophisticated analytical techniques. In the wake of this onslaught of data, new fields are germinating, such as bioinformatics in an attempt to find answers to fundamental questions, answers that may be hidden in the massive amounts of data already available today.
15. Identification of inorganic improvised explosive devices by analysis of postblast residues using portable capillary electrophoresis instrumentation and indirect photometric detection with a light-emitting diode
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Hutchinson, JP, Evenhuis, CJ, Johns, CA, Kazarian, AA, Breadmore, MC, Macka, M, Hilder, EF, Guijt, RM, Dicinoski, GW, Haddad, PR, Hutchinson, JP, Evenhuis, CJ, Johns, CA, Kazarian, AA, Breadmore, MC, Macka, M, Hilder, EF, Guijt, RM, Dicinoski, GW, and Haddad, PR
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A commercial portable capillary electrophoresis (CE) instrument has been used to separate inorganic anions and cations found in postblast residues from improvised explosive devices (IEDs) of the type used frequently in terrorism attacks. The purpose of this analysis was to identify the type of explosive used. The CE instrument was modified for use with an in-house miniaturized light-emitting diode (LED) detector to enable sensitive indirect photometric detection to be employed for the detection of 15 anions (acetate, benzoate, carbonate, chlorate, chloride, chlorite, cyanate, fluoride, nitrate, nitrite, perchlorate, phosphate, sulfate, thiocyanate, thiosulfate) and 12 cations (ammonium, monomethylammonium, ethylammonium, potassium, sodium, barium, strontium, magnesium, manganese, calcium, zinc, lead) as the target analytes. These ions are known to be present in postblast residues from inorganic IEDs constructed from ammonium nitrate/fuel oil mixtures, black powder, and chlorate/perchlorate/sugar mixtures. For the analysis of cations, a blue LED (470 nm) was used in conjunction with the highly absorbing cationic dye, chrysoidine (absorption maximum at 453 nm). A nonaqueous background electrolyte comprising 10 mM chrysoidine in methanol was found to give greatly improved baseline stability in comparison to aqueous electrolytes due to the increased solubility of chrysoidine and its decreased adsorption onto the capillary wall. Glacial acetic acid (0.7% v/v) was added to ensure chrysoidine was protonated and to enhance separation selectivity by means of complexation with transition metal ions. The 12 target cations were separated in less than 9.5 min with detection limits of 0.11-2.30 mg/L (calculated at a signal-to-noise ratio of 3). The anions separation system utilized a UV LED (370 nm) in conjunction with an aqueous chromate electrolyte (absorption maximum at 371 nm) consisting of 10 mM chromium(VI) oxide and 10 mM sodium chromate, buffered with 40 mM tris(hydroxyme
16. Determination of inorganic ions using microfluidic devices
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Evenhuis, CJ, Guijt, RM, Macka, M, Haddad, PR, Evenhuis, CJ, Guijt, RM, Macka, M, and Haddad, PR
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The separation and detection of inorganic ions on microfluidic devices has received little attention since the lab-on-a-chip concept has revolutionised the field of electrokinetically driven analysis. This review presents a summary and discussion of the published literature on inorganic analysis using microfluidic devices and includes sections on electromigration separation methods, namely isotachophoresis (ITP), capillary electrophoresis (CE), and hyphenated ITP-CE, together with a brief account of flow injection analysis. The review concludes with the authors' perspective on future directions for inorganic analysis on microfluidic devices.
17. Conductivity detection for conventional and miniaturised capillary electrophoresis systems
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Guijt, RM, Evenhuis, CJ, Macka, M, Haddad, PR, Guijt, RM, Evenhuis, CJ, Macka, M, and Haddad, PR
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Since the introduction of capillary electrophoresis (CE), conductivity detection has been an attractive means of detection. No additional chemical properties are required for detection, and no loss in sensitivity is expected when miniaturising the detector to scale with narrow-bore capillaries or even to the microchip format. Integration of conductivity and CE, however, involves a challenging combination of engineering issues. In conductivity detection the resistance of the solution is most frequently measured in an alternating current (AC) circuit. The influence of capacitors both in series and in parallel with the solution resistance should be minimised during conductivity measurements. For contact conductivity measurements, the positioning and alignment of the detection electrodes is crucial. A contact conductivity detector for CE has been commercially available, but was withdrawn from the market. Microfabrication technology enables integration and precise alignment of electrodes, resulting in the popularity of conductivity detection in microfluidic devices. In contactless conductivity detection, the alignment of the electrodes with respect to the capillary is less crucial. Contactless conductivity detection (CCD) was introduced in capillary CE, and similar electronics have been applied for CCD using planar electrodes in microfluidic devices. A contactless conductivity detector for capillaries has been commercialised recently. In this review, different approaches towards conductivity detection in capillaries and chip-based CE are discussed. In contrast to previous reviews, the focus of the present review is on the technological developments and challenges in conductivity detection in CE.
18. Variation of zeta-potential with temperature in fused-silica capillaries used for capillary electrophoresis
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Evenhuis, CJ, Guijt, RM, Macka, M, Marriott, PJ, Haddad, PR, Evenhuis, CJ, Guijt, RM, Macka, M, Marriott, PJ, and Haddad, PR
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The temperature variation of electroosmotic mobility corrected for the effects of Joule heating was employed to investigate the variation of the zeta-potential with temperature in fused-silica capillaries. Experimentally determined values for zeta increased at 0.39% per °C, a rate that is about four to five times smaller than reported previously. Experimentally determined values of zeta were directly proportional to the absolute temperature although values were also influenced slightly by changes to the dielectric constant. It was found that the effective charge density at the inner surface of the capillary was independent of temperature.
19. Reliable electrophoretic mobilities free from joule heating effects using Capillary Electrophoresis
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Evenhuis, CJ, Hruska, J, Guijt, RM, Macka, M, Gaš, B, Marriott, PJ, Haddad, PR, Evenhuis, CJ, Hruska, J, Guijt, RM, Macka, M, Gaš, B, Marriott, PJ, and Haddad, PR
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Ionic electrophoretic mobilities determined by means of CE experiments are sometimes different when compared to generally accepted values based on limiting ionic conductance measurements. While the effect of ionic strength on electrophoretic mobility has been long understood, the increase in the mobility that results from Joule heating (the resistive heating that occurs when a current passes through an electrolyte) has been largely overlooked. In this work, a simple method for obtaining reliable and reproducible values of electrophoretic mobility is described. The electrophoretic mobility is measured over a range of driving powers and the extrapolation to zero power dissipation is employed to eliminate the effect of Joule heating. These extrapolated values of electrophoretic mobility can then be used to calculate limiting ionic mobilities by making a correction for ionic strength; this somewhat complicated calculation is conveniently performed by using the freeware program PeakMaster 5. These straightforward procedures improve the agreement between experimentally determined and literature values of limiting ionic mobility by at least one order of magnitude. Using Tris-chromate BGE with a value of conductivity 0.34 S/m and ionic strength 59 mM at a modest dissipated power per unit length of 2.0 W/m, values of mobility for inorganic anions were increased by an average of 12.6% relative to their values free from the effects of Joule heating. These increases were accompanied by a reduction in mobilities due to the ionic strength effect, which was 11% for univalent and 28% for divalent inorganic ions compared to their limiting ionic mobilities. Additionally, it was possible to determine the limiting ionic mobility for a number of aromatic anions by using PeakMaster 5 to perform an ionic strength correction. A major significance of this work is in being able to use CE to obtain reliable and accurate values of electrophoretic mobilities with all its benefits, including und
20. Temperature profiles and heat dissipation in capillary electrophoresis
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Evenhuis, CJ, Guijt, RM, Macka, M, Marriott, PJ, Haddad, PR, Evenhuis, CJ, Guijt, RM, Macka, M, Marriott, PJ, and Haddad, PR
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While temperature control is usually employed in capillary electrophoresis (CE) to aid heat dissipation and provide acceptable precision, internal electrolyte temperatures are almost never measured. In principle, this limits the accuracy, repeatability, and method robustness. This work presents a fundamental study that combines the development of new equations characterizing temperature profiles in CE with a new method of temperature determination. New equations were derived from first principles relating the mean, axial, and inner wall electrolyte temperatures (TMean, TAxis, TWall). TMean was shown to occur at a distance 1/3 times the internal radius of the capillary from the center of the capillary and to be a weighted average of 2/3TAxis and 1/3TWall. Conductance (G) and electroosmotic mobility (EOF) can be used to determine TMean and TWall, respectively. Extrapolation of curves of EOF versus power per unit length (P/L) at different temperatures was used to calibrate the variation of EOF with temperature (T), free from Joule heating effects. EOF increased at 2.22%/C. The experimentally determined temperatures using EOF agreed to within 0.2 C with those determined using G. The accuracy of G measurements was confirmed independently by measuring the electrical conductivity () of the bulk electrolyte over a range of temperatures and by calculating the variation of G with T from the Debye-Hückel-Onsager equation. TMean was found to be up to 20 C higher than the external temperature under typical conditions using active air-cooling and a 74.0-m-internal diameter (di) fused-silica capillary. A combination of experimentally determined and calculated temperatures enables a complete temperature profile for a fused-silica capillary to be drawn and the thickness of the stationary air layer to be determined. As an example, at P/L = 1.00 Wm-1, the determined radial temperature difference across the electrolyte was 0.14 C; the temperature difference across the fused-silica wall
21. Chemical and physical processes for integrated temperature control in microfluidic devices
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Guijt, RM, Dodge, A, van Dedem, GWK, de Rooij, NF, Verpoorte, E, Guijt, RM, Dodge, A, van Dedem, GWK, de Rooij, NF, and Verpoorte, E
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Microfluidic devices are a promising new tool for studying and optimizing (bio)chemical reactions and analyses. Many (bio)chemical reactions require accurate temperature control, such as for example thermocycling for PCR. Here, a new integrated temperature control system for microfluidic devices is presented, using chemical and physical processes to locally regulate temperature. In demonstration experiments, the evaporation of acetone was used as an endothermic process to cool a microchannel. Additionally, heating of a microchannel was achieved by dissolution of concentrated sulfuric acid in water as an exothermic process. Localization of the contact area of two flows in a microfluidic channel allows control of the position and the magnitude of the thermal effect.
22. Capillary Microreactor Catalysis Utilising Solid Organic Macroporous Monolith Supports
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Bolton, KF, Canty, AJ, Deverell, JA, Guijt, RM, Hilder, EF, Rodemann, T, Smith, JA, Goemann, A, Bolton, KF, Canty, AJ, Deverell, JA, Guijt, RM, Hilder, EF, Rodemann, T, Smith, JA, and Goemann, A
23. Macroporous monolith supports for continuous flow capillary microreactors
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Bolton, KF, Canty, AJ, Deverell, JA, Guijt, RM, Hilder, EF, Rodemann, T, Smith, JA, Bolton, KF, Canty, AJ, Deverell, JA, Guijt, RM, Hilder, EF, Rodemann, T, and Smith, JA
- Abstract
A solid macroporous monolith is shown to be a suitable substrate for anchoring a palladium complex to obtain a continuous porous material suitable for conducting flow-through catalysis in capillary microreactors. The monolith for this study was GMA-co-EDMA [poly(glycidyl methacrylate-co-ethylene dimethacrylate)] in fused silica capillaries (250 micrometre internal diameter). Immobilisation of ligand was achieved by passing a solution of 5-hydroxy-1,10-phenanthroline, and palladium(II) was added. Suzuki-Miyaura catalysis was studied. The excellent flow-through properties, firm attachment to capillary walls, and robustness of the capillary systems for catalysis demonstrated after continuous reaction for four days indicate that this approach is suitable for further development of capillary-based microreactors that are suitable for continuous flow processes and parallel synthesis.
24. Capillary Microreactor Catalysis Utilising Solid Organic Macroporous Monolith Supports
- Author
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Bolton, KF, Canty, AJ, Deverell, JA, Guijt, RM, Hilder, EF, Rodemann, T, Smith, JA, Goemann, A, Bolton, KF, Canty, AJ, Deverell, JA, Guijt, RM, Hilder, EF, Rodemann, T, Smith, JA, and Goemann, A
25. Nanofluidic trap for DNA extraction from biological samples
- Author
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Shallan, AI, Guijt, RM, Breadmore, MC, and 19th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2015 Gyeongju, South Korea 25-29 October 2015
- Subjects
controlled dielectric breakdown ,electrokinetic trapping ,sample preparation ,SsDNA - Abstract
A new device seamlessly integrating various electrokinetic processes enables the analysis of short ssDNA (20 bases) directly from whole blood. An electrokinetic trap was integrated to achieve extraction, concentration, and desalting of ssDNA in 200 s followed by a 90 s electrophoretic separation. The trap consists of two sequentially coupled nanojunctions with different pore size and exhibits selective ion transport properties. Depletion forces near the nanojunctions and the apparent electromigration were tuned to enable the trapping and subsequent analysis of fluorescein labeled-ssDNA. The quantitation limit (S/N ratio = 10) was 12.5 nM using 40 μL of spiked whole blood. Refereed/Peer-reviewed
- Published
- 2015
26. Electrokinetic traps for integrated biological sample analysis
- Author
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Shallan, AI, Guijt, RM, Breadmore, MC, and 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences San Antonio, USA 26-30 October 2014
- Subjects
sample preparation, polydimethylsiloxane breakdown, electrophoresis - Abstract
A new device integrating diverse electrokinetic steps is presented to extract and enrich analytes directly from blood followed by electrophoretic separation. Two sequentially coupled nanojunctions with different pore size formed a trap with selective ion transport properties. The balance between depletion forces near the nanojunctions and/or apparent electromigration can be tuned for analysis of proteins, DNA, or pharmaceuticals. Here, we demonstrate the analysis of ampicillin from blood within 5 min. The obtained linear range of 2.5-20 μg/mL covers the therapeutically relevant concentration for treating sepsis. Refereed/Peer-reviewed
- Published
- 2014
27. Method for lysis and paper-based elution-free DNA extraction with colourimetric isothermal amplification.
- Author
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Lee SM, Doeven EH, Yuan D, and Guijt RM
- Subjects
- Colorimetry methods, DNA, Solid Phase Extraction methods, Polyethylene Glycols chemistry, DNA, Bacterial isolation & purification, DNA, Bacterial genetics, Molecular Diagnostic Techniques, Nucleic Acid Amplification Techniques methods, Paper
- Abstract
Nucleic acid amplification testing has great potential for point-of-need diagnostic testing with high detection sensitivity and specificity. Current sample preparation is limited by a tedious workflow requiring multiple steps, reagents and instrumentation, hampering nucleic acid testing at point of need. In this study, we present the use of mixed cellulose ester (MCE) paper for DNA binding by ionic interaction under molecular crowding conditions and fluid transport by wicking. The poly(ethylene) glycol-based (PEG) reagent simultaneously provides the high pH for alkaline lysis and crowding effects for ionic binding of the DNA under high salt conditions. In this study, we introduce Paper-based Abridged Solid-Phase Extraction with Alkaline Poly(ethylene) Glycol Lysis (PASAP). The anionic mixed cellulose ester (MCE) paper is used as solid phase and allows for fluid transport by wicking, eliminating the need for pipetting skills and the use of a magnet to retain beads. Following the release of DNA from the cells due to the lytic activity of the PASAP solution, the DNA binds to the anionic surface of the MCE paper, concentrating at the bottom while the sample matrix is transported towards the top by wicking. The paper was washed by dipping it in 40% isopropanol for 10 s. After air-drying for 30 s, the bottom section of the paper (3 mm × 4 mm) was snapped off using the cap of a PCR tube and immersed in the colourimetric loop-mediated isothermal amplification (cLAMP) solution for direct amplification and colourimetric detection. The total sample processing was completed in 15 min and ready for amplification. cLAMP enabled the detection of 10
2 CFU/mL of Escherichia coli (E. coli) from culture media and the detection of E. coli in milk < 103 CFU/mL (10 CFU) after incubation at 68 °C for 60 min, demonstrating applicability of the method to complex biological samples., (© 2024. The Author(s).)- Published
- 2024
- Full Text
- View/download PDF
28. 3D printed porous membrane integrated devices to study the chemoattractant induced behavioural response of aquatic organisms.
- Author
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Kalathil Balakrishnan H, Schultz AG, Lee SM, Alexander R, Dumée LF, Doeven EH, Yuan D, and Guijt RM
- Subjects
- Humans, Animals, Porosity, Tissue Scaffolds, Printing, Three-Dimensional, Aquatic Organisms, Zebrafish
- Abstract
Biological models with genetic similarities to humans are used for exploratory research to develop behavioral screening tools and understand sensory-motor interactions. Their small, often mm-sized appearance raises challenges in the straightforward quantification of their subtle behavioral responses and calls for new, customisable research tools. 3D printing provides an attractive approach for the manufacture of custom designs at low cost; however, challenges remain in the integration of functional materials like porous membranes. Nanoporous membranes have been integrated with resin exchange using purpose-designed resins by digital light projection 3D printing to yield functionally integrated devices using a simple, economical and semi-automated process. Here, the impact of the layer thickness and layer number on the porous properties - parameters unique for 3D printing - are investigated, showing decreases in mean pore diameter and porosity with increasing layer height and layer number. From the same resin formulation, materials with average pore size between 200 and 600 nm and porosity between 45% and 61% were printed. Membrane-integrated devices were used to study the chemoattractant induced behavioural response of zebrafish embryos and planarians, both demonstrating a predominant behavioral response towards the chemoattractant, spending >85% of experiment time in the attractant side of the observation chamber. The presented 3D printing method can be used for printing custom designed membrane-integrated devices using affordable 3D printers and enable fine-tuning of porous properties through adjustment of layer height and number. This accessible approach is expected to be adopted for applications including behavioural studies, early-stage pre-clinical drug discovery and (environmental) toxicology.
- Published
- 2024
- Full Text
- View/download PDF
29. Abridged solid-phase extraction with alkaline Poly(ethylene) glycol lysis (ASAP) for direct DNA amplification.
- Author
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Lee SM, Nai YH, Doeven EH, Balakrishnan HK, Yuan D, and Guijt RM
- Subjects
- Solid Phase Extraction, Ethylenes, Glycols, DNA genetics, Nucleic Acid Amplification Techniques
- Abstract
Complexity of sample preparation decelerate the development of sample-in-answer-out devices for point-of-need nucleic acid amplification testing. Here, we present the consolidation of alkaline poly(ethylene) glycol-based lysis and solid-phase extraction for rapid and simple sample preparation compatible with direct on-bead amplification. Simultaneous cell lysis and binding of DNA were achieved using an optimised reagent comprising 15% PEG8000, 0.5 M NaCl, and 3.5 mM KOH. This was combined with direct, on-bead amplification using 1.5 μg beads per 20 μL PCR reaction mix. The novel single reagent, 5-min method improved the detection limit by 10 and 100-fold compared with commercial DNA extraction kits and the original alkaline PEG lysis method, respectively. The sensitivity can be further enhanced by one amplification cycle with an ethanol wash or by extending the incubation to 10 min before collecting the magnetic particles. Both methods successfully detected a single copy of Escherichia coli DNA. In biological fluids (saliva, sweat, and urine), the 5-min method was delayed by about one cycle compared to the 15-min method. The proposed methods are attractive for incorporation in the workflow for point-of-need testing of biological samples by providing a practical and chemical method for simple alternative DNA sample preparation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)
- Published
- 2024
- Full Text
- View/download PDF
30. Chemical Trends in Sample Preparation for Nucleic Acid Amplification Testing (NAAT): A Review.
- Author
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Lee SM, Balakrishnan HK, Doeven EH, Yuan D, and Guijt RM
- Subjects
- Humans, Reproducibility of Results, Ecosystem, Nucleic Acid Amplification Techniques methods, Nucleic Acids, Communicable Diseases diagnosis
- Abstract
Nucleic acid amplification testing facilitates the detection of disease through specific genomic sequences and is attractive for point-of-need testing (PONT); in particular, the early detection of microorganisms can alert early response systems to protect the public and ecosystems from widespread outbreaks of biological threats, including infectious diseases. Prior to nucleic acid amplification and detection, extensive sample preparation techniques are required to free nucleic acids and extract them from the sample matrix. Sample preparation is critical to maximize the sensitivity and reliability of testing. As the enzymatic amplification reactions can be sensitive to inhibitors from the sample, as well as from chemicals used for lysis and extraction, avoiding inhibition is a significant challenge, particularly when minimising liquid handling steps is also desirable for the translation of the assay to a portable format for PONT. The reagents used in sample preparation for nucleic acid testing, covering lysis and NA extraction (binding, washing, and elution), are reviewed with a focus on their suitability for use in PONT.
- Published
- 2023
- Full Text
- View/download PDF
31. Development of a pumpless acoustofluidic device for rapid food pathogen detection.
- Author
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Nilghaz A, Lee SM, Su H, Yuan D, Tian J, Guijt RM, and Wang X
- Subjects
- Microfluidics methods, Acoustics, Fruit, Transducers, Escherichia coli, Microfluidic Analytical Techniques
- Abstract
Mixing, homogenization, separation, and filtration are crucial processes in miniaturized analytical systems employed for in-vitro biological, environmental, and food analysis. However, in microfluidic systems achieving homogenization becomes more challenging due to the laminar flow conditions, which lack the turbulent flows typically used for mixing in traditional analytical systems. Here, we introduce an acoustofluidic platform that leverages an acoustic transducer to generate microvortex streaming, enabling effective homogenizing of food samples. To reduce reliance on external equipment, tubing, and pump, which is desirable for Point-of-Need testing, our pumpless platform employs a hydrophilic yarn capable of continuous wicking for sample perfusion. Following the homogenization process, the platform incorporates an array of micropillars for filtering out large particles from the samples. Additionally, the porous structure of the yarn provides a secondary screening mechanism. The resulting system is compact, and reliable, and was successfully applied to the detection of Escherichia coli (E. coli) in two different types of berries using quantitative polymerase chain reaction (qPCR). The platform demonstrated a detection limit of 5 CFU g
-1 , showcasing its effectiveness in rapid and sensitive pathogen detection., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)- Published
- 2023
- Full Text
- View/download PDF
32. 3D printed integrated nanoporous membranes for electroextraction of DNA.
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Kalathil Balakrishnan H, Lee SM, Dumée LF, Doeven EH, Alexander R, Yuan D, and Guijt RM
- Subjects
- Printing, Three-Dimensional, Nucleic Acid Amplification Techniques, DNA, Escherichia coli, Nanopores
- Abstract
3D printing is established as an alternative microfabrication approach, and while printer resolution limits the direct 3D printing of pore features in the micron/submicron range, the use of nanoporous materials allows for the integration of porous membranes in 3D printed devices. Here, nanoporous membranes were formed by digital light projection (DLP) 3D printing using a polymerization-induced phase separation (PIPS) resin formulation. A functionally integrated device was fabricated using resin exchange following a simple, semi-automated manufacturing process. Printing of porous materials from a PIPS resin formulations based on polyethylene glycol diacrylate 250 as monomer was investigated by varying exposure time, photoinitiator concentration, and porogen content to yield materials with average pore size varying from 30-800 nm. Aiming for printing a size-mobility trap for electrophoretic extraction of deoxyribonucleic acid (DNA), conditions for printing materials with a mean pore size of 346 nm and 30 nm were selected for integration in a fluidic device using a resin exchange approach. Under optimized conditions (12.5 V for 20 min), cell concentrations as low as 10
3 cells per mL were detected following amplification of the extract by quantitative polymerase chain reaction (qPCR) at a Cq of 29. The efficacy of the size/mobility trap formed by the two membranes is demonstrated by detecting DNA concentrations equivalent to the input detected in the extract while removing 73% of the protein in the lysate. The DNA extraction yield was not statistically different from that obtained using a spin column, but manual handling and equipment needs were significantly reduced. This study demonstrates that nanoporous membranes with tailored properties can be integrated into fluidic devices using a simple manufacturing process based on resin exchange DLP. The process was used to manufacture a size-mobility trap and applied for the electroextraction and purification of DNA from E. coli lysate with reduced processing time, manual handling, and equipment needs compared with a commercially sourced DNA extraction kit. Combining manufacturability and portability with ease of use, the approach has demonstrated potential for manufacturing and using devices used in point-of-need testing for diagnostic nucleic acid amplification testing.- Published
- 2023
- Full Text
- View/download PDF
33. Closed-loop control systems for pumps used in portable analytical systems.
- Author
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Naz SA, Huynh VT, Doeven EH, Adams S, Kouzani A, and Guijt RM
- Subjects
- Reproducibility of Results, Syringes, Algorithms
- Abstract
The demand for accurate control of the flowrate/pressure in chemical analytical systems has given rise to the adoption of mechatronic approaches in analytical instruments. A mechatronic device is a synergistic system which combines mechanical, electronic, computer and control components. In the development of portable analytical devices, considering the instrument as a mechatronic system can be useful to mitigate compromises made to decrease space, weight, or power consumption. Fluid handling is important for reliability, however, commonly utilized platforms such as syringe and peristaltic pumps are typically characterized by flow/pressure fluctuations and slow responses. Closed loop control systems have been used effectively to decrease the difference between desired and realized fluidic output. This review discusses the way control systems have been implemented for enhanced fluidic control, categorized by pump type. Advanced control strategies used to enhance the transient and the steady state responses are discussed, along with examples of their implementation in portable analytical systems. The review is concluded with the outlook that the challenge in adequately expressing the complexity and dynamics of the fluidic network as a mathematical model has yielded a trend towards the adoption of experimentally informed models and machine learning approaches., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)
- Published
- 2023
- Full Text
- View/download PDF
34. Recent developments in digital light processing 3D-printing techniques for microfluidic analytical devices.
- Author
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Amini A, Guijt RM, Themelis T, De Vos J, and Eeltink S
- Subjects
- Lab-On-A-Chip Devices, Drug Delivery Systems, Microfluidics, Printing, Three-Dimensional
- Abstract
Digital light processing (DLP) 3D printing is rapidly advancing and has emerged as a powerful additive manufacturing approach to fabricate analytical microdevices. DLP 3D-printing utilizes a digital micromirror device to direct the projected light and photopolymerize a liquid resin, in a layer-by-layer approach. Advances in vat and lift design, projector technology, and resin composition, allow accurate fabrication of microchannel structures as small as 18 × 20 µm. This review describes the latest advances in DLP 3D-printing technology with respect to instrument set-up and resin formulation and highlights key efforts to fabricate microdevices targeting emerging (bio-)analytical chemistry applications, including colorimetric assays, extraction, and separation., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)
- Published
- 2023
- Full Text
- View/download PDF
35. Novel Boundary Lubrication Mechanisms from Molecular Pillows of Lubricin Brush-Coated Graphene Oxide Nanosheets.
- Author
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Russo MJ, Han M, Menon NG, Quigley AF, Kapsa RMI, Moulton SE, Guijt RM, M Silva S, Schmidt TA, and Greene GW
- Subjects
- Animals, Friction, Graphite, Lubrication, Mammals, Glycoproteins chemistry, Lubricants
- Abstract
There are numerous biomedical applications where the interfacial shearing of surfaces can cause wear and friction, which can lead to a variety of medical complications such as inflammation, irritation, and even bacterial infection. We introduce a novel nanomaterial additive comprised of two-dimensional graphene oxide nanosheets (2D-NSCs) coated with lubricin (LUB) to reduce the amount of tribological stress in biomedical settings, particularly at low shear rates where boundary lubrication dominates. LUB is a glycoprotein found in the articular joints of mammals and has recently been discovered as an ocular surface boundary lubricant. The ability of LUB to self-assemble into a "telechelic" brush layer on a variety of surfaces was exploited here to coat the top and bottom surfaces of the ultrathin 2D-NSCs in solution, effectively creating a biopolymer-coated nanosheet. A reduction in friction of almost an order of magnitude was measured at a bioinspired interface. This reduction was maintained after repeated washing (5×), suggesting that the large aspect ratio of the 2D-NSCs facilitates effective lubrication even at diluted concentrations. Importantly, and unlike LUB-only treatment, the lubrication effect can be eliminated over 15 rinsing cycles, suggesting that the LUB-coated 2D-NSCs do not exhibit any binding interactions with the shearing surfaces. The effective lubricating properties of the 2D-NSCs combined with full reversibility through rinsing make the LUB-coated 2D-NSCs an intriguing candidate as a lubricant for biomedical applications.
- Published
- 2022
- Full Text
- View/download PDF
36. Inexpensive portable capillary electrophoresis instrument for Monitoring Zinc(II) in remote areas.
- Author
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Zhang M, Smejkal P, Bester N, Robertson JC, Atia MA, Townsend AT, Guijt RM, and Breadmore MC
- Subjects
- Australia, Metals, Electrophoresis, Capillary methods, Zinc
- Abstract
A compact, inexpensive capillary electrophoresis instrument was developed for monitoring metal ions and evaluated for Zn(II) in remote contaminated locations in western Tasmania, Australia. The portable instrument, measuring 21 cm x 10 cm x 7 cm, was powered from the USB port of a laptop computer and built from off-the-shelf components costing ∼$1200 USD. Electrophoretic separations were conducted using a fused silica capillary (10-50 µm I.D.), applying 8.5 kV over capillaries ranging from 25 cm to 40 cm in length. The capillary inlet was connected with an electrically grounded cross-piece as flow-through injection interface. Automated fluidic management was achieved by controlling four mini peristaltic pumps and a solenoid valve. Detection was realised using a purpose-built visible LED absorption detector, optimised for the detection of Co(II), Cu(II) and Zn(II) after complexation with 4-(2-Pyridylazo) resorcinol (PAR). Limits of detection of sub-µM were obtained. The instrument was tested for continuous operation in the laboratory for up to 3 months, and relative standard deviations of <5.4% were found over 945 consecutive injections. In the field, the system was able to measure 106 samples within 11 h, the time it can be powered from the laptop computer. As Field measurement of Zn(II) in western Tasmania was demonstrated to show capability for on-site metal testing., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interest., (Copyright © 2022. Published by Elsevier B.V.)
- Published
- 2022
- Full Text
- View/download PDF
37. An improved nucleic acid sequence-based amplification method mediated by T4 gene 32 protein.
- Author
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Nai YH, Doeven EH, and Guijt RM
- Subjects
- DNA, RNA, Sensitivity and Specificity, Nucleic Acid Amplification Techniques, Self-Sustained Sequence Replication methods
- Abstract
The uptake of Nucleic Acid Sequence-Based Amplification (NASBA) for point of care testing may be hindered by a complexity in the workflow due the requirement of a thermal denaturation step to initiate the cyclic isothermal amplification before the addition of the amplification enzymes. Despite reports of successful enhancement of other DNA and RNA amplification methods using DNA and RNA binding proteins, this has not been reported for NASBA. Here, three single-stranded binding proteins, RecA, Extreme Thermostable Single-stranded binding protein (ET SSB) and T4 gene gp32 protein (gp32), were incorporated in NASBA protocol and used for single pot, one-step NASBA at 41 °C. Indeed, all SSBs showed significantly improved amplifications compared with the 2-step process, but only gp32 showed no non-specific aberrant amplification, and slightly improved the time-to-positivity in comparison with the conventional NASBA. For synthetic HIV-1 RNA, gp32 was found to improve the time-to-positivity (ttp) by average of 13.6% of one-step NASBA and 6.7% of conventional NASBA for the detection of HIV-1 RNA, showing its potential for simplifying the workflow as desirable for point of care applications of NASBA., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2022
- Full Text
- View/download PDF
38. 3D printing for the integration of porous materials into miniaturised fluidic devices: A review.
- Author
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Balakrishnan HK, Doeven EH, Merenda A, Dumée LF, and Guijt RM
- Subjects
- Membranes, Polymers, Porosity, Lab-On-A-Chip Devices, Printing, Three-Dimensional
- Abstract
Porous materials facilitate the efficient separation of chemicals and particulate matter by providing selectivity through structural and surface properties and are attractive as sorbent owing to their large surface area. This broad applicability of porous materials makes the integration of porous materials and microfluidic devices important in the development of more efficient, advanced separation platforms. Additive manufacturing approaches are fundamentally different to traditional manufacturing methods, providing unique opportunities in the fabrication of fluidic devices. The complementary 3D printing (3DP) methods are each accompanied by unique opportunities and limitations in terms of minimum channel size, scalability, functional integration and automation. This review focuses on the developments in the fabrication of 3DP miniaturised fluidic devices with integrated porous materials, focusing polymer-based methods including fused filament fabrication (FFF), inkjet 3D printing and digital light projection (DLP). The 3DP methods are compared based on resolution, scope for multimaterial printing and scalability for manufacturing. As opportunities for printing pores are limited by resolution, the focus is on approaches to incorporate materials with sub-micron pores to be used as membrane, sorbent or stationary phase in separation science using Post-Print, Print-Pause-Print and In-Print processes. Technical aspects analysing the efficiency of the fabrication process towards scalable manufacturing are combined with application aspects evaluating the separation and/or extraction performance. The review is concluded with an overview on achievements and opportunities for manufacturable 3D printed membrane/sorbent integrated fluidic devices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
39. Sheathless Separation of Cyanobacterial Anabaena by Shape Using Viscoelastic Microfluidics.
- Author
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Yuan D, Yan S, Zhang J, Guijt RM, Zhao Q, and Li W
- Subjects
- Cell Separation, Ecosystem, Microfluidics, Anabaena, Cyanobacteria
- Abstract
Cyanobacteria have a wide range of impact on natural ecosystems, and have been recognized as potentially rich sources of pharmacological and structurally interesting secondary metabolites. To better understand the basic molecular processes and mechanisms that influence and regulate the growth (like length) of cyanobacteria, or connections between environment, genotype, and phenotype, it would be essential to separate shape-synchronized cyanobacterial cell populations with relatively uniform length and size. This work proposes a novel and efficient method to separate cyanobacterial Anabaena by shape (rod aspect ratio) using viscoelastic microfluidics in a straight channel with expansion-contraction cavity arrays (ECCA channel). The biocompatible viscoelastic solutions with dissolved polymer would induce a combined effect of inertial lift force, elastic force, and secondary drag force for Anabaena flowing in it. Therefore, Anabaena with different lengths reach different lateral equilibrium positions and flow out from different outlets. Factors including flow rate, fluid viscoelasticity, channel structure, and length on the shape-based cell separation were studied systematically. This work, for the first time, demonstrates continuous and sheathless shape-based separation of cyanobacteria using viscoelastic microfluidics. Moreover, its ability to manipulate objects with different morphologies and with a size of >100 μm will extend the capability of microfluidics to a completely new field that has never been reached and would be attractive across a range of new applications.
- Published
- 2021
- Full Text
- View/download PDF
40. Facile fabrication of micro-/nanostructured, superhydrophobic membranes with adjustable porosity by 3D printing.
- Author
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Mayoussi F, Doeven EH, Kick A, Goralczyk A, Thomann Y, Risch P, Guijt RM, Kotz F, Helmer D, and Rapp BE
- Abstract
Porous membranes with special wetting properties have attracted great interest due to their various functions and wide applications, including water filtration, selective oil/water separation and oil skimming. Special wetting properties such as superhydrophobicity can be achieved by controlling the surface chemistry as well as the surface topography of a substrate. Three-dimensional (3D) printing is a promising method for the fast and easy generation of various structures. The most common method for 3D printing of superhydrophobic materials is a two-step fabrication process: 3D printing of user-defined topographies, such as surface structures or bulk porosity, followed by a chemical post-processing with low-surface energy chemicals such as fluorinated silanes. Another common method is using a hydrophobic polymer ink to print intricate surface structures. However, the resolution of most common printers is not sufficient to produce nano-/microstructured textures, moreover, the resulting delicate surface micro- or nanostructures are very prone to abrasion. Herein, we report a simple approach for 3D printing of superhydrophobic micro-/nanoporous membranes in a single step, combining the required topography and chemistry. The bulk porosity of this material, which we term "Fluoropor", makes it insensitive to abrasion. To achieve this, a photocurable fluorinated resin is mixed with a porogen mixture and 3D printed using a stereolithography (SLA) printing process. This way, micro-/nanoporous membranes with superhydrophobic properties with static contact angles of 164° are fabricated. The pore size of the membranes can be adjusted from 30 nm to 300 nm by only changing the porogen ratio in the mixture. We show the applicability of the printed membranes for oil/water separation and the formation of Salvinia layers which are of great interest for drag reduction in maritime transportation and fouling prevention., Competing Interests: The authors declare no conflict of interest., (This journal is © The Royal Society of Chemistry.)
- Published
- 2021
- Full Text
- View/download PDF
41. Hyphenated sample preparation-electrospray and nano-electrospray ionization mass spectrometry for biofluid analysis.
- Author
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Mikhail IE, Tehranirokh M, Gooley AA, Guijt RM, and Breadmore MC
- Subjects
- Miniaturization, Solid Phase Microextraction methods, Specimen Handling, Spectrometry, Mass, Electrospray Ionization methods
- Abstract
Stand-alone electrospray ionization mass spectrometry (ESI-MS) has been advancing through enhancements in throughput, selectivity and sensitivity of mass spectrometers. Unlike traditional MS techniques which usually require extensive offline sample preparation and chromatographic separation, many sample preparation techniques are now directly coupled with stand-alone MS to enable outstanding throughput for bioanalysis. In this review, we summarize the different sample clean-up and/or analyte enrichment strategies that can be directly coupled with ESI-MS and nano-ESI-MS for the analysis of biological fluids. The overview covers the hyphenation of different sample preparation techniques including solid phase extraction (SPE), solid phase micro-extraction (SPME), slug flow micro-extraction/nano-extraction (SFME/SFNE), liquid extraction surface analysis (LESA), extraction electrospray, extraction using digital microfluidics (DMF), and electrokinetic extraction (EkE) with ESI-MS and nano-ESI-MS., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interests., (Copyright © 2021 Elsevier B.V. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
42. Antifouling Strategies for Electrochemical Biosensing: Mechanisms and Performance toward Point of Care Based Diagnostic Applications.
- Author
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Russo MJ, Han M, Desroches PE, Manasa CS, Dennaoui J, Quigley AF, Kapsa RMI, Moulton SE, Guijt RM, Greene GW, and Silva SM
- Subjects
- Electrodes, Point-of-Care Systems, Biosensing Techniques, Electrochemical Techniques
- Abstract
Although there exist numerous established laboratory-based technologies for sample diagnostics and analyte detection, many medical and forensic science applications require point of care based platforms for rapid on-the-spot sample analysis. Electrochemical biosensors provide a promising avenue for such applications due to the portability and functional simplicity of the technology. However, the ability to develop such platforms with the high sensitivity and selectivity required for analysis of low analyte concentrations in complex biological samples remains a paramount issue in the field of biosensing. Nonspecific adsorption, or fouling, at the electrode interface via the innumerable biomolecules present in these sample types (i.e., serum, urine, blood/plasma, and saliva) can drastically obstruct electrochemical performance, increasing background "noise" and diminishing both the electrochemical signal magnitude and specificity of the biosensor. Consequently, this review aims to discuss strategies and concepts used throughout the literature to prevent electrode surface fouling in biosensors and to communicate the nature of the antifouling mechanisms by which they operate. Evaluation of each antifouling strategy is focused primarily on the fabrication method, experimental technique, sample composition, and electrochemical performance of each technology highlighting the overall feasibility of the platform for point of care based diagnostic/detection applications.
- Published
- 2021
- Full Text
- View/download PDF
43. Scalable 3D printing method for the manufacture of single-material fluidic devices with integrated filter for point of collection colourimetric analysis.
- Author
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Keshan Balavandy S, Li F, Macdonald NP, Maya F, Townsend AT, Frederick K, Guijt RM, and Breadmore MC
- Abstract
Assembly and bonding are major obstacles in manufacturing of functionally integrated fluidic devices. Here we demonstrate a single-material 3D printed device with an integrated porous structure capable of filtering particulate matter for the colourimetric detection of iron from soil and natural waters. Selecting a PolyJet 3D printer for its throughput, integrated filters were created exploiting a phenomenon occurring at the interface between the commercially available build material (Veroclear-RGD810) and water-soluble support material (SUP707). The porous properties were tuneable by varying the orientation of the print head relative to the channel and by varying the width of the build material. Porous structures ranging from 100 to 200 μm in thickness separated the sample and reagent chambers, filtering particles larger than 15 μm in diameter. Maintaining the manufacturing throughput of the Polyjet printer, 221 devices could be printed in 1.5 h (∼25 s per device). Including the 12 h post-processing soak in sodium hydroxide to remove the solid support material, the total time to print and process 221 devices was 13.5 h (3.6 min per device), with a material cost of $2.50 each. The applicability of the fluidic device for point of collection analysis was evaluated using colourimetric determination of iron from soil slurry and environmental samples. Following the reduction of Fe
3+ to Fe2+ using hydroxylammonium chloride, samples were introduced to the fluidic device where particulate matter was retained by the filter, allowing for particulate-free imaging of the red complex formed with 1,10-phenanthroline using a smartphone camera. The calibration curve ranged from of 1-100 mg L-1 Fe2+ and good agreement (95%) was obtained between the point of collection device and Sector Field ICP-MS., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)- Published
- 2021
- Full Text
- View/download PDF
44. A Comparison of Commercially Available Screen-Printed Electrodes for Electrogenerated Chemiluminescence Applications.
- Author
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Kerr E, Alexander R, Francis PS, Guijt RM, Barbante GJ, and Doeven EH
- Abstract
We examined a series of commercially available screen-printed electrodes (SPEs) for their suitability for electrochemical and electrogenerated chemiluminescence (ECL) detection systems. Using cyclic voltammetry with both a homogeneous solution-based and a heterogeneous bead-based ECL assay format, the most intense ECL signals were observed from unmodified carbon-based SPEs. Three commercially available varieties were tested, with Zensor outperforming DropSens and Kanichi in terms of sensitivity. The incorporation of nanomaterials in the electrode did not significantly enhance the ECL intensity under the conditions used in this evaluation (such as gold nanoparticles 19%, carbon nanotubes 45%, carbon nanofibers 21%, graphene 48%, and ordered mesoporous carbon 21% compared to the ECL intensity of unmodified Zensor carbon electrode). Platinum and gold SPEs exhibited poor relative ECL intensities (16% and 10%) when compared to carbonaceous materials, due to their high rates of surface oxide formation and inefficient oxidation of tri- n -propylamine (TPrA). However, the ECL signal at platinum electrodes can be increased ∼3-fold with the addition of a surfactant, which enhanced TPrA oxidation due to increasing the hydrophobicity of the electrode surface. Our results also demonstrate that each SPE should only be used once, as we observed a significant change in ECL intensity over repeated CV scans and SPEs cannot be mechanically polished to refresh the electrode surface., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kerr, Alexander, Francis, Guijt, Barbante and Doeven.)
- Published
- 2021
- Full Text
- View/download PDF
45. 3D Printing: An Alternative Microfabrication Approach with Unprecedented Opportunities in Design.
- Author
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Balakrishnan HK, Badar F, Doeven EH, Novak JI, Merenda A, Dumée LF, Loy J, and Guijt RM
- Abstract
In the past decade, 3D printing technologies have been adopted for the fabrication of microfluidic devices. Extrusion-based approaches including fused filament fabrication (FFF), jetting technologies including inkjet 3D printing, and vat photopolymerization techniques including stereolithography (SLA) and digital light projection (DLP) are the 3D printing methods most frequently adopted by the microfluidic community. Each printing technique has merits toward the fabrication of microfluidic devices. Inkjet printing offers a good selection of materials and multimaterial printing, and the large build space provides manufacturing throughput, while FFF offers a great selection of materials and multimaterial printing but at lower throughput compared to inkjet 3D printing. Technical and material developments adopted from adjacent research fields and developed by the microfluidic community underpin the printing of sub-100 μm enclosed microchannels by DLP, but challenges remain in multimaterial printing throughput. With the feasibility of 3D printed microfluidics established, we look ahead at trends in 3D printing to gain insights toward the future of this technology beyond the sole prism of being an alternative fabrication approach. A shift in emphasis from using 3D printing for prototyping, to mimic conventionally manufactured outputs, toward integrated approaches from a design perspective is critically developed.
- Published
- 2021
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- View/download PDF
46. In-Syringe Electrokinetic Protein Removal from Biological Samples prior to Electrospray Ionization Mass Spectrometry.
- Author
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Mikhail IE, Tehranirokh M, Gooley AA, Guijt RM, and Breadmore MC
- Subjects
- Acetaminophen blood, Atenolol blood, Blood Proteins chemistry, Chlorpheniramine blood, Clomipramine blood, Humans, Kinetics, Naproxen blood, Pindolol blood, Spectrometry, Mass, Electrospray Ionization, Blood Proteins isolation & purification, Syringes
- Abstract
Here, an electrokinetic extraction (EkE) syringe is presented allowing for on-line electrokinetic removal of serum proteins before ESI-MS. The proposed concept is demonstrated by the determination of pharmaceuticals from human serum within minutes, with sample preparation limited to a 5× dilution of the sample in the background electrolyte (BGE) and application of voltage, both of which can be performed in-syringe. Signal enhancements of 3.6-32 fold relative to direct infusion of diluted serum and up to 10.8 fold enhancement, were obtained for basic and acidic pharmaceuticals, respectively. Linear correlations for the basic drugs by EkE-ESI-MS/MS were achieved, covering the necessary clinical range with LOQs of 5.3, 7.8, 6.1, and 17.8 ng mL
-1 for clomipramine, chlorphenamine, pindolol, and atenolol, respectively. For the acidic drugs, the EkE-ESI-MS LOQs were 3.1 μg mL-1 and 2.9 μg mL-1 for naproxen and paracetamol, respectively. The EkE-ESI-MS and EkE-ESI-MS/MS methods showed good accuracy (%found of 81 % to 120 %), precision (≤20 %), and linearity (r>0.997) for all the studied drugs in spiked serum samples., (© 2020 Wiley-VCH GmbH.)- Published
- 2020
- Full Text
- View/download PDF
47. The influence of electrolyte concentration on nanofractures fabricated in a 3D-printed microfluidic device by controlled dielectric breakdown.
- Author
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Islam MF, Yap YC, Li F, Guijt RM, and Breadmore MC
- Subjects
- Butadienes chemistry, Equipment Design, Proteins analysis, Proteins chemistry, Styrene chemistry, Electrolytes chemistry, Electrophoresis instrumentation, Microfluidic Analytical Techniques instrumentation, Microfluidic Analytical Techniques methods, Nanostructures chemistry, Printing, Three-Dimensional instrumentation
- Abstract
A three-dimensional-printed microfluidic device made of a thermoplastic material was used to study the creation of molecular filters by controlled dielectric breakdown. The device was made from acrylonitrile butadiene styrene by a fused deposition modeling three-dimensional printer and consisted of two V-shaped sample compartments separated by 750 µm of extruded plastic gap. Nanofractures were formed in the thin piece of acrylonitrile butadiene styrene by controlled dielectric breakdown by application voltage of 15-20 kV with the voltage terminated when reaching a defined current threshold. Variation of the size of the nanofractures was achieved by both variation of the current threshold and by variation of the ionic strength of the electrolyte used for breakdown. Electrophoretic transport of two proteins, R-phycoerythrin (RPE; <10 nm in size) and fluorescamine-labeled BSA (f-BSA; 2-4 nm), was used to monitor the size and transport properties of the nanofractures. Using 1 mM phosphate buffer, both RPE and f-BSA passed through the nanofractures when the current threshold was set to 25 µA. However, when the threshold was lowered to 10 µA or lower, RPE was restricted from moving through the nanofractures. When we increased the electrolyte concentration during breakdown from 1 to 10 mM phosphate buffer, BSA passed but RPE was blocked when the threshold was equal to, or lower than, 25 µA. This demonstrates that nanofracture size (pore area) is directly related to the breakdown current threshold but inversely related to the concentration of the electrolyte used for the breakdown process., (© 2020 Wiley-VCH GmbH.)
- Published
- 2020
- Full Text
- View/download PDF
48. Rapid Additive Manufacturing of 3D Geometric Structures via Dual-Wavelength Polymerization.
- Author
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Li F, Thickett SC, Maya F, Doeven EH, Guijt RM, and Breadmore MC
- Abstract
A dual-wavelength photopolymerization process is presented, allowing for the volumetric fabrication of complex geometries using a multistep process. The methacrylate-based resin contained 0.1 wt % camphorquinone/0.1 wt % ethyl 4-(dimethylamino) benzoate and 0.2 wt % bis[2-(ochlorophenyl)-4,5-diphenylimidazole] as photoinitiator (473 nm) and photoinhibitor (365 nm), respectively. The photoinitiator and photoinhibitor concentrations were optimized to allow for photocuring to full depth (4.6 mm) following an exposure time of 2 min solely by 473 nm light, but no curing occurred when 365 nm light was present due to photoinhibition. This resin was validated using one-step volumetric fabrication of two objects containing voids defined by the 365 nm irradiation region. Two more complex structures were printed in a step-by-step manner, relying on the dynamic control of the projection patterns of both 365 and 473 nm projectors, decreasing the print time from 20 min using a commercially available single wavelength resin printer to 2 min.
- Published
- 2020
- Full Text
- View/download PDF
49. One step multi-material 3D printing for the fabrication of a photometric detector flow cell.
- Author
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Cecil F, Guijt RM, Henderson AD, Macka M, and Breadmore MC
- Abstract
Optical detection is the most common detection mode for many analytical assays. Photometric detection systems and their integration with analytical systems usually require several assembly parts and manual alignment of the capillary/tubing which affects sensitivity and repeatability. 3D printing is an innovative technology for the fabrication of integrated complex detection systems. One step multi-material 3D printing has been explored to fabricate a photometric detector flow cell from optically transparent and opaque materials using a dual-head FDM 3D printer. Integration of the microchannel, the detection window and the slit in a single device eliminates the need for manual alignment of fluidic and optical components, and hence improves sensitivity and repeatability. 3D printing allowed for rapid design optimisation by varying the slit dimension and optical pathlength. The optimised design was evaluated by determining stray light, effective path length and the signal to noise ratio using orange G. The optimised flow cell with extended path length of 10 mm and 500 μm slit yielded 0.02% stray light, 89% effective path length and detection limit of 2 nM. The sensitivity was also improved by 80% in the process of optimisation, using a blue 470 nm LED as a light source., (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
50. Colour tuning and enhancement of gel-based electrochemiluminescence devices utilising Ru(ii) and Ir(iii) complexes.
- Author
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Soulsby LC, Doeven EH, Pham TT, Eyckens DJ, Henderson LC, Long BM, Guijt RM, and Francis PS
- Abstract
Combining luminophores in ratios that compensate for energy transfer provides a readily selectable range of new emission colours for gel-based electrochemiluminescence devices (ECLDs). A novel blue ECLD luminophore is also introduced and shown to enhance the intensity of the conventional green emitter through a mixed annihilation ECL mechanism. Peak-to-peak voltages were minimised using asymmetric potential pulse sequences, which increased the longevity of the ECLD emission.
- Published
- 2019
- Full Text
- View/download PDF
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