Electromigration methods including CE and ITP are attractive for incorporation in microfluidic devices because they are relatively easily adaptable to miniaturization. After its popularity in the 1970s, ITP has made a comeback in microfluidic format (μ‐ITP, micro‐ITP) driven by the advantages of the steady‐state boundary, the self‐focusing effect, and the ability to aid in preconcentrating analytes in the sample while removing matrix components. In this review, we provide an overview of the developments in the area of μ‐ITP in a context of the historic developments with a focus on recent developments in experimental and computational ITP and discuss possible future trends. The chip‑ITP areas and topics discussed in this review and the corresponding sections include: PC simulations and modeling, analytical μ‐ITP, preconcentration ITP, transient ITP, peak mode ITP, gradient elution ITP, and free‐flow ITP, while the conclusions provide a critical summary and outlook. The review also contains experimental conditions for μ‐ITP applications to real‐world samples from over 50 original journal publications.
Three-dimensional (3D) printing has emerged as a potential revolutionary technology for the fabrication of microfluidic devices. A direct experimental comparison of the three 3D printing technologies dominating microfluidics was conducted using a Y-junction microfluidic device, the design of which was optimized for each printer: fused deposition molding (FDM), Polyjet, and digital light processing stereolithography (DLP-SLA). Printer performance was evaluated in terms of feature size, accuracy, and suitability for mass manufacturing; laminar flow was studied to assess their suitability for microfluidics. FDM was suitable for microfabrication with minimum features of 321 ± 5 μm, and rough surfaces of 10.97 μm. Microfluidic devices >500 μm, rapid mixing (71% ± 12% after 5 mm, 100 μL/min) was observed, indicating a strength in fabricating micromixers. Polyjet fabricated channels with a minimum size of 205 ± 13 μm, and a surface roughness of 0.99 μm. Compared with FDM, mixing decreased (27% ± 10%), but Polyjet printing is more suited for microfluidic applications where flow splitting is not required, such as cell culture or droplet generators. DLP-SLA fabricated a minimum channel size of 154 ± 10 μm, and 94 ± 7 μm for positive structures such as soft lithography templates, with a roughness of 0.35 μm. These results, in addition to low mixing (8% ± 1%), showed suitability for microfabrication, and microfluidic applications requiring precise control of flow. Through further discussion of the capabilities (and limitations) of these printers, we intend to provide guidance toward the selection of the 3D printing technology most suitable for specific microfluidic applications.
In capillary electrophoresis (CE), the inner surface of fused-silica capillaries is commonly covalently modified with liquid silanes to control electroosmotic flow (EOF). This liquid phase deposition (LPD) approach is challenging for long and narrow-diameter capillaries (≥1 m, ≤25 μm ID) inhibiting commercial production. Here, we use chemical vapour deposition (CVD) to covalently modify capillaries with different silanes. Using a home-built CVD device, capillaries were modified with neutral (3-glycidyloxypropyl) trimethoxysilane (GPTMS), the weak base (3-aminopropyl) trimethoxysilane (APTMS), the weak acid 3-mercaptopropyltrimethoxysilane (MPTMS) and the neutral hydrophobic trichloro(1H,1H,2H,2H-perfluorooctyl) silane (PFOCTS). Gas-phase modification of GPTMS with acid and ammonia allowed further modification of the surface prior to molecular layer deposition (MLD) of poly(p-phenylene terephthalamide) (PPTA) using the self-limiting sequential reaction between terephthalaldehyde (TA) and p-phenylenediamine (PD) vapours.Capillaries coated with GPTMS by CVD showed a greater reduction in EOF at all pH values than the conventional LPD. APTMS showed a reduction of the EOF at pH 9, with EOF reversal observed below pH 6. MPTMS provided a slightly lower EOF than an unmodified capillary at high pH, and a slightly higher EOF at lower pH. PFOCTS provided the most consistent EOF as a function of pH. The deposition of successive layers of PPTA resulted in increased surface coverage of the polymer and a greater reduction in EOF at pH higher than 5. The stability of a 10 μm ID GPTMS coated capillary was tested at pH 8.8 in a 200 mM CHES/Tris BGE for the separation of inorganic anions. Over 1.5 months of continuous operation (≈4130 runs), the reproducibility of the apparent mobilities for chloride, nitrite, nitrate and sulfate were 2.43%, 2.56%, 2.63% and 3.05%, respectively. The intra-day and inter-day column-to-column reproducibility and batch-to-batch reproducibility for all the coated capillaries ranged between 0.34% and 3.95%.The study demonstrates the superior performance of CVD coating for suppressing the EOF compared to LPD allowing the easy modification of long lengths of narrow capillary. The variation in silane, and the ability of MLD to modify and control the surface chemistry, provides a simple and facile method for surface modification. The stability of these coatings will allow long-term capillary electrophoresis monitoring of water chemistry, such as for monitoring fertiliser run-off in natural waters.
Within commercially available CE-on-a-chip platforms, the Agilent Bioanalyzer 2100 is well established for applications tailored methods for DNA, RNA and protein analysis. The instrument is equipped with a red diode laser - and optionally a blue LED - for fluorescence detection. The glass chips are polymer encased and contain12 sample reservoirs on one chip enabling high throughput analysis. This work targeted broadening the applicability of the Bioanalyzer to a wider range of analytes. The glass chips manufactured for DNA analysis were applied for generic CE based on a number of choices of injection and detection parameters. For non-aqueous CE (NACE), the compatibility of the existing chips with solvents such as dimethylsulfoxide (DMSO) was tested and the suitability of this non-volatile, non-hazardous, non-aquous solvent for NACE was verified and demonstrated with NACE separations of basic fluorescent dyes. The applicability of the instrument is enhanced both for direct and for indirect fluorescence detection by the presence of two excitation light sources: red laser (635 nm) and blue LED (470 nm). Several highly fluorescent dyes including Nile Blue (NB) and fluorescein isothiocyanate (FITC) that can be used as labelling tags were used to determine the sensitivity and LOD values. The suitability of the chip-based CE for indirect LIF detection using fluorescent dye probes and direct LIF determination of NB-tagged carboxylic acids, FITC-tagged amino acids as well as derivatised carbohydrates, and other biomolecules was investigated and will be illustrated by examples.
Abstract Fluorescently labeled carbohydrates released from glycoproteins were separated using a commercially available microfluidic chip electrophoresis system. While the instrumentation was primarily designed for DNA analysis it was found that the application base can be easily expanded using the development software provided by the manufacturer. The carbohydrates were released by enzymatic digestion (PNGase F) from glycoproteins present in human plasma after boronic acid – lectin affinity enrichment. After fluorescent labeling with 8‐aminopyrene‐1,3,6‐trisulfonic acid the carbohydrates were separated based on capillary gel electrophoresis mechanism and detected by a fluorescence detector using a blue (470 nm) LED. The separation was completed in 40 s in a microfluidic channel of 14 mm length. Glucose ladder carbohydrate oligomers differing by one glucose unit were baseline separated up to a 20‐mer with the main limitation being the detection sensitivity. As expected, the observed resolution in these experiments did not approach that of standard CE with 20 times longer separation distance; however, the chip‐based analysis excelled in the speed of the separation. Similar electrophoretic profiles of glycans released from plasma glycoproteins were obtained using a standard CE equipment with 35 cm separation length and microfluidic chips with a separation distance of only 14 mm.
'/%3e%3c/defs%3e%3cpath fill='%23012169' d='M0 0h10080v5040H0z'/%3e%3cpath stroke='%23fff' stroke-width='.6' d='m0 0 6 3m0-3L0 3' clip-path='url(%23b)' transform='scale(840)'/%3e%3cpath stroke='%23e4002b' stroke-width='.4' d='m0 0 6 3m0-3L0 3' clip-path='url(%23c)' transform='scale(840)'/%3e%3cpath stroke='%23fff' stroke-width='840' d='M2520 0v2520M0 1260h5040'/%3e%3cpath stroke='%23e4002b' stroke-width='504' d='M2520 0v2520M0 1260h5040'/%3e%3cg fill='%23fff'%3e%3cuse xlink:href='%23d' x='2520' y='3780'/%3e%3cuse xlink:href='%23a' x='7560' y='4200'/%3e%3cuse xlink:href='%23a' x='6300' y='2205'/%3e%3cuse xlink:href='%23a' x='7560' y='840'/%3e%3cuse xlink:href='%23a' x='8680' y='1869'/%3e%3cuse xlink:href='%23e' x='8064' y='2730'/%3e%3c/g%3e%3c/svg%3e)
'/%3e%3cpath d='M200 752.362h200v300H200z' style='fill:%23fff;fill-opacity:1;stroke:none' transform='translate(0 -752.362)'/%3e%3cpath d='M400 752.362h200v300H400z' style='fill:%23ff883e;fill-opacity:1;stroke:none' transform='translate(0 -752.362)'/%3e%3c/svg%3e)