CE Chip Research Articles

On-chip Paper Electrophoresis for Ultrafast Screening of Infectious Diseases

The outbreak of new viral strains promotes advances in universal diagnostic techniques for detecting infectious diseases with unknown viral sequence. Long double-stranded RNA (dsRNA), a hallmark of infections, serves as a virus marker for prompt detection of viruses with unknown genomes. Here, we report on-chip paper electrophoresis for ultrafast screening of infectious diseases. Negatively charged RNAs pass through the micro and nanoscale pores of cellulose in order of size under an external electric field applied to the paper microfluidic channel. Quantitative separation of long dsRNA mimicking poly I:C was analyzed from 1.67 to 33 ng·μL−1, which is close to the viral dsRNA concentration in infected cells. This paper-based capillary electrophoresis chip (paper CE chip) can provide a new diagnostic platform for ultrafast viral disease detection at the point-of-care (POC) level.

Integrated chip electrophoresis and magnetic particle isolation used for detection of hepatitis B virus oligonucleotides

Rapid and sensitive detection is a key step in the effective and early response to the global hazard of various viral diseases. In this study, an integrated isolation of hepatitis B virus (HBV)-specific DNA fragment by magnetic nanoparticles (MNPs) and its immediate analysis by microchip CGE was performed. Microfluidic CE chip was used to accommodate the complete process of viral DNA isolation by MNPs including hybridization and thermal denaturation followed by CE separation. Beforehand, calibration curves of HBV fragments were constructed. For isolation by MNPs, specific streptavidin-biotin interaction was used to bind complementary HBV fragment to magnetic particles. After analysis of isolated HBV by regular MNPs method, innovative approach was performed. The commercial CE chip (Bio-rad) was successfully used to execute HBV fragment isolation. Detection using LIF with detection limit of 1 ng/mL was accomplished.

Contactless conductivity detection of small ions in a surface micro‐machined CE chip

A microchip is presented which is capable of CE separations and is built using exclusively thin film deposition techniques, fully compatible with microelectronics batch processing. Standard photolithography provides control of the spacing between electrodes used in conductivity measurement and overall channel geometry. Fluid channels are arranged as a double-T injector with a 50 microm offset at the arm intersection. The chip's performance was tested using concentrations of sodium chloride and calcium chloride ranging from 1 microM to 1 mM in a 5-mM MES/histidine buffer. Separations were performed by applying different voltages to reservoirs positioned at the four fluid channel openings. Conductivity detection was performed by applying a small AC voltage (1 Vrms) to the insulated electrodes positioned inside the fluid channels. A computer running LabVIEW controlled the AC signal generation, data acquisition and storage. Measurements indicated that the chip's detection limit was below 1 microM for both sodium and calcium cations.

Integrated Microfluidic System for Rapid Forensic DNA Analysis: Sample Collection to DNA Profile

We demonstrate a conduit for the delivery of a step change in the DNA analysis process: A fully integrated instrument for the analysis of multiplex short tandem repeat DNA profiles from reference buccal samples is described and is suitable for the processing of such samples within a forensic environment such as a police custody suite or booking office. The instrument is loaded with a DNA processing cartridge which incorporates on-board pumps and valves which direct the delivery of sample and reagents to the various reaction chambers to allow DNA purification, amplification of the DNA by PCR, and collection of the amplified product for delivery to an integral CE chip. The fluorescently labeled product is separated using micro capillary electrophoresis with a resolution of 1.2 base pairs (bp) allowing laser induced fluorescence-based detection of the amplified short tandem repeat fragments and subsequent analysis of data to produce a DNA profile which is compatible with the data format of the UK DNA database. The entire process from taking the sample from a suspect, to database compatible DNA profile production can currently be achieved in less than 4 h. By integrating such an instrument and microfluidic cartridge with the forensic process, we believe it will be possible in the near future to process a DNA sample taken from an individual in police custody and compare the profile with the DNA profiles held on a DNA Database in as little as 3 h.

A microfabricated CE chip for DNA pre‐concentration and separation utilizing a normally closed valve

A simple, sequential DNA pre-concentration and separation method by using a micro-CE chip integrated with a normally closed valve, which is activated by pneumatic suction, has been developed. The CE chip is fabricated using PDMS. A surface treatment technique for coating a polymer bilayer with an anionic charge is applied to modify the surface of the microchannel. A normally closed valve with anionic surface charges forms a nanoscale channel that only allows the passage of electric current but traps the DNA samples so that they are pre-concentrated. After the pre-concentration step, a pneumatic suction force is applied to the normally closed valve. This presses down the valve membrane, which reconnects the channels. The DNA samples are then moved into a separation channel for further separation and analysis. Successful DNA pre-concentration and separation has been achieved. Fluorescent intensity at the pre-concentration area is increased by approximately 3570 times within 1.9 min of operation. The signals from the separation of phiX174 DNA/HaeIII markers are enhanced approximately 41 times after 100 s of pre-concentration time, as compared with the results using a traditional cross-shaped micro-CE chip. These results clearly demonstrate that successful DNA pre-concentration for signal enhancement and separation analysis can be performed by using this new micro-CE chip.

Physical analysis, trimming and editing of nanoscale IC function with backside FIB processing

Most well established IR-beam based failure analysis techniques and also conventional circuit edit procedures are facing severe challenges resulting from the aggressive downscaling of today’s IC technology. To allow for alternative strategies, novel CE and functional chip analysis techniques have been developed, all being based on backside FIB processing. Additionally, in depth characterization of FIB induced device alterations has shown that a >20% speed gain can be achieved with the proposed FIB thinning procedure. In contrast to all known techniques, this offers trimming of chip internal timing conditions on fully functional samples without being bound to pre-planned fuses or varactors. Based on various experimental results and physical device simulations, this paper briefly reviews the necessary FIB process for which the main focus lies on the FIB induced device alteration. Finally, the novel CE and analysis techniques will be discussed regarding their fields of application, benefits compared to established techniques and theoretical limitations.

Electromagnetic induction detector with a vertical coil for capillary electrophoresis and microfluidic chip

We report a new electromagnetic induction detector with two vertical inductors for capillary electrophoresis and microfluidic chip capillary electrophoresis. Compared with currently available detection methods, this new detector has several advantages, including simple construction, no complicated elements, ease of assembly and operation, and potential for universal applications. The conditions affecting the response of the detector, including the number of coil turns, the frequency and the effective voltage, were optimized. Its feasibility and performance were demonstrated by analyzing inorganic ions. The new detector showed a well-defined correlation between the ion concentrations and the responses (r=0.993–0.998), with a detection limit of 5.6μmolL−1 for Mn2+, as well as good reproducibility and stability for both CE and microfluidic chip CE.

Fabrication of Si-PDMS Low Voltage Capillary Electrophoresis Chip

This paper discusses the fabrication of Si-PDMS low voltage capillary electrophoresis chip (CE chip). Arrayed-electrode which is used to apply low separation voltage is fabricated along the sidewalls of the separation channel on the silicon based bottom part. Isolation trenches, which are placed surrounding the arrayed-electrode, insure the insulation between the arrayed-electrode, as well as arrayed-electrode and liquid in the micro channel. Polydimethylsilicone (PDMS) is used as the cover. PDMS and silicon based bottom part are reversible sealed to attain Si-PDMS low voltage CE chip. Experiments have been done to obtain optimum electrophoresis separation condition: separation voltage is 45V, switch time is 2s and the Phe and Lys electrophoresis separation is successful.

Cover Picture: Electrophoresis 6/2008

AbstractRegular issues provide a wide range of research and review articles covering all aspects of electrophoresis. Here you will find cutting‐edge articles on methods and theory, instrumentation, nucleic acids, CE and CEC, miniaturization and microfluidics, proteomics and two‐dimensional electrophoresis.In addition, issue no. 6 features a series of 7 important papers on "Microfluidics and Miniaturization" dealing with LCD‐based optoelectronic tweezers, cell sorting, single cell clone analysis and cultivation, integrated ITP stacking and gel electrophoresis, floating injection, electrokinetic flows on thermosensitive surfaces and flow velocity measurement in CE chip instruments.

Identification of respiratory pathogen Bordetella Pertussis using integrated microfluidic chip technology

Integrated PCR–CE chip technology has immense potential to be applied in clinical diagnostics. In this work we demonstrate the application of our integrated PCR–CE chip for the detection of the respiratory pathogen Bordetella pertussis. A series of experiments with varying cell concentrations (200,000–2 cfu) were performed to obtain the analytical detection limits of the chip. We find that the chip technology is well suited for sensitive detection of Bordetella pertussis, using genetic material from less than even 2 cfu. We also utilized an off-chip real-time PCR method to compare and validate our on-chip approach.

A sheathless poly(methyl methacrylate) chip‐CE/MS interface fabricated using a wire‐assisted epoxy‐fixing method

Using a wire-assisted epoxy-fixing method, a sheathless CE/MS interface on a poly-(methyl methacrylate) (PMMA) CE chip has been developed. The sheathless chip-CE/MS interface utilized a tapered fused-silica tip and the electrical connection was achieved through a layered coating of conductive rubber. The wire-assisted method provided facile alignment of channels between the PMMA CE chip and an external capillary sprayer without the need for micromachining. Because the wire was in the channel during fixing, the risk of channel blockage by the epoxy was avoided. This chip CE device has minimal dead volume because the interstitial spaces were filled by a fast-fixing epoxy resin. The performance of the chip-CE-ESI-MS device was demonstrated with the analysis of peptide mixtures.

CE chips fabricated by injection molding and polyethylene/thermoplastic elastomer film packaging methods

This study presents a new packaging method using a polyethylene/thermoplastic elastomer (PE/TPE) film to seal an injection-molded CE chip made of either poly(methyl methacrylate) (PMMA) or polycarbonate (PC) materials. The packaging is performed at atmospheric pressure and at room temperature, which is a fast, easy, and reliable bonding method to form a sealed CE chip for chemical analysis and biomedical applications. The fabrication of PMMA and PC microfluidic channels is accomplished by using an injection-molding process, which could be mass-produced for commercial applications. In addition to microfluidic CE channels, 3-D reservoirs for storing biosamples, and CE buffers are also formed during this injection-molding process. With this approach, a commercial CE chip can be of low cost and disposable. Finally, the functionality of the mass-produced CE chip is demonstrated through its successful separation of phiX174 DNA/HaeIII markers. Experimental data show that the S/N for the CE chips using the PE/TPE film has a value of 5.34, when utilizing DNA markers with a concentration of 2 ng/microL and a CE buffer of 2% hydroxypropyl-methylcellulose (HPMC) in Tris-borate-EDTA (TBE) with 1% YO-PRO-1 fluorescent dye. Thus, the detection limit of the developed chips is improved. Lastly, the developed CE chips are used for the separation and detection of PCR products. A mixture of an amplified antibiotic gene for Streptococcus pneumoniae and phiX174 DNA/HaeIII markers was successfully separated and detected by using the proposed CE chips. Experimental data show that these DNA samples were separated within 2 min. The study proposed a promising method for the development of mass-produced CE chips.

Quantitative analysis of thiols in consumer products on a microfluidic CE chip with fluorescence detection

A microchip CE-based method for the quantification of the thiols mercaptoethanoic acid (MAA) and 2-mercaptopropionic acid (2-MPA) in depilatory cream and cold wave lotions was developed. The thiols were first derivatized with the fluorogenic reagent ammonium-7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD-F). The derivatives were separated within only 20 s by microchip CE and detected by their fluorescence. Conventional CE with diode array detection and LC with fluorescence detection were used for validation. The internal standard 3-mercaptopropionic acid (3-MPA) provided RSDs of multiple injections of only 4% or less for the MCE approach. LOD is 2 microM, LOQ 6 microM, and the linear range comprises nearly three decades of concentration starting at the LOQ.

Rapid prototyping of microfluidic devices with a wax printer

We demonstrate a rapid and inexpensive approach for the fabrication of high resolution poly(dimethylsiloxane) (PDMS)-based microfluidic devices. The complete process of fabrication could be performed in several hours (or less) without any specialized equipment other than a consumer-grade wax printer. The channels produced by this method are of high enough quality that we are able to demonstrate the sizing and separation of DNA fragments using capillary electrophoresis (CE) with no apparent loss of resolution over that found with glass chips fabricated by conventional photolithographic methods. We believe that this method will greatly improve the accessibility of rapid prototyping methods.

Fabrication of PMMA micro CE chip using IPA assisted low-temperature bonding

Fabrication of PMMA micro CE chip using IPA assisted low-temperature bonding

Numerical analysis and experimental estimation of a low-leakage injection technique for capillary electrophoresis.

This paper presents an experimental and numerical investigation into the use of low-leakage injection techniques to deliver sample plugs within electrophoresis microchips. The study addresses the principal material transport mechanisms such as electrokinetic migration, fluid flow, and diffusion and gives detail analyses to the double-L injection technique, which employs electrokinetic manipulations to avoid sample leakage within the microchip. Electrical potential contour, velocity vector, and streamline distribution in the micro CE chip are successfully developed. Experimental and numerical testing results show the double-L injection technique is capable of reducing sample leakage within cross-form microfluidic chips. The current study confirms the double-L injection technique has an exciting potential for use in high-quality, high-throughput chemical analysis applications and in many other applications throughout the field of micro total analysis systems.