Yuejun Kang

Paper-Based Microfluidic Electrochemical Immunodevice Integrated with Nanobioprobes onto Graphene Film for Ultrasensitive Multiplexed Detection of Cancer Biomarkers

To the best of our knowledge, this was the first report on the integration of a signal amplification strategy into a microfluidic paper-based electrochemical immunodevice for the multiplexed measurement of cancer biomarkers. Signal amplification was achieved through the use of graphene to modify the immunodevice surface to accelerate the electron transfer and the use of silica nanoparticles as a tracing tag to label the signal antibodies. Accurate, rapid, simple, and inexpensive point-of-care electrochemical immunoassays were demonstrated using a photoresist-patterned microfluidic paper-based analytical device (μPAD). Using the horseradish peroxidase (HRP)-O-phenylenediamine-H2O2 electrochemical detection system, the potential clinical applicability of this immunodevice was demonstrated through its ability to identify four candidate cancer biomarkers in serum samples from cancer patients. The novel signal-amplified strategy proposed in this report greatly enhanced the sensitivity of the detection of cancer biomarkers. In addition, the electrochemical immunodevice exhibited good stability, reproducibility, and accuracy and thus had potential applications in clinical diagnostics.

Dynamic aspects of electroosmotic flow in a cylindrical microcapillary

Abstract Dynamic aspects of the electroosmotic flow (EOF) in a cylindrical capillary are analysed. An analytical solution for electrostatic potential of the double layer has been derived by solving the complete Poisson–Boltzmann equation for arbitrary zeta-potentials under an analytical scheme. Transient EOF field in response to the application of time dependent electric field is obtained analytically by using Green’s function method. Specifically, sinusoidally alternating (AC) electric fields are used to study the effect of frequency-dependent oscillation on the EOF. Limiting cases of zero frequency and pulsed electric field are also discussed.

A paper-based microfluidic electrochemical immunodevice integrated with amplification-by-polymerization for the ultrasensitive multiplexed detection of cancer biomarkers

A novel signal amplification strategy for ultrasensitive multiplexed detection of cancer biomarkers using a paper-based microfluidic electrochemical immunodevice is described. Specifically, a controlled radical polymerization reaction is triggered after the capture of target molecules on the immunodevice surface. Growth of long chain polymeric materials provides numerous sites for subsequent horseradish peroxidase (HRP) coupling, which in turn significantly enhances electrochemical signal output. The signal was further amplified through the use of graphene to modify the immunodevice surface to accelerate the electron transfer. Activators generated electron transfer for atom transfer radical polymerization (AGET ATRP) was used in this study for its high efficiency in polymer grafting and better tolerance toward oxygen in air. Glycidyl methacrylate (GMA) was examined to provide excess epoxy groups for HRP coupling. In the electrochemical immunodevice, eight carbon working electrodes, as well as their conductive pads, were screen-printed on a piece of square paper, and the same Ag/AgCl reference and carbon counter electrodes were shared with another piece of square paper via stacking. Using the HRP-O-phenylenediamine-H2O2 electrochemical detection system, four cancer biomarkers: carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), cancer antigen 125 (CA125), and carbohydrate antigen 153 (CA153) were detected. A limit of detection of 0.01, 0.01, 0.05 and 0.05 ng mL(-1) was demonstrated, respectively. The results show that the proposed strategy offers great promises in providing a sensitive and cost-effective solution for biosensing applications.

Effects of dc-dielectrophoretic force on particle trajectories in microchannels

A method of controlling the particle trajectory in a microchannel is demonstrated. The method utilizes the dc-dielectrophoretic (dc-DEP) force created around an insulating hurdle in a microchannel under an applied dc electric field. This method does not require a complicated electrode array which is commonly used in the conventional ac-DEP system. The “proof-of-principle” experiments were carried out using a straight microchannel with a rectangle-shaped hurdle in the middle. The experiments showed that the trajectories of the micron-sized particles can be controlled by the DEP force under electric-field strengths of 5–20kV∕m. To compare with the experimental results, the particle motion was simulated using the Lagrangian tracking method, taking into consideration of the electrophoretic force, the dielectrophoretic force, and the dielectric interaction between the particle and the channel wall. The numerical simulation based on the finite-element method showed a reasonable agreement with the experimental data.

Surface Chemical Modification of Poly(dimethylsiloxane) for the Enhanced Adhesion and Proliferation of Mesenchymal Stem Cells

The surface chemistry of materials has an interactive influence on cell behavior. The optimal adhesion of mammalian cells is critical in determining the cell viability and proliferation on substrate surfaces. Because of the inherent high hydrophobicity of a poly(dimethylsiloxane) (PDMS) surface, cell culture on these surfaces is unfavorable, causing cells to eventually dislodge from the surface. Although physically adsorbed matrix proteins can promote initial cell adhesion, this effect is usually short-lived. Here, (3-aminopropyl)triethoxy silane (APTES) and cross-linker glutaraldehyde (GA) chemistry was employed to immobilize either fibronectin (FN) or collagen type 1 (C1) on PDMS. The efficiency of these surfaces to support the adhesion and viability of mesenchymal stem cells (MSCs) was analyzed. The hydrophobicity of the native PDMS decreased significantly with the mentioned surface functionalization. The adhesion of MSCs was mostly favorable on chemically modified PDMS surfaces with APTES + GA + protein. Additionally, the spreading area of MSCs was significantly higher on APTES + GA + C1 surfaces than on other unmodified/modified PDMS surfaces with C1 adsorption. However, there were no significant differences in the MSC spreading area on the unmodified/modified PDMS surfaces with FN adsorption. Furthermore, there was a significant increase in cell proliferation on the PDMS surface with APTES + GA + protein functionalization as compared to the PDMS surface with protein adsorption only. Therefore, the covalent surface chemical modification of PDMS with APTES + GA + protein could offer a more biocompatible platform for the enhanced adhesion and proliferation of MSCs. Similar strategies can be applied for other substrates and cell lines by appropriate combinations of self-assembly monolayers (SAMs) and extracellular matrix proteins.

Highly Specific and Ultrasensitive Graphene-Enhanced Electrochemical Detection of Low-Abundance Tumor Cells Using Silica Nanoparticles Coated with Antibody-Conjugated Quantum Dots

A dual signal amplification immunosensing strategy that offers high sensitivity and specificity for the detection of low-abundance tumor cells was designed. High sensitivity was achieved by using graphene to modify the immunosensor surface to accelerate electron transfer and quantum dot (QD)-coated silica nanoparticles as tracing tags. High specificity was further obtained by the simultaneous measurement of two disease-specific biomarkers on the cell surface using different QD-coated silica nanoparticle tracers. The immunosensor was constructed by covalently immobilized capture antibodies on a chitosan/electrochemically reduced graphene oxide film-modified glass carbon electrode. Cells were captured with a sandwich-type immunoreaction and the different QD-coated silica nanoparticle tracers were captured on the surface of the cells. Each biorecognition event yields a distinct voltammetric peak, which position and size reflects the corresponding identity and amount of the respective antigen. This strategy was vividly demonstrated by the simultaneous immunoassay of EpCAM and GPC3 antigens on the surface of the human liver cancer cell line Hep3B using anti-EpCAM-CdTe- and anti-GPC3-ZnSe-coated silica nanoparticle tracers. The two tracers gave comparable sensitivity, and the immunosensor exhibited high sensitivity and specificity with excellent stability, reproducibility, and accuracy, indicating its wide range of potential applications in clinical and molecular diagnostics.

Near-Infrared Squaraine Dye Encapsulated Micelles for in Vivo Fluorescence and Photoacoustic Bimodal Imaging

Combined near-infrared (NIR) fluorescence and photoacoustic imaging techniques present promising capabilities for noninvasive visualization of biological structures. Development of bimodal noninvasive optical imaging approaches by combining NIR fluorescence and photoacoustic tomography demands suitable NIR-active exogenous contrast agents. If the aggregation and photobleaching are prevented, squaraine dyes are ideal candidates for fluorescence and photoacoustic imaging. Herein, we report rational selection, preparation, and micelle encapsulation of an NIR-absorbing squaraine dye (D1) for in vivo fluorescence and photoacoustic bimodal imaging. D1 was encapsulated inside micelles constructed from a biocompatible nonionic surfactant (Pluoronic F-127) to obtain D1-encapsulated micelles (D1(micelle)) in aqueous conditions. The micelle encapsulation retains both the photophysical features and chemical stability of D1. D1(micelle) exhibits high photostability and low cytotoxicity in biological conditions. Unique properties of D1(micelle) in the NIR window of 800-900 nm enable the development of a squaraine-based exogenous contrast agent for fluorescence and photoacoustic bimodal imaging above 820 nm. In vivo imaging using D1(micelle), as demonstrated by fluorescence and photoacoustic tomography experiments in live mice, shows contrast-enhanced deep tissue imaging capability. The usage of D1(micelle) proven by preclinical experiments in rodents reveals its excellent applicability for NIR fluorescence and photoacoustic bimodal imaging.

Microfluidic differential resistive pulse sensors

This study demonstrates an on‐chip resistive pulse‐sensing scheme with a design of symmetric mirror channels, which significantly reduces the noise and achieves better signal‐to‐noise ratio. Polystyrene particles of different sizes have been detected with the developed sensing scheme and a record low volume ratio of the particle to the sensing channel, or 0.0004%, has been detected with particles of 520 nm in diameter in a sensing aperture of 50×16×20 μm3. This volume ratio is about ten times lower than the lowest volume ratio reported in the literature including that specified for commercial Coulter counters.

Continuous particle separation by size via AC-dielectrophoresis using a lab-on-a-chip device with 3-D electrodes

In this paper, we present a novel, simple lab‐on‐a‐chip device for continuous separation of particles by size. The device is composed of a straight rectangular microchannel connecting two inlet reservoirs and two exit reservoirs. Two asymmetric, 3‐D electrodes are embedded along the channel wall to generate a non‐uniform electrical field for dielectrophoresis. Particles with different sizes are collected at the different exit reservoirs. Main flow is induced by pressure difference between the inlet and the exit reservoirs. The device is used successfully for the separation of the 5 and 10 μm latex particles and for the separation of yeast cells and white blood cells. A numerical simulation based on Lagrangian tracking method is used to simulate the particle motion and the results showed a good agreement with the experimental data.

Simple surface engineering of polydimethylsiloxane with polydopamine for stabilized mesenchymal stem cell adhesion and multipotency.

Polydimethylsiloxane (PDMS) has been extensively exploited to study stem cell physiology in the field of mechanobiology and microfluidic chips due to their transparency, low cost and ease of fabrication. However, its intrinsic high hydrophobicity renders a surface incompatible for prolonged cell adhesion and proliferation. Plasma-treated or protein-coated PDMS shows some improvement but these strategies are often short-lived with either cell aggregates formation or cell sheet dissociation. Recently, chemical functionalization of PDMS surfaces has proved to be able to stabilize long-term culture but the chemicals and procedures involved are not user- and eco-friendly. Herein, we aim to tailor greener and biocompatible PDMS surfaces by developing a one-step bio-inspired polydopamine coating strategy to stabilize long-term bone marrow stromal cell culture on PDMS substrates. Characterization of the polydopamine-coated PDMS surfaces has revealed changes in surface wettability and presence of hydroxyl and secondary amines as compared to uncoated surfaces. These changes in PDMS surface profile contribute to the stability in BMSCs adhesion, proliferation and multipotency. This simple methodology can significantly enhance the biocompatibility of PDMS-based microfluidic devices for long-term cell analysis or mechanobiological studies.