Kangning Ren

Materials for microfluidic chip fabrication

Through manipulating fluids using microfabricated channel and chamber structures, microfluidics is a powerful tool to realize high sensitive, high speed, high throughput, and low cost analysis. In addition, the method can establish a well-controlled microenivroment for manipulating fluids and particles. It also has rapid growing implementations in both sophisticated chemical/biological analysis and low-cost point-of-care assays. Some unique phenomena emerge at the micrometer scale. For example, reactions are completed in a shorter amount of time as the travel distances of mass and heat are relatively small; the flows are usually laminar; and the capillary effect becomes dominant owing to large surface-to-volume ratios. In the meantime, the surface properties of the device material are greatly amplified, which can lead to either unique functions or problems that we would not encounter at the macroscale. Also, each material inherently corresponds with specific microfabrication strategies and certain native properties of the device. Therefore, the material for making the device plays a dominating role in microfluidic technologies. In this Account, we address the evolution of materials used for fabricating microfluidic chips, and discuss the application-oriented pros and cons of different materials. This Account generally follows the order of the materials introduced to microfluidics. Glass and silicon, the first generation microfluidic device materials, are perfect for capillary electrophoresis and solvent-involved applications but expensive for microfabriaction. Elastomers enable low-cost rapid prototyping and high density integration of valves on chip, allowing complicated and parallel fluid manipulation and in-channel cell culture. Plastics, as competitive alternatives to elastomers, are also rapid and inexpensive to microfabricate. Their broad variety provides flexible choices for different needs. For example, some thermosets support in-situ fabrication of arbitrary 3D structures, while some perfluoropolymers are extremely inert and antifouling. Chemists can use hydrogels as highly permeable structural material, which allows diffusion of molecules without bulk fluid flows. They are used to support 3D cell culture, to form diffusion gradient, and to serve as actuators. Researchers have recently introduced paper-based devices, which are extremely low-cost to prepare and easy to use, thereby promising in commercial point-of-care assays. In general, the evolution of chip materials reflects the two major trends of microfluidic technology: powerful microscale research platforms and low-cost portable analyses. For laboratory research, chemists choosing materials generally need to compromise the ease in prototyping and the performance of the device. However, in commercialization, the major concerns are the cost of production and the ease and reliability in use. There may be new growth in the combination of surface engineering, functional materials, and microfluidics, which is possibly accomplished by the utilization of composite materials or hybrids for advanced device functions. Also, significant expanding of commercial applications can be predicted.

Whole-Teflon microfluidic chips

Although microfluidics has shown exciting potential, its broad applications are significantly limited by drawbacks of the materials used to make them. In this work, we present a convenient strategy for fabricating whole-Teflon microfluidic chips with integrated valves that show outstanding inertness to various chemicals and extreme resistance against all solvents. Compared with other microfluidic materials [e.g., poly(dimethylsiloxane) (PDMS)] the whole-Teflon chip has a few more advantages, such as no absorption of small molecules, little adsorption of biomolecules onto channel walls, and no leaching of residue molecules from the material bulk into the solution in the channel. Various biological cells have been cultured in the whole-Teflon channel. Adherent cells can attach to the channel bottom, spread, and proliferate well in the channels (with similar proliferation rate to the cells in PDMS channels with the same dimensions). The moderately good gas permeability of the Teflon materials makes it suitable to culture cells inside the microchannels for a long time.

Chemical Recognition in Cell-Imprinted Polymers

A glass slide covered with bacteria is pressed into another glass slide coated with partially cured polydimethylsiloxane (PDMS). The PDMS is hardened and the cells are removed to create a textured surface whose indentations preferentially capture the same type of bacteria when a mixture of bacteria is flowed over it. Overcoating the cell-imprinted PDMS with methylsilane groups causes the resulting surface to lose much of its ability to preferentially capture the imprinted bacteria, although the shapes of the imprints, measured by atomic force field microscopy, are shown to be hardly affected. We interpret this behavior as strong evidence that chemical recognition plays a dominant role in cell sorting with cell-imprinted PDMS polymer films.

Convenient Method for Modifying Poly(dimethylsiloxane) with Poly(ethylene glycol) in Microfluidics

This report describes a convenient and reproducible method for the covalent modification of poly(dimethylsiloxane) (PDMS) with poly(ethylene glycol) (PEG) chains to suppress nonspecific protein adsorption. PEG additives terminated with a vinyl group are added into the PDMS prepolymer; when the PDMS is thermally cured, the vinyl group reacts with the silane groups on the PDMS, which covalently link the PEG group to the PDMS network. The PEG-modified PDMS surfaces are characterized with FT-IR, X-ray photoelectron spectroscopy (XPS), and contact angle measurement. We also examined the modified PDMS for on-chip capillary electrophoresis and its capability of resisting nonspecific protein adsorption using bovine serum albumin (BSA) as a model. Based on our study, a molecular mechanism is given to successfully explain the surface properties of the modified PDMS surfaces.

New materials for microfluidics in biology

With its continuous progress, microfluidics has become a key enabling technology in biological research. During the past few years, the major growth of microfluidics shifted to the introduction of new materials in making microfluidic chips, primarily driven by the demand of versatile strategies to interface microfluidics with biological cell studies. Although polydimethylsiloxane is still used as primary frame material, hydrogels have been increasingly employed in cell-culture related applications. Moreover, plastics and paper are attracting more attention in commercial device fabrication. Aiming to reflect this trend, current review focuses on the progress of microfluidic chip materials over the time span of January 2011 through June 2013, and provides critical discussion of the resulting major new tools in biological research.

Recent Developments in Microfluidics for Cell Studies

As a technique for precisely manipulating fluid at the micrometer scale, the field of microfluidics has experienced an explosive growth over the past two decades, particularly owing to the advances in device design and fabrication. With the inherent advantages associated with its scale of operation, and its flexibility in being incorporated with other microscale techniques for manipulation and detection, microfluidics has become a major enabling technology, which has introduced new paradigms in various fields involving biological cells. A microfluidic device is able to realize functions that are not easily imaginable in conventional biological analysis, such as highly parallel, sophisticated high-throughput analysis, single-cell analysis in a well-defined manner, and tissue engineering with the capability of manipulation at the single-cell level. Major advancements in microfluidic device fabrication and the growing trend of implementing microfluidics in cell studies are presented, with a focus on biological research and clinical diagnostics.

Fabrication of a microfluidic Ag/AgCl reference electrode and its application for portable and disposable electrochemical microchips

This report describes a convenient method for the fabrication of a miniaturized, reliable Ag/AgCl reference electrode with nanofluidic channels acting as a salt bridge that can be easily integrated into microfluidic chips. The Ag/AgCl reference electrode shows high stability with millivolt variations. We demonstrated the application of this reference electrode in a portable microfluidic chip that is connected to a USB‐port microelectrochemical station and to a computer for data collection and analysis. The low fabrication cost of the chip with the potential for mass production makes it disposable and an excellent candidate for real‐world analysis and measurement. We used the chip to quantitatively analyze the concentrations of heavy metal ions (Cd2+ and Pb2+) in sea water. We believe that the Ag/AgCl reference microelectrode and the portable electrochemical system will be of interest to people in microfluidics, environmental science, clinical diagnostics, and food research.

LprG-mediated surface expression of lipoarabinomannan is essential for virulence of Mycobacterium tuberculosis.

Mycobacterium tuberculosis employs various virulence strategies to subvert host immune responses in order to persist and cause disease. Interaction of M. tuberculosis with mannose receptor on macrophages via surface-exposed lipoarabinomannan (LAM) is believed to be critical for cell entry, inhibition of phagosome-lysosome fusion, and intracellular survival, but in vivo evidence is lacking. LprG, a cell envelope lipoprotein that is essential for virulence of M. tuberculosis, has been shown to bind to the acyl groups of lipoglycans but the role of LprG in LAM biosynthesis and localization remains unknown. Using an M. tuberculosis lprG mutant, we show that LprG is essential for normal surface expression of LAM and virulence of M. tuberculosis attributed to LAM. The lprG mutant had a normal quantity of LAM in the cell envelope, but its surface was altered and showed reduced expression of surface-exposed LAM. Functionally, the lprG mutant was defective for macrophage entry and inhibition of phagosome-lysosome fusion, was attenuated in macrophages, and was killed in the mouse lung with the onset of adaptive immunity. This study identifies the role of LprG in surface-exposed LAM expression and provides in vivo evidence for the essential role surface LAM plays in M. tuberculosis virulence. Findings have translational implications for therapy and vaccine development.

Convenient Method for Modifying Poly(dimethylsiloxane) To Be Airtight and Resistive against Absorption of Small Molecules

In this paper we present a simple and rapid method of modifying poly(dimethylsiloxane) (PDMS) surfaces with paraffin wax. PDMS that contains a layer of paraffin wax at its surface resists the absorption of hydrophobic molecules; we used fluorescence microscopy to confirm that paraffin-modified PDMS resists the absorption of rhodamine B. Furthermore, we demonstrated that microfluidic devices made from PDMS that contains a surface layer of paraffin wax prevent efficiently the transport of gas molecules through the bulk and into microchannels. We characterized the surface of PDMS that contains paraffin wax using the water contact angle, optical transmission, and X-ray photoelectron spectroscopy. We show that PDMS that contains paraffin wax can be substituted for native PDMS; specifically, we fabricated peristaltic valves in PDMS that contains paraffin wax, and the valves showed no degradation in performance after multiple open/close cycles. Finally, we show how to use PDMS that has been treated with paraffin wax as a mold for the fabrication of PDMS replicas; this approach avoids silanization of PDMS, which is a time-consuming step in soft lithography. The wax-modified PDMS channels also show performance superiro to that of bare PDMS in micellar electrokinetic chromatography for quantitative analysis.

Sorting Inactivated Cells Using Cell-Imprinted Polymer Thin Films

Previous work showed that cell imprinting in a poly(dimethylsiloxane) film produced artificial receptors to cells by template-assisted rearrangement of functional groups on the surface of the polymer thin film which facilitated cell capture in the polymer surface indentations by size, shape, and, most importantly, chemical recognition. We report here that inactivation of cells by treatment with formaldehyde (4%), glutaraldehyde (2%), or a combination of the two leads to markedly improved capture selectivity (a factor of 3) when cells to be analyzed are inactivated in the same manner. The enhanced capture efficiency compared to living cells results from two factors: (1) rigidification of the cell surface through cross-linking of amine groups by the aldehyde; and (2) elimination of chemicals excreted from living cells which interfere with the fidelity of the cell-imprinting process. Moreover, cell inactivation has the advantage of removing biohazard risks associated with working with virulent bacteria. These results are demonstrated using different strains of Mycobacterium tuberculosis.