UV-Assisted Hyperbranched Poly(ß-amino ester) Modification of a Silica Membrane for Two-Step Microfluidic DNA Extraction from Blood.

Integrating nucleic acid extraction in amplification-based point-of-care diagnostics will be a significant feature for next-generation point-of-care virus detection devices. However, extracting DNA efficiently on a microfluidic chip poses many technological and commercialization challenges, including manual steps, multiple instruments, pretreatment processes, and the use of organic solvents (ethanol, IPA) that inhibit detection, which is not viable with routine testing such as viral load monitoring of transplant patients for post-operative care. This paper presents a microfluidic system capable of two-step DNA extraction from blood using a UV-assisted hyperbranched poly(ß-amino ester) (HPAE)-modified silica membrane for cytomegalovirus (CMV) detection in a rapid and instrument-free manner without the presence of amplification inhibitors. HPAEs of varying branch ratios were synthesized, screened, and coated on a silica membrane and bonded between two layers of poly(methyl methacrylate) (PMMA) substrates. Our system could selectively extract DNA from blood with an efficiency of 94% and a lower limit viral load of 300 IU/mL in 20 min. The extracted DNA was used as the template for real-time loop-mediated isothermal amplification (LAMP)-based detection of CMV and was found to produce a fluorescent signal intensity that was comparable with commercially extracted templates. This system can be integrated easily with a nucleic acid amplification system and used for routine rapid testing of viral load in patient blood samples.

Permanent Hydrophobic Surface Treatment Combined with Solvent Vapor-Assisted Thermal Bonding for Mass Production of Cyclic Olefin Copolymer Microfluidic Chips.

A hydrophobic surface modification followed by solvent vapor-assisted thermal bonding was developed for the fabrication of cyclic olefin copolymer (COC) microfluidic chips. The modifier species 1H,1H,2H,2H-perfluorooctyl trichlorosilane (FOTS) was used to achieve the entrapment functionalization on the COC surface, and a hydrophobic surface was developed through the formation of a Si-O-Si crosslink network. The COC surface coated with 40 vol % cyclohexane, 59 vol % acetone, and 1 vol % FOTS by ultrasonic spray 10 and 20 times maintained its hydrophobicity with the water contact angle increasing from ∼86 to ∼115° after storage for 3 weeks. The solvent vapor-assisted thermal bonding was optimized to achieve high bond strength and good channel integrity. The results revealed that the COC chips exposed to 60 vol % cyclohexane and 40 vol % acetone for 120 s have the highest bond strength, with a burst pressure of ∼17 bar, which is sufficient for microfluidics applications such as droplet generation. After bonding, the channel maintained its integrity without any channel collapse. The hydrophobicity was also maintained, proved by the water contact angle of ∼115° on the bonded film, as well as the curved shape of water flow in the chip channel by capillary test. The combined hydrophobic treatment and solvent bonding process show significant benefits for scale-up production compared to conventional hydrophilic treatment for bonding and hydrophobic treatment using surface grafting or chemical vapor deposition since it does not require nasty chemistry, long-term treatment, vacuum chamber, and can be integrated into production line easily. Such a process can also be extended to permanent hydrophilic treatment combined with the bonding process and will lay a foundation for low-cost mass production of plastic microfluidic cartridges.

Perspectives in translating microfluidic devices from laboratory prototyping into scale-up production.

Transforming lab research into a sustainable business is becoming a trend in the microfluidic field. However, there are various challenges during the translation process due to the gaps between academia and industry, especially from laboratory prototyping to industrial scale-up production, which is critical for potential commercialization. In this Perspective, based on our experience in collaboration with stakeholders, e.g., biologists, microfluidic engineers, diagnostic specialists, and manufacturers, we aim to share our understanding of the manufacturing process chain of microfluidic cartridge from concept development and laboratory prototyping to scale-up production, where the scale-up production of commercial microfluidic cartridges is highlighted. Four suggestions from the aspect of cartridge design for manufacturing, professional involvement, material selection, and standardization are provided in order to help scientists from the laboratory to bring their innovations into pre-clinical, clinical, and mass production and improve the manufacturability of laboratory prototypes toward commercialization.

A centrifugal microfluidic pressure regulator scheme for continuous concentration control in droplet-based microreactors.

Droplet microfluidics is an emerging tool in many biological and chemical application areas such as digital polymerase chain reaction (PCR) and in vitro diagnosis because of its extremely small sample volume and wide range of possibilities for on-demand adjustment of droplet properties. Although centrifugal microfluidics has been reported as a viable scheme for droplet generation, there is not much progress as far as droplet manipulation and droplet-based reactions are concerned. In this paper, we report a microfluidic pressure regulator scheme along with the use of microcapillaries for periodic droplet generation and the subsequent fusion. This scheme enables fine control over droplet generation and the fusion process by varying the rotational frequency. To control the solution concentration in droplets, we have implemented several fusion devices, including one-to-one mode using a symmetric structure and ratio-adjustable mode with an asymmetric structure. As an application example, we performed cell transfection using the reported droplet-based technique, which resulted in considerable improvement in terms of transfection efficiency compared to the traditional bulk approach. In another example, we synthesized quasi-2D perovskites with controllable compositions and tunable photoluminescence peaks, thus confirming the volumetric accuracy of this approach down to the nano-liter scale. Compared to the common pressure pulsation approach, our centrifugal force actuation scheme offers the advantages of compactness and highly parallel batch processing. We anticipate that the new scheme will find many applications in cell biology and chemical synthesis.

Target trapping and in situ single-cell genetic marker detection with a focused optical beam.

Optical trapping of single particles or cells with the capability of in situ bio-sensing or genetic profiling opens the possibility of rapid screening of biological specimens. However, common optical tweezers suffer from the lack of long-range forces. Consequently, their application areas are predominantly limited to target manipulation instead of biological diagnostics. To solve this problem, we herein report an all-in-one approach by combining optical forces and convective drag forces generated through localized optothermal effect for long-range target manipulation. The device consists of a 2D array of gold coated polydimethylsiloxane (PDMS) micro-wells, which are immersed by colloidal particles or cell solution. Upon excitation of a 785-nm laser, the hydrodynamic convective force and optical forces will drag the targets of interest into their designated micro-wells. Moreover, the plasmonic thermal dissipation provides a constant temperature environment for following cell analysis procedures of cell isolation, lysis and isothermal nucleic acid amplification for the detection of genetic markers. With the merits of fabrication simplicity, short sample-to-answer cycle time and the compatibility with optical microscopes, the reported technique offers an attractive and highly versatile approach for on-site single cell analysis systems.

Real-time multi-channel SPR sensing based on DMD-enabled angular interrogation.

This paper reports a digital micro-mirror device (DMD)-enabled real-time multi-channel biosensing system based on angular interrogation surface plasmon resonance (SPR). In the experiments, angular scanning is achieved by a DMD that facilitates SPR measurements using a single-point photodetector. In the four-channel measurement setup, real-time monitoring of bovine serum albumin (BSA) and anti-BSA binding interactions is performed at various concentration levels. The experimental results have verified that the system has a resolution of 3.54 × 10-6 RIU (refractive index unit); and a detection limit of 9 ng/mL. The new DMD-based SPR interrogation system presents a new design route for practical solid-state SPR biosensing with a user-selectable range of interrogation, enhanced signal-to-noise ratio, and fast data throughput.

Thermal gradient induced tweezers for the manipulation of particles and cells.

Optical tweezers are a well-established tool for manipulating small objects. However, their integration with microfluidic devices often requires an objective lens. More importantly, trapping of non-transparent or optically sensitive targets is particularly challenging for optical tweezers. Here, for the first time, we present a photon-free trapping technique based on electro-thermally induced forces. We demonstrate that thermal-gradient-induced thermophoresis and thermal convection can lead to trapping of polystyrene spheres and live cells. While the subject of thermophoresis, particularly in the micro- and nano-scale, still remains to be fully explored, our experimental results have provided a reasonable explanation for the trapping effect. The so-called thermal tweezers, which can be readily fabricated by femtosecond laser writing, operate with low input power density and are highly versatile in terms of device configuration, thus rendering high potential for integration with microfluidic devices as well as lab-on-a-chip systems.