Evaluation of Fluid Behaviors in a Pushbutton-Activated Microfluidic Device for User-Independent Flow Control.

Although numerous studies have been conducted to realize ideal point-of-care testing (POCT), the development of a user-friendly and user-independent power-free microfluidic platform is still a challenge. Among various methods, the finger-actuation method shows a promising technique that provides a user-friendly and equipment-free way of delivering fluid in a designated manner. However, the design criteria and elaborate evaluation of the fluid behavior of a pushbutton-activated microfluidic device (PAMD) remain a critical bottleneck to be widely adopted in various applications. In this study, we have evaluated the fluid behavior of the PAMD based on various parameters, such as pressing velocity and depth assisted by a press machine. We have further developed a user-friendly and portable pressing block that reduces user variation in fluid behavior based on the evaluation.

Characterization of PDMS Microchannels Using Horizontally or Vertically Formed 3D-Printed Molds by Digital Light Projection.

Three-dimensional (3D) printing is one of the promising technologies for the fabrication of microstructures due to its versatility, ease of fabrication, and low cost. However, the direct use of 3D-printed microstructure as a microchannel is still limited due to its surface property, biocompatibility, and transmittance. As an alternative, rapid prototyping of poly(dimethylsiloxane) (PDMS) from 3D-printed microstructures ensures both biocompatibility and efficient fabrication. We employed 3D-printed molds fabricated using horizontal and vertical arrangement methods with different slice thicknesses in a digital light projection (DLP)-based 3D printing process to replicate PDMS microchannels. The replicated PDMS structures were investigated to compare their optical transmittances and surface roughness. Interestingly, the optical transmittance of PDMS from the 3D-printed mold was significantly increased via bonding two single PDMS layers. To evaluate the applicability of the replicated PDMS devices from the 3D-printed mold, we performed droplet generation in the PDMS microchannels, comparing the same device from a conventional Si-wafer mold. This study provides a fundamental understanding of prototyping microstructures from the DLP-based 3D-printed mold.

On-chip microfluidic dual detection of amino acid metabolism disorders using cell-free protein synthesis.

Various metabolic diseases are associated with the accumulation of specific amino acids due to abnormal metabolic pathways, and thus can be diagnosed by measuring the level of amino acids in body fluids. However, present methods for amino acid analysis are not readily accessible because they require a complex experimental setup, expensive equipment, and a long processing time. Here, we present a dual sensing microfluidic device that enables fast, portable, and quantitative analysis of target amino acids, harnessing the biological mechanism of protein synthesis. In this device, the working principle of a finger-actuated pumping unit is applied, and the microchannels are designed to perform cell-free synthesis of a reporter protein in response to the target amino acids in the assay samples. Multiple steps required for the translational assay are controlled by the simple operation of two pushbuttons on the device. It is demonstrated that the developed microfluidic device provides precise quantification of two amino acids (methionine and phenylalanine) within 30 min at room temperature. We expect that the application of the presented device can be readily extended to the point-of-care testing of other metabolic compounds.

Pushbutton-activated microfluidic dropenser for droplet digital PCR.

Here, we report a portable microfluidic device to generate and dispense droplets simply operated by pushbutton for droplet digital polymerase chain reaction (ddPCR), which is named pushbutton-activated microfluidic dropenser (droplet dispenser) (PAMD). After loading the PCR mixtures and the droplet generation oil to PAMD, digitized PCR mixtures are prepared in PCR tubes after the actuation of a pushbutton. Multiple droplet generation units are simultaneously operated by a single pushbutton, and the size of droplets is controllable by adjusting the geometry of the droplet generation channel. To examine the performance of PAMD, digitized PCR mixtures containing genomic DNA of Escherichia coli (E. coli) O157:H7 prepared by PAMD were assessed by a fluorescence signal analyzer after PCR with a thermal cycler. As a result, PAMD can produce analytical droplets for ddPCR as much as a conventional droplet generator even though any external equipment is not required.

Reciprocating flow-assisted nucleic acid purification using a finger-actuated microfluidic device.

Molecular diagnostics can provide a powerful diagnostic tool since it can detect pathogens with high sensitivity, but complicated sample preparation procedures limit its widespread use as an on-site detection tool that relies on the skilled person and external equipment. To resolve these limitations, we report a solid-phase nucleic acid purification using a finger-actuated microfluidic device, which can control a set amount of flow regardless of differences in end-users. To increase the recovery rate, a finger-actuated reciprocator was newly developed and integrated into the microfluidic device that can efficiently react with silica microbeads and reagents. After verifying the finger-actuated microfluidic reciprocator, the effect of the reciprocating flow on the recovery rate was assessed to purify the standard DNA of the hepatitis B virus (HBV). The recovery rate was increased up to ∼50% and 955 to 955 000 IU mL-1 of HBV standard DNA was successfully purified and detected by a real-time polymerase chain reaction. Furthermore, the proposed microfluidic device was exploited to purify the HBV DNA from the patient's blood plasma samples.

Towards practical sample preparation in point-of-care testing: user-friendly microfluidic devices.

Microfluidic technologies offer a number of advantages for sample preparation in point-of-care testing (POCT), but the requirement for complicated external pumping systems limits their wide use. To facilitate sample preparation in POCT, various methods have been developed to operate microfluidic devices without complicated external pumping systems. In this review, we introduce an overview of user-friendly microfluidic devices for practical sample preparation in POCT, including self- and hand-operated microfluidic devices. Self-operated microfluidic devices exploit capillary force, vacuum-driven pressure, or gas-generating chemical reactions to apply pressure into microchannels, and hand-operated microfluidic devices utilize human power sources using simple equipment, including a syringe, pipette, or simply by using finger actuation. Furthermore, this review provides future perspectives to realize user-friendly integrated microfluidic circuits for wider applications with the integration of simple microfluidic valves.