Artificial Intelligence-Controlled Microfluidic Device for Fluid Automation and Bubble Removal of Immunoassay Operated by a Smartphone.

There have been tremendous innovations in microfluidic clinical diagnostics to facilitate novel point-of-care testing (POCT) over the past decades. However, the automatic operation of microfluidic devices that minimize user intervention still lacks reliability and repeatability because microfluidic errors such as bubbles and incomplete filling pose a major bottleneck in commercializing the microfluidic devices for clinical testing. In this work, for the first time, various states of microfluid were recognized to control immunodiagnostics by artificial intelligence (AI) technology. The developed AI-controlled microfluidic platform was operated via an Android smartphone, along with a low-cost polymer device to effectuate enzyme-linked immunosorbent assay (ELISA). To overcome the limited machine-learning capability of smartphones, the region-of-interest (ROI) cascading and conditional activation algorithms were utilized herein. The developed microfluidic chip was incorporated with a bubble trap to remove any bubbles detected by AI, which helps in preventing false signals during immunoassay, as well as controlling the reagents' movement with an on-chip micropump and valve. Subsequently, the developed immunosensing platform was tested for conducting real ELISA using a single microplate from the 96-well to detect the Human Cardiac Troponin I (cTnI) biomarker, with a detection limit as low as 0.98 pg/mL. As a result, the developed platform can be envisaged as an AI-based revolution in microfluidics for point-of-care clinical diagnosis.

Smartphone-Based Fully Automated Optofluidic Device with Laser Irradiation-Induced Image Whitening.

Herein, we have developed a fully automated optofluidic device to execute enzyme-linked immunosorbent assay (ELISA) using an active 96-well hybrid lab-on-a-chip (LOC) device. To automate the solution loading into the reaction zone of the device and the post-assayed signal analysis, laser irradiation-induced image whitening was utilized with a smartphone-based optical platform. Two optical phenomena were utilized in our platform to detect the liquid in the reaction chamber using a smartphone. First, by Fresnel's equation, the refraction difference between air and water resulted in the intensity change of the reflected light from the reaction chamber. Therefore, when the liquid was entering into the reaction chamber, the intensity of the reflected light was changed. Second, when the light intensity increases, the smartphone-captured image whitens out due to saturation, even when the red color light was incident. Therefore, by measuring the RGB value of the smartphone image, the intensity changes by the liquid movement in the reaction chamber were successfully monitored. Our platform showed a low detection limit of 7.81 pg/mL for the detection of the NT-proBNP human cardiac biomarker with almost a half standard deviation, compared to the manually operated LOC-based ELISA. As a fully automated LOC adopting a conventional 96-well ELISA platform, we thus concluded that the developed platform can be widely applied for point-of-care clinical tests.

Microfluidic Adapter Converting a 96-Well Cartridge into an Autonomous Microfluidic Device.

In this study, an immunosensing platform by integrating a single array of conventional 96-well cartridges with low-cost polydimethylsiloxane (PDMS) pillars and thermoplastic chip was designed and developed to execute enzyme-linked immunosorbent assay (ELISA). The platform provides multiple (eight) reaction chambers for sequentially detecting multiple analytes under similar assay environments. For multiple immunoassays, the reaction chambers are consequently activated using a microvalve integrated with the sensing zones via the thermoplastic chip. Because the reaction zones are able to be selected and isolated from each other by the microvalve, multiple immunoassays can be implemented, avoiding cross-contamination. The performance of the sensing platform demonstrated its effectiveness in assaying with an optimal sample volume of 10 µL with an assay time of 10-15 min for different assay steps, which is much lower than the conventional immunoassay using 96-well plates. Thus, a low detection limit of 9.75 pg/mL is achieved using the developed platform to successfully detect the cardiac troponin I (cTnI). As a result, the cost-effectively made PDMS pillars and 96-well cartridge based hybrid immunosensing platform are widely applicable for high-throughput multiple lab-on-a-chip immunoassays.

Paper-Plastic Hybrid Microfluidic Device for Smartphone-Based Colorimetric Analysis of Urine.

In this work, a disposable paper-plastic hybrid microfluidic lab-on-a-chip (LOC) has been developed and successfully applied for the colorimetric measurement of urine by the smartphone-based optical platform using a "UrineAnalysis" Android app. The developed device was cost-effectively implemented as a stand-alone hybrid LOC by incorporating the paper-based conventional reagent test strip inside the plastic-based LOC microchannel. The LOC device quantitatively investigated the small volume (40 µL) of urine analytes for the colorimetric reaction of glucose, protein, pH, and red blood cell (RBC) in integration with the finger-actuating micropump. On the basis of our experiments, the conventional urine strip showed large deviation as the reaction time goes by, because dipping the strip sensor in a bottle of urine could not control the reaction volume. By integrating the strip sensor in the LOC device for urine analysis, our device significantly improves the time-dependent inconstancy of the conventional dipstick-based urine strip, and the smartphone app used for image analysis enhances the visual assessment of the test strip, which is a major user concern for the colorimetric analysis in point-of-care (POC) applications. As a result, the user-friendly LOC, which is successfully implemented in a disposable format with the smartphone-based optical platform, may be applicable as an effective tool for rapid and qualitative POC urinalysis.

A smart pipette for equipment-free separation and delivery of plasma for on-site whole blood analysis.

A novel device of smart pipette has been suggested to extract and deliver plasma from whole blood in a disposable format. By operating an on-chip disposable micropump, approximately 30 µL of plasma was obtained from 100 µL of whole blood within 5 min without any external equipment for point-of-care blood analysis.

Polymer-Supported Graphene Sheet as a Vertically Conductive Anode of Lithium-Ion Battery.

The increasing demand for electric vehicles necessitates the development of cost-effective, mass-producible, long-lasting, and highly conductive batteries. Making this kind of battery is exceedingly tricky. This study introduces an innovative fabrication technique utilizing a laser-induced graphene (LIG) approach on commercial Kapton film to create hexagonal pores. These pores form vertical conduction paths for electron and ion transportation during lithiation and delithiation, significantly enhancing conductivity. The nongraphitized portion of the Kapton film makes it a binder-less, free-standing electrode, providing mechanical stability. Various analytical techniques, including scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), Raman spectroscopy, and atomic force microscopy (AFM) are utilized to confirm the transformation of a 3D porous graphene sheet from a commercial Kapton film. Cross-sectional SEM images verify the vertical connections. The specific capacity of 581 mAh g-1 is maintained until the end, with 99% coulombic efficiency at 0.1C. This simple manufacturing method paves the pathway for future LIG-based, cost-effective, lightweight, mass-producible, long-lasting, vertically conductive electrodes for lithium-ion batteries.

Formation of lipid bilayers inside microfluidic channel array for monitoring membrane-embedded nanopores of phi29 DNA packaging nanomotor.

An efficient method to form lipid bilayers inside an array of microfluidic channels has been developed and applied to monitor the membrane-embedded phi29 DNA packaging motor with an electrochemical characterization on a lab-on-a-chip (LOC) platform. A push-pull junction capturing approach was applied to confine a small amount of the lipid solution inside a microchannel. The selective permeability between solvents and water in PDMS was utilized to extract the solvent from the lipid solution, resulting in a self-formation of the lipid bilayer in the microchannel array. Each microchannel was independently connected to a silver/silver chloride (Ag/AgCl) electrode array, leading to a high-throughput monitoring of the nanopore insertion in the formed lipid bilayers. The formation of multiple lipid bilayers inside an array of microchannels and the simultaneous electrical and optical monitoring of multiple bilayer provides an efficient LOC platform for the further development of single phi29 motor pore sensing and high throughput single pore dsDNA sequencing.

Fully integrated rapid microfluidic device translated from conventional 96-well ELISA kit.

In this work, a fully integrated active microfluidic device transforming a conventional 96-well kit into point-of-care testing (POCT) device was implemented to improve the performance of traditional enzyme-linked immunosorbent assay (ELISA). ELISA test by the conventional method often requires the collection of 96 samples for its operation as well as longer incubation time from hours to overnight, whereas our proposed device conducts ELISA immediately individualizing a 96-well for individual patients. To do that, a programmable and disposable on-chip pump and valve were integrated on the device for precise control and actuation of microfluidic reagents, which regulated a reaction time and reagent volume to support the optimized protocols of ELISA. Due to the on-chip pump and valve, ELISA could be executed with reduced consumption of reagents and shortening the assay time, which are crucial for conventional ELISA using 96-well microplate. To demonstrate highly sensitive detection and easy-to-use operation, this unconventional device was successfully applied for the quantification of cardiac troponin I (cTnI) of 4.88 pg/mL using a minimum sample volume of 30 µL with a shorter assay time of 15 min for each ELISA step. The limit of detection (LOD) thus obtained was significantly improved than the conventional 96-well platform.

An on-chip whole blood/plasma separator with bead-packed microchannel on COC polymer.

A disposable on-chip whole blood/plasma separator, which is able to separate plasma from whole human blood by capillary force through a bead-packed microchannel, has been designed, fabricated and characterized in this work. Various sizes of silica beads were slurry-packed through a microchannel using a bump structure which held beads in a defined region. The bead-packed microchannel induces a capillary force which allows plasma to move forward through the bead-packed column more rapidly than red blood cells (RBCs). The blood/plasma separator with bead-packed microchannel has successfully separated plasma from the whole blood without haemolysis of RBCs. The separation method developed in this work can be applied to various on-chip stationary filtrations of RBC for point-of-care clinical diagnostics.

Self-aligned nanogaps on multilayer electrodes for fluidic and magnetic assembly of carbon nanotubes.

A self-aligned nanogap between multiple metal layers has been developed using a new controlled undercut and metallization technique (CUMT), and practically applied for self-assembly of individual carbon nanotubes (CNTs) over the developed nanogap. This new method allows conventional optical lithography to fabricate nanogap electrodes and self-aligned patterns with nanoscale precision. The self-aligned nickel (Ni) pattern on the nanogap electrode works as an assembly spot where the residual iron (Fe) catalyst at the end of the CNT is magnetically captured. The captured CNT is forced to be aligned parallel to the flow direction by fluidic shear force. The combined forces of magnetic attraction and fluidic alignment provide massive self-assembly of CNTs at target positions. Both multiwalled nanotubes (MWNTs) and single walled nanotubes (SWNTs) were successfully assembled over the nanogap electrodes, and their electrical characteristics were fully characterized. The CNTs self-assembled on the developed electrodes with a nanogap and showed a very reliable and reproducible current-voltage (I-V) characteristic. The method developed in this work can envisage the mass fabrication of individual CNT-assembled devices which can be applied to nanoelectronic devices or nanobiosensors.

Toward a disposable low-cost LOC device: heterogeneous polymer micro valve and pump fabricated by UV/ozone-assisted thermal fusion bonding.

Herein, a heterogeneous polymer micro valve and pump with a polypropylene (PP) membrane was developed in a low-cost manner via UV/ozone-assisted thermal fusion bonding. The proposed fabrication technique allowed for a geometrically selective bonding; consequently, the membrane was prevented from bonding with the valve seat of the diaphragm micro-valve, without patterning a protection layer or introducing an additional structure. The developed device withstands 480 kPa of static pressure and up to 350 kPa of a vibration pressure, providing sufficient bonding strength for microfluidic actuations. The fabricated micro valve and pump are fully characterized and compared with a poly(dimethylsiloxane) (PDMS) membrane glass device, showing comparable valving and pumping performance. As a result, the robust PP membrane micro valve and pump are simply implemented in a facile manner, and demonstrated excellent performance, which is highly desirable for mass production of disposable lab-on-a-chip (LOC) devices.

The precise self-assembly of individual carbon nanotubes using magnetic capturing and fluidic alignment.

A new method for the self-assembly of a carbon nanotube (CNT) using magnetic capturing and fluidic alignment has been developed and characterized in this work. In this new method, the residual iron (Fe) catalyst positioned at one end of the CNT was utilized as a self-assembly driver to attract and position the CNT, while the assembled CNT was aligned by the shear force induced from the fluid flow through the assembly channel. The self-assembly procedures were successfully developed and the electrical properties of the assembled multi-walled carbon nanotube (MWNT) and single-walled carbon nanotube (SWNT) were fully characterized. The new assembly method developed in this work shows its feasibility for the precise self-assembly of parallel CNTs for electronic devices and nanobiosensors.

A disposable on-chip microvalve and pump for programmable microfluidics.

In this work, a low-cost PDMS micro-pump and -valve have been designed and developed to control multiple reagents for enzyme-linked immunosorbent assay (ELISA) on a programmable lab-on-a-chip (LOC) platform. The micro pump and valves were precisely controlled by selectively pressurizing the PDMS channels and chamber to actuate the multiple reagents in a controlled manner. Selective pressurizing of the PDMS structures was initiated by a simple system that maneuvered a single roller bar operated by a programmed microprocessor. The performance of the micro-pump was fully characterized and a minimum fluid volume of 1 µL was controlled. Also, the on-chip microvalves were programmed to flow the multiple reagents to automatically process the multi-step ELISA procedures. By applying the proposed platform, 19.40 pg ml-1 cardiac troponin T (cTnT) was successfully detected on the LOC device by using multiple programmed valves as multiple steps of the enzyme-linked sandwich immunoassay. As a result, the developed micro-pump and -valve, which were successfully applied to actuate a series of solutions in a controlled manner, can be widely applied to lab-on-a-chip based bioassays.

On-chip signal amplification of magnetic bead-based immunoassay by aviating magnetic bead chains.

In this work, a Lab-on-a-Chip (LOC) platform is used to electromagnetically actuate magnetic bead chains for an enhanced immunoassay. Custom-made electromagnets generate a magnetic field to form, rotate, lift and lower the magnetic bead chains (MBCs). The cost-effective, disposable LOC platform was made with a polymer substrate and an on-chip electrochemical sensor patterned via the screen-printing process. The movement of the MBCs is controlled to improve the electrochemical signal up to 230% when detecting beta-type human chorionic gonadotropin (ß-hCG). Thus, the proposed on-chip MBC-based immunoassay is applicable for rapid, qualitative electrochemical point-of-care (POC) analysis.

Histogram analysis for smartphone-based rapid hematocrit determination.

A novel and rapid analysis technique using histogram has been proposed for the colorimetric quantification of blood hematocrits. A smartphone-based "Histogram" app for the detection of hematocrits has been developed integrating the smartphone embedded camera with a microfluidic chip via a custom-made optical platform. The developed histogram analysis shows its effectiveness in the automatic detection of sample channel including auto-calibration and can analyze the single-channel as well as multi-channel images. Furthermore, the analyzing method is advantageous to the quantification of blood-hematocrit both in the equal and varying optical conditions. The rapid determination of blood hematocrits carries enormous information regarding physiological disorders, and the use of such reproducible, cost-effective, and standard techniques may effectively help with the diagnosis and prevention of a number of human diseases.

Optical immunosensor using carbon nanotubes coated with a photovoltaic polymer.

In this work, an on-chip optical immunosensor using an individually assembled carbon nanotube (CNT) coated with a photovoltaic polymer has been proposed, developed, characterized, and applied for the detection of cardiac biomarkers. An individual CNT was self-assembled on a nickel (Ni)-patterned electrode by magnetically attracting the residual iron catalyst at one end of the CNT. After the CNT self-assembled electrode was prepared, it was coated with a photovoltaic polymer to implement a CNT photodetector. Under an incident light, the photovoltaic polymer generated electrons that changed the conductivity of the CNT. The CNT photodetector was finally insulated with parylene to prevent interruptions of charged molecules in a sample solution, such as non-specifically bound proteins and various ions. Chemiluminescent immunoassay was directly performed on the CNT photodetector for an on-chip detection of cardiac troponin T (cTnT) with a detection limit of 12 pg/mL. High sensitivity and reliable selectivity have been achieved through the use of on-chip measurement of chemiluminescent light by the CNT photodetector. As a result, the developed device is envisaged as a new platform for optical immunosensing using the individually self-assembled CNT for point-of-care (POC) clinical diagnostics.

An on-chip whole blood/plasma separator using hetero-packed beads at the inlet of a microchannel.

A disposable rapid on-chip whole blood/plasma separator has been simply implemented by packing two different sizes of silica beads at the inlet of a microchannel for application to point-of-care (POC) clinical diagnostics. Our suggested technique can be utilized to implement an integrated microfilter for a wide range of Lab-on-a-Chip (LOC) devices.