Sejung Kim

Rapid Isolation and Detection of Exosomes and Associated Biomarkers from Plasma

Exosomes found in the circulation are a primary source of important cancer-related RNA and protein biomarkers that are expected to lead to early detection, liquid biopsy, and point-of-care diagnostic applications. Unfortunately, due to their small size (50-150 nm) and low density, exosomes are extremely difficult to isolate from plasma. Current isolation methods are time-consuming multistep procedures that are unlikely to translate into diagnostic applications. To address this issue, we demonstrate the ability of an alternating current electrokinetic (ACE) microarray chip device to rapidly isolate and recover glioblastoma exosomes from undiluted human plasma samples. The ACE device requires a small plasma sample (30-50 μL) and is able to concentrate the exosomes into high-field regions around the ACE microelectrodes within 15 min. A simple buffer wash removes bulk plasma materials, leaving the exosomes concentrated on the microelectrodes. The entire isolation process and on-chip fluorescence analysis is completed in less than 30 min which enables subsequent on-chip immunofluorescence detection of exosomal proteins, and provides viable mRNA for RT-PCR analysis. These results demonstrate the ability of the ACE device to streamline the process for isolation and recovery of exosomes, significantly reducing the number of processing steps and time required.

An Aqueous Single Reactor Arc Discharge Process for the Synthesis of Graphene Nanospheres

Using an aqueous single reactor arc discharge process with oil-in-water emulsions allows production of 2D multilayered graphenes (MLGs and 3D graphene-based crumpled/sphere-like particles with low levels of defects). The confinement forces to create 3D particles from 2D MLGs are estimated to be 2.5 μN for crumpled particles and 70 μN for spherical hollow particles.

CTAB enhancement of FRET in DNA structures

The effect of cetyl-trimethylammonium bromide (CTAB) on enhancing the fluorescence resonance energy transfer (FRET) between two dye-conjugated DNA strands was studied using fluorescence emission spectroscopy and dynamic light scattering (DLS). For hybridized DNA where one strand is conjugated with a TAMRA donor and the other with a TexasRed acceptor, increasing the concentration of CTAB changes the fluorescence emission properties and improves the FRET transfer efficiency through changes in the polarity of the solvent, neutralization of the DNA backbone and micelle formation. For the DNA FRET system without CTAB, the DNA hybridization leads to contact quenching between TAMRA donor and TexasRed acceptor producing reduced donor emission and only a small increase in acceptor emission. At 50 µM CTAB, however, the sheathing and neutralization of the dye-conjugated dsDNA structure significantly reduces quenching by DNA bases and dye interactions, producing a large increase in FRET efficiency, which is almost four fold higher than without CTAB.

A Programmable DNA Double-Write Material: Synergy of Photolithography and Self-Assembly Nanofabrication

We demonstrate a DNA double-write process that uses UV to pattern a uniquely designed DNA write material, which produces two distinct binding identities for hybridizing two different complementary DNA sequences. The process requires no modification to the DNA by chemical reagents and allows programmed DNA self-assembly and further UV patterning in the UV exposed and nonexposed areas. Multilayered DNA patterning with hybridization of fluorescently labeled complementary DNA sequences, biotin probe/fluorescent streptavidin complexes, and DNA patterns with 500 nm line widths were all demonstrated.

Influence of MWCNTs on β-Phase PVDF and Triboelectric Properties

The surface of multiwalled carbon nanotubes (MWCNTs) was chemically modified using 1-pyrenebutyric acid (PBA) to improve its compatibility with polyvinylidene fluoride (PVDF). The carboxylic acid groups of the MWCNTs-PBA (PCNTs) provide a β-phase nucleation site to the fluorine of PVDF along their surface. The content of the β-phase crystalline structure of PVDF was found to be the highest at a concentration of 1.0 wt.% of PCNTs, and these PVDF-PCNTs composites were utilized as active layers in triboelectric devices. The maximum output voltage achieved was 16 volts at a concentration of 1.0 wt.% of PCNTs in the PVDF composites.

Polymorphic Architectures of Graphene Quantum Dots

A systematic strategy for designing structured nanomaterials is demonstrated through self-assembly of graphene quantum dots. The approach reveals that graphene derivatives at the nanoscale assemble into various architectures of nanocrystals in a binary solution system. The shapes of the nanocrystals continue to evolve in terms of the intimate association of organic molecules with the dispersion medium, obtaining a high index faceted superlattice. This facile synthetic process provides a versatile strategy for designing particles to new structured materials systems, exploiting the crystallization of layered graphitic carbon structures within single crystals.

DNA multi-bit non-volatile memory and bit-shifting operations using addressable electrode arrays and electric field-induced hybridization

DNA has been employed to either store digital information or to perform parallel molecular computing. Relatively unexplored is the ability to combine DNA-based memory and logical operations in a single platform. Here, we show a DNA tri-level cell non-volatile memory system capable of parallel random-access writing of memory and bit shifting operations. A microchip with an array of individually addressable electrodes was employed to enable random access of the memory cells using electric fields. Three segments on a DNA template molecule were used to encode three data bits. Rapid writing of data bits was enabled by electric field-induced hybridization of fluorescently labeled complementary probes and the data bits were read by fluorescence imaging. We demonstrated the rapid parallel writing and reading of 8 (23) combinations of 3-bit memory data and bit shifting operations by electric field-induced strand displacement. Our system may find potential applications in DNA-based memory and computations.DNA based technology holds promise for non-volatile memory and computational tasks, yet the relatively slow hybridization kinetics remain a bottleneck. Here, Song et al. have developed an electric field-induced hybridization platform that can speed up multi-bit memory and logic operations.

Enhancement of fluorescent resonant energy transfer and the antenna effect in DNA structures with multiple fluorescent dyes

This study examines the use of surfactants and metal cations for the enhancement of long range fluorescent resonant energy transfer (FRET) and the antenna effect in double-stranded (ds) DNA structures formed by hybridization of 21mer oligonucleotides with three fluorescent TAMRA donor dyes and complementary 21mer oligonucleotides with one fluorescent Texas Red acceptor dye. In FRET ds-DNA structures, hydrophobic interactions between the fluorescent dyes in close proximity produces dimerization and quenching which reduces fluorescent emissions. For the neutral surfactant Triton X-100, dimerization and emission quenching in the FRET ds-DNA structures remain unaffected. The cationic surfactant CTAB (>100 μM), which neutralizes the negatively charged ds-DNA backbone reduces TAMRA dye dimerization and emission quenching, and improves the Texas Red quantum yield, FRET efficiency and the antenna effect. While the negatively charged SDS surfactant does not reduce dimerization and emission quenching, addition of sodium cations (Na+) and magnesium cations (Mg2+) lead to a significant reduction in dimerization and emission quenching, and produce higher FRET efficiency and enhanced antenna effect. This study provides a viable strategy for using combinations of surfactants and cations to reduce fluorescent dye and other quenching mechanisms and improve the overall long distance FRET efficiency and the antenna effect in ds-DNA structures.

An Implantable Transparent Conductive Film with Water Resistance and Ultrabendability for Electronic Devices

Recently, instead of indium tin oxide, the random mesh pattern of metallic nanowires for flexible transparent conducting electrodes (FTCEs) has received a great amount of interest due to its flexibility, low resistance, reasonable price, and compliant processes. Mostly, nanowires for FTCEs are fabricated by spray or mayer coating methods. However, the metallic nanowire layer of FTCEs, which is fabricated by these methods, has a spiked surface roughness and low junction contact between the nanowires that lead to their high sheet resistance value. Also, the nanowires on FTCEs are easy to peel-off through exterior forces such as bending, twisting, or contact. To solve these problems, we demonstrate novel methods through which silver nanowires (AgNWs) are deposited onto a nanosize porous nitrocellulose (NC) substrate by electrophoretic deposition (EPD) and an opaque and porous substrate. Respectively, through dimethyl sulfoxide treatment, AgNWs on NC (AgNW/NC) is changed to the transparent and nonporous FTCEs. This enhances the junction contact of the AgNWs by EPD and also allows a permanent attachment of AgNWs onto the substrate. To show the mechanical strength of the AgNWs on the transparent nitrocellulose (AgNW/TNC), it is tested by applying diverse mechanical stress, such as a binding test (3M peel-off), compressing, bending, twisting, and folding. Next, we demonstrate that AgNW/TNC can be effectively implanted onto normal newspapers and papers. As paper electronics, light-emitting diodes, which are laminated onto paper, are successfully operated through a basic AgNW/TNC strip circuit. Finally, it is demonstrated that AgNW/TNC and AgNW/TNC on paper are water resistant for 15 min due to the insulation properties of the nonporous substrate.