Whole Genome Sequencing of Single Circulating Tumor Cells Isolated by Applying a Pulsed Laser to Cell-Capturing Microstructures.

Single cell analysis of heterogeneous circulating tumor cells (CTCs), by which the genomic profiles of rare single CTCs are connected to the clinical status of cancer patients, is crucial for understanding cancer metastasis and the clinical impact on patients. However, the heterogeneity in genotypes and phenotypes and rarity of CTCs have limited extensive single CTC genome research, further hindering clinical investigation. Despite recent efforts to build platforms that separate CTCs, the investigation on CTCs is difficult due to the lack of a retrieval process at the single cell level. In this study, laser-induced isolation of microstructures on an optomechanically-transferrable-chip and sequencing (LIMO-seq) is applied for whole genome sequencing of single CTCs. Also, the whole genome sequences and the molecular profiles of the isolated single cells from the whole blood of a breast cancer patient are analyzed.

Gradient-Wrinkled Microparticle with Grayscale Lithography Controlling the Cross-Linking Densities for High Security Level Anti-Counterfeiting Strategies.

Physical unclonable functions (PUFs) enable different characteristics according to the purpose, such as easy to access identification, high security level, and high code capacity, against counterfeiting a product. However, most multiplex approaches have been implemented by embedding several security features rather than one feature. In this paper, we present a high security level anti-counterfeiting strategy using only labyrinth wrinkle patterns with different complexities, which can be used as unique and unclonable codes. To generate codes with different levels in a microtaggant, we fabricated wrinkle patterns with characteristic wavelength gradients using grayscale lithography. The elastic modulus of the polymer substrate and corresponding wavelength after the wrinkling process were controlled by designing the gray level of each subcode region in a gray-level mask image for photopolymerization of the microparticle substrate. We then verified the uniqueness of the extracted minutia codes through a cross-correlation analysis. Finally, we demonstrated the authentication strategies by decoding different minutia codes according to the scanning resolution during the decoding. Overall, the presented patterning method can be widely used in security code generation.

DNA Micro-Disks for the Management of DNA-Based Data Storage with Index and Write-Once-Read-Many (WORM) Memory Features.

DNA-based data storage has attracted attention because of its higher physical density of the data and longer retention time than those of conventional digital data storage. However, previous DNA-based data storage lacked index features and the data quality of storage after a single access was not preserved, obstructing its industrial use. Here, DNA micro-disks, QR-coded micro-sized disks that harbor data-encoded DNA molecules for the efficient management of DNA-based data storage, are proposed. The two major features that previous DNA-based data-storage studies could not achieve are demonstrated. One feature is accessing data items efficiently by indexing the data-encoded DNA library. Another is achieving write-once-read-many (WORM) memory through the immobilization of DNA molecules on the disk and their enrichment through in situ DNA production. Through these features, the reliability of DNA-based data storage is increased by allowing selective and multiple accession of data-encoded DNA with lower data loss than previous DNA-based data storage methods.

Curved neuromorphic image sensor array using a MoS2-organic heterostructure inspired by the human visual recognition system.

Conventional imaging and recognition systems require an extensive amount of data storage, pre-processing, and chip-to-chip communications as well as aberration-proof light focusing with multiple lenses for recognizing an object from massive optical inputs. This is because separate chips (i.e., flat image sensor array, memory device, and CPU) in conjunction with complicated optics should capture, store, and process massive image information independently. In contrast, human vision employs a highly efficient imaging and recognition process. Here, inspired by the human visual recognition system, we present a novel imaging device for efficient image acquisition and data pre-processing by conferring the neuromorphic data processing function on a curved image sensor array. The curved neuromorphic image sensor array is based on a heterostructure of MoS2 and poly(1,3,5-trimethyl-1,3,5-trivinyl cyclotrisiloxane). The curved neuromorphic image sensor array features photon-triggered synaptic plasticity owing to its quasi-linear time-dependent photocurrent generation and prolonged photocurrent decay, originated from charge trapping in the MoS2-organic vertical stack. The curved neuromorphic image sensor array integrated with a plano-convex lens derives a pre-processed image from a set of noisy optical inputs without redundant data storage, processing, and communications as well as without complex optics. The proposed imaging device can substantially improve efficiency of the image acquisition and recognition process, a step forward to the next generation machine vision.

OPENchip: an on-chip in situ molecular profiling platform for gene expression analysis and oncogenic mutation detection in single circulating tumour cells.

Liquid biopsy holds promise towards practical implementation of personalized theranostics of cancer. In particular, circulating tumour cells (CTCs) can provide clinically actionable information that can be directly linked to prognosis or therapy decisions. In this study, gene expression patterns and genetic mutations in single CTCs are simultaneously analysed by strategically combining microfluidic technology and in situ molecular profiling technique. Towards this, the development and demonstration of the OPENchip (On-chip Post-processing ENabling chip) platform for single CTC analysis by epithelial CTC enrichment and subsequent in situ molecular profiling is reported. For in situ molecular profiling, padlock probes that identify specific desired targets to examine biomarkers of clinical relevance in cancer diagnostics were designed and used to create libraries of rolling circle amplification products. We characterize the OPENchip in terms of its capture efficiency and capture purity, and validate the probe design using different cell lines. By integrating the obtained results, molecular analyses of CTCs from metastatic breast cancer (HER2 (ERBB2) gene expression and PIK3CA mutations) and metastatic pancreatic cancer (KRAS gene mutations) patients were demonstrated without any off-chip processes. The results substantiate the potential implementation of early molecular detection of cancer through sequencing-free liquid biopsy.

One-Step Generation of a Drug-Releasing Hydrogel Microarray-On-A-Chip for Large-Scale Sequential Drug Combination Screening.

Large-scale screening of sequential drug combinations, wherein the dynamic rewiring of intracellular pathways leads to promising therapeutic effects and improvements in quality of life, is essential for personalized medicine to ensure realistic cost and time requirements and less sample consumption. However, the large-scale screening requires expensive and complicated liquid handling systems for automation and therefore lowers the accessibility to clinicians or biologists, limiting the full potential of sequential drug combinations in clinical applications and academic investigations. Here, a miniaturized platform for high-throughput combinatorial drug screening that is "pipetting-free" and scalable for the screening of sequential drug combinations is presented. The platform uses parallel and bottom-up formation of a heterogeneous drug-releasing hydrogel microarray by self-assembly of drug-laden hydrogel microparticles. This approach eliminates the need for liquid handling systems and time-consuming operation in high-throughput large-scale screening. In addition, the serial replacement of the drug-releasing microarray-on-a-chip facilitates different drug exchange in each and every microwell in a simple and highly parallel manner, supporting scalable implementation of multistep combinatorial screening. The proposed strategy can be applied to various forms of combinatorial drug screening with limited amounts of samples and resources, which will broaden the use of the large-scale screening for precision medicine.

Hierarchical shape-by-shape assembly of microparticles for micrometer-scale viral delivery of two different genes.

Understanding tissue engineering using a bottom-up approach has been hindered by technical limitations because no platform can demonstrate the controlled formation of a heterogeneous population of cells in microscale. Here, we demonstrate hierarchical shape-by-shape assembly of virus-laden particles into larger ones to transfect two different genes on the seeded cells. We show that smaller daughter particles with different sizes and shapes can be assembled into the matching indentations of larger parent particles with different sizes and shapes. Then, we transfected a population of cells with two different gene-transfecting viruses, each of which was laden on the parent or daughter particles.

An encoded viral micropatch for multiplex cell-based assays through localized gene delivery.

The increasing number of potential drug targets and compounds has led to the development of high-throughput cell-based assays. Simultaneous processing of multiple targets in the same experiment based on localized target gene expression is a very efficient strategy for this purpose. To address this need, we present an adenoviral vector-immobilized microparticle with two-dimensional (2D) shape-encoding properties that allows localized patch-like gene delivery to monolayer-cultured cells. This format conveniently achieves multiplexed gene delivery compatible with both high-throughput cellular assays and fluorescence high-content imaging instruments. A multiplex G protein-coupled receptor (GPCR) internalization assay was developed to demonstrate the compatibility of this system with high-throughput image-based cellular assays.

Self-organization of maze-like structures via guided wrinkling.

Sophisticated three-dimensional (3D) structures found in nature are self-organized by bottom-up natural processes. To artificially construct these complex systems, various bottom-up fabrication methods, designed to transform 2D structures into 3D structures, have been developed as alternatives to conventional top-down lithography processes. We present a different self-organization approach, where we construct microstructures with periodic and ordered, but with random architecture, like mazes. For this purpose, we transformed planar surfaces using wrinkling to directly use randomly generated ridges as maze walls. Highly regular maze structures, consisting of several tessellations with customized designs, were fabricated by precisely controlling wrinkling with the ridge-guiding structure, analogous to the creases in origami. The method presented here could have widespread applications in various material systems with multiple length scales.

Shape-encoded silica microparticles for multiplexed bioassays.

Shape-encoded silica microparticles for use in multiplexed bioassays were fabricated by using optofluidic maskless lithography (OFML) and tetraethylorthosilicate (TEOS) polymerization. These encoded silica microparticles exhibit excellent bioconjugation properties and negligible non-specific analyte adsorption. Encoded silica microparticles could be useful in a wide variety of applications, including DNA- and protein-based diagnostics.

Biomimetic microfingerprints for anti-counterfeiting strategies.

An unclonable, fingerprint-mimicking anti-counterfeiting strategy is presented that encrypts polymeric particles with randomly generated silica film wrinkles. The generated wrinkle codes are as highly unique as human fingerprints and are technically irreproducible. Superior to previous physical unclonable functions, codes are tunable on demand and generable on various geometries. Reliable authentication of real-world products that have these microfingerprints is demonstrated using optical decoding methods.

Lithographically encoded polymer microtaggant using high-capacity and error-correctable QR code for anti-counterfeiting of drugs.

A QR-coded microtaggant for the anti-counterfeiting of drugs is proposed that can provide high capacity and error-correction capability. It is fabricated lithographically in a microfluidic channel with special consideration of the island patterns in the QR Code. The microtaggant is incorporated in the drug capsule ("on-dose authentication") and can be read by a simple smartphone QR Code reader application when removed from the capsule and washed free of drug.