Rapid detection of pathogens and analytes at the point of care offers an opportunity for prompt patient management and public health control. This paper reports an open microfluidic platform coupled with active whispering gallery mode (WGM) microsphere resonators for the rapid detection of influenza viruses. The WGM microsphere resonators, precoated with influenza A polyclonal antibodies, are mechanically trapped in the open micropillar array, where the evaporation-driven flow continuously transports a small volume (∼μL) of sample to the resonators without auxiliaries. Selective chemical modification of the pillar array changes surface wettability and flow pattern, which enhances the detection sensitivity of the WGM resonator-based virus sensor. The optofluidic sensing platform is able to specifically detect influenza A viruses within 15 min using a few microliters of sample and displays a linear response to different virus concentrations.

Chimeric antigen receptor (CAR)-T cell therapy has emerged as a promising cell-based immunotherapy approach for treating blood disorders and cancers, but genetically engineering CAR-T cells is challenging due to primary T cells' sensitivity to conventional gene delivery approaches. The current viral-based method can typically involve significant operating costs and biosafety hurdles, while bulk electroporation (BEP) can lead to poor cell viability and functionality. Here, a non-viral electroactive nanoinjection (ENI) platform is developed to efficiently negotiate the plasma membrane of primary human T cells via vertically configured electroactive nanotubes, enabling efficient delivery (68.7%) and expression (43.3%) of CAR genes in the T cells, with minimal cellular perturbation (>90% cell viability). Compared to conventional BEP, the ENI platform achieves an almost threefold higher CAR transfection efficiency, indicated by the significantly higher reporter GFP expression (43.3% compared to 16.3%). By co-culturing with target lymphoma Raji cells, the ENI-transfected CAR-T cells' ability to effectively suppress lymphoma cell growth (86.9% cytotoxicity) is proved. Taken together, the results demonstrate the platform's remarkable capacity to generate functional and effective anti-lymphoma CAR-T cells. Given the growing potential of cell-based immunotherapies, such a platform holds great promise for ex vivo cell engineering, especially in CAR-T cell therapy.

Background Nanoinjection—the process of intracellular delivery using vertically configured nanostructures—is a physical route that efficiently negotiates the plasma membrane, with minimal perturbation and toxicity to the cells. Nanoinjection, as a physical membrane-disruption-mediated approach, overcomes challenges associated with conventional carrier-mediated approaches such as safety issues (with viral carriers), genotoxicity, limited packaging capacity, low levels of endosomal escape, and poor versatility for cell and cargo types. Yet, despite the implementation of nanoinjection tools and their assisted analogues in diverse cellular manipulations, there are still substantial challenges in harnessing these platforms to gain access into cell interiors with much greater precision without damaging the cell’s intricate structure. Here, we propose a non-viral, low-voltage, and reusable electroactive nanoinjection (ENI) platform based on vertically configured conductive nanotubes (NTs) that allows for rapid influx of targeted biomolecular cargos into the intracellular environment, and for successful gene silencing. The localization of electric fields at the tight interface between conductive NTs and the cell membrane drastically lowers the voltage required for cargo delivery into the cells, from kilovolts (for bulk electroporation) to only ≤ 10 V; this enhances the fine control over membrane disruption and mitigates the problem of high cell mortality experienced by conventional electroporation. Results Through both theoretical simulations and experiments, we demonstrate the capability of the ENI platform to locally perforate GPE-86 mouse fibroblast cells and efficiently inject a diverse range of membrane-impermeable biomolecules with efficacy of 62.5% (antibody), 55.5% (mRNA), and 51.8% (plasmid DNA), with minimal impact on cells’ viability post nanoscale-EP (> 90%). We also show gene silencing through the delivery of siRNA that targets TRIOBP, yielding gene knockdown efficiency of 41.3%. Conclusions We anticipate that our non-viral and low-voltage ENI platform is set to offer a new safe path to intracellular delivery with broader selection of cargo and cell types, and will open opportunities for advanced ex vivo cell engineering and gene silencing. Graphical abstract

Cell-based immunotherapy such as chimeric antigen receptor (CAR)-T therapy holds great promise in treating cancer and other diseases; but the current viral-based method represents a significant cost and safety hurdle. Here, we show for the first time successful CAR transfection into primary T cells via vertically aligned silicon nanotube (SiNT) arrays. SiNT-mediated transfection achieves comparable or even higher delivery efficiency (20–37%) and expression efficiency (18–24%) to that achieved by electroporation. Scanning electron microscopy imaging after focused ion beam milling demonstrated the tight T cell–SiNT interface. The induced membrane invaginations and the proximity between individual SiNTs and the nucleus might enhance endocytic pathways, and enable direct delivery of CAR construct into the nucleus, thus resulting in higher CAR expression efficiency. SiNT-interfacing also results in faster proliferation of T cells compared to cells transfected by electroporation; non-activated T (N_SiNT) cells undergo higher numbers of cell division than pre-activated ones (A_SiNT). By co-culturing with target lymphoma Raji cells, we prove that SiNT-transfected CAR-T cells can suppress Raji cell growth, indicated by significant increase in effector:target (E:T) ratio (by up to 30.7-fold). While SiNTs induce an overall upregulation of cytokine production in T cells, N_SiNT T cells exhibited high increase in secretion of IFNγ and IL-6, and relatively high in TNFα, which could contribute to their enhanced killing ability (∼96% cytotoxicity), demonstrated by their stronger inhibition on target Raji cells through luciferase assay. The results demonstrate the capacity of SiNT-mediated transfection of generating effective anti-lymphoma CAR-T cells. Considering the growing potential of cell-based therapies, we expect that a non-viral nanoinjection platform such as ours will facilitate the full realization of their therapeutic promise.

Optofluidic biosensors offer significant advantages for lab-on-a-chip chemical and biological analysis by combining microfluidics for small-volume liquid manipulation with the high sensitivity and adaptability of optical sensing techniques. In this study, we present an optofluidic biosensing platform utilising a micropillar array coupled with active whispering gallery mode (WGM) microsphere resonators for the rapid detection of influenza viruses. The WGM microspheres are coated with polyclonal antibodies capable of targeting both intact virus and fragments of viral proteins. The micropillar array facilitates continuous sampling and measurement through evaporation-driven fluid transport.

Nanoinjection—the process of intracellular delivery using vertically configured nanostructures—is a physical route that efficiently negotiates the plasma membrane, with minimal perturbation and toxicity to the cells. Despite the implementation of nanoinjection tools and their assisted analogues in diverse cellular manipulations, there are still substantial challenges in harnessing these platforms to gain access into cell interiors with much greater precision without damaging the cell’s intricate structure. Here, we propose a low-voltage, simple, scalable, and reusable electroactive nanoinjection (ENI) platform based on vertically configured conductive nanotubes (NTs) that allows for rapid influx of targeted biomolecular cargos into the intracellular environment, and for successful gene editing. The localization of electric fields at the tight interface between conductive NTs and the cell membrane drastically lowers the voltage required for cargo delivery into the cells, from kilovolts (for bulk electroporation) to only ≤10 V; this enhances the fine control over membrane disruption and mitigates the problem of high cell mortality experienced by conventional electroporation. Through both theoretical simulations and experiments, we demonstrate the capability of the ENI platform to locally perforate GPE-86 mouse fibroblast cells and efficiently inject a diverse range of membrane-impermeable biomolecules with efficacy of ~63% (antibody), ~56% (mRNA), and ~52% (plasmid DNA), with minimal impact on cells’ viability post nanoscaleEP (>90%). We also show gene editing through the delivery of siRNA that targets TRIOBP, yielding gene knockdown efficiency of ~41%. We anticipate that our ENI platform is set to offer a new path to intracellular delivery with broader selection of cargo and cell types, and will open opportunities for advanced ex vivo cell engineering and gene editing.

Abstract Nanofabrication technologies have been recently applied to the development of engineered nano–bio interfaces for manipulating complex cellular processes. In particular, vertically configurated nanostructures such as nanoneedles (NNs) have been adopted for a variety of biological applications such as mechanotransduction, biosensing, and intracellular delivery. Despite their success in delivering a diverse range of biomolecules into cells, the mechanisms for NN-mediated cargo transport remain to be elucidated. Recent studies have suggested that cytoskeletal elements are involved in generating a tight and functional cell–NN interface that can influence cargo delivery. In this study, by inhibiting actin dynamics using two drugs—cytochalasin D (Cyto D) and jasplakinolide (Jas), we demonstrate that the actin cytoskeleton plays an important role in mRNA delivery mediated by silicon nanotubes (SiNTs). Specifically, actin inhibition 12 h before SiNT-cellular interfacing (pre-interface treatment) significantly dampens mRNA delivery (with efficiencies dropping to 17.2% for Cyto D and 33.1% for Jas) into mouse fibroblast GPE86 cells, compared to that of untreated controls (86.9%). However, actin inhibition initiated 2 h after the establishment of GPE86 cell–SiNT interface (post-interface treatment), has negligible impact on mRNA transfection, maintaining > 80% efficiency for both Cyto D and Jas treatment groups. The results contribute to understanding potential mechanisms involved in NN-mediated intracellular delivery, providing insights into strategic design of cell–nano interfacing under temporal control for improved effectiveness.

Si–As–Se based OTS materials are studied, the increasing content of silicon leads to a larger threshold voltage and smaller off-state current.

Engineered nano–bio cellular interfaces driven by vertical nanostructured materials are set to spur transformative progress in modulating cellular processes and interrogations. In particular, the intracellular delivery—a core concept in fundamental and translational biomedical research—holds great promise for developing novel cell therapies based on gene modification. This study demonstrates the development of a mechanotransfection platform comprising vertically aligned silicon nanotube (VA‐SiNT) arrays for ex vivo gene editing. The internal hollow structure of SiNTs allows effective loading of various biomolecule cargoes; and SiNTs mediate delivery of those cargoes into GPE86 mouse embryonic fibroblasts without compromising their viability. Focused ion beam scanning electron microscopy (FIB‐SEM) and confocal microscopy results demonstrate localized membrane invaginations and accumulation of caveolin‐1 at the cell–NT interface, suggesting the presence of endocytic pits. Small‐molecule inhibition of endocytosis suggests that active endocytic process plays a role in the intracellular delivery of cargo from SiNTs. SiNT‐mediated siRNA intracellular delivery shows the capacity to reduce expression levels of F‐actin binding protein (Triobp) and alter the cellular morphology of GPE86. Finally, the successful delivery of Cas9 ribonucleoprotein (RNP) to specifically target mouse Hprt gene is achieved. This NT‐enhanced molecular delivery platform has strong potential to support gene editing technologies.

We have successfully developed a mass-productive sputtering module for insulating materials and advanced PbZrTiO3 (PZT) process technology which enable low temperature crystallization of PZT (<500 deg.C) and achieve higher piezoelectric coefficient and breakdown voltage of PZT films

Microfluidic flow in lab-on-a-chip devices is typically very sensitive to the variable physical properties of complex samples, e.g., biological fluids. Here, evaporation-driven fluid transport (transpiration) is achieved in a configuration that is insensitive to interfacial tension, salinity, and viscosity over a wide range. Micropillar arrays ("pillar cuvettes") were preloaded by wicking a known volatile fluid (water) and then adding a microliter sample of salt, surfactant, sugar, or saliva solution to the loading zone. As the preloaded fluid evaporates, the sample is reliably drawn from a reservoir through the pillar array at a rate defined by the evaporation of the preloaded fluid (typically nL/s). Including a reagent in the preloaded fluid allows photometric reactions to take place at the boundary between the two fluids. In this configuration, a photometric signal enhancement is observed and chemical analysis is independent of both humidity and temperature. The ability to reliably transport and sense an analyte in microliter volumes without concern over salt, surfactant, viscosity (in part), humidity, and temperature is a remarkable advantage for analytical purposes.

Engineered nano-bio interfaces driven by tunable vertically configured nanostructures have recently emerged as a powerful tool for cellular manipulations and interrogations. Yet the interplay between substrate topography and cellular behavior is highly complex and not fully understood. A new experimental design is proposed that enables generation of ultrathin sections (lamellae) of cell-nanostructure imprints with minimal artifacts. We demonstrate the potential of such lamellae for efficient transmission electron microscopy (TEM) characterization of interfacial interactions between adherent cells and vertically aligned Si nanostructures. This approach will advance understanding of cellular responses to extracellular biophysical and biochemical cues. which is likely to facilitate the design of improved cellular manipulation technologies.

A baseline TiAl-containing ALD electrode is established, with properties in line with reported workfunction (WF) materials for scaled RMG nFETs, values below 4.6eV requiring a 25Å layer. Furthermore, a novel ALD metal-compound material, MX, is shown to enable at least 10Å further scaling of the electrode stack due to its superior scavenging power. It can be finely tuned by the film thickness, allowing for a remarkable 20-30meV WF delta per ALD cycle over a minimum 600meV range. The wet etchability of the electrodes makes multi-Vt and dual-WF integration possible. MX does not degrade transfer characteristics and reliability of RMG FinFETs, while the thinner nWF electrode enables reduced gate resistance, as verified down to 20nm metal gate lengths. For the first time, taking advantage of the MX compound scavenging power to control oxygen filaments, we demonstrate an all-ALD HfO 2 -based ReRAM. Forming voltages match those achieved by optimized PVD contacts, while scaling the active electrode thickness by a factor of 4x, down to 5nm. Conformality of the layers enables vertical-ReRAM architectures with reduced line resistance. We conclude the developed electrode can facilitate both logic scaling beyond the 10nm node, and 3D memory technology.

We have demonstrated, for the first time, a combination of outstanding linearity of analog programming with matched PCM pairs, small analog programming noise, an extremely low resistance drift (R-drift) coefficient (0.005, median) and high endurance for a CVD-based confined phase change memory (PCM) with a thin metallic liner. In-depth analysis of linear analog programming is also presented. MNIST simulations using a pair of these confined PCM devices as a synaptic element yield a high test accuracy of 95%.

We demonstrate outstanding resistance-drift (R-drift) mitigation and void elimination as reliability benefits of a thin metallic liner. By tuning the resistivity of the liner, the confined PCM with a metallic liner yields an extremely low R-drift coefficient (∼0.01). We also show for the first time that confined PCM could have a self-recovering property by incorporating a metallic liner. The experimental results with real-time in-situ transmission electron microscope (TEM) exhibit the robustness of the confined PCM that can recover by itself without any extra circuits.

Smart ICT (Information and Communication Technology) such as “Big Data”, “Cloud computing” and Smart Functionalities such as Stand-alone Self-activating MEMS/Sensors construct Smart Systems which enable IoT (Internet of Things), IoE (Internet of Everything) thus Smart Society. To realize above-mentioned Smart Technologies, high-density, low-power consumption, wide-bandwidth, fast-operation semiconductor devices as well as smart functional devices enabled by integrating functionalities with advanced semiconductor technologies including CMOS technologies are necessary. High-density Packaging technologies such as 3D, 2.5D packaging scheme basing on TSV (through-Si via) technology and 2.1D PWB packaging are among key technologies to satisfy the requirements from the both smart semiconductor devices and smart functional devices. Meanwhile MEMS/Sensors are required as muti-functionalities of stand-alone smart devices for wearable devices including smart phone, an important part of Smart Systems. ULVAC has been continuously developing manufacturing solutions for Smart Technologies. In this talk, our high-density packaging technologies including scallop-free, low-temperature processed TSV solution for via-last packaging scheme and 2.1D packaging solutions on large panel build-up PWB will be introduced.

2018 International Conference on Solid State Devices and Materials,In-situ Plasma Conditioning of InGaAs / High-κ Interface Layers for Defect Density Control Compatible with Scalable FinFET Integration

We will report our development results of phase-change and ferroelectric thin film processing technologies including sputtering, MOCVD and plasma etching as well as manufacturing processes for PCRAM, FRAM and MEMS/Sensor device applications. Thin-film functional material such as phase-change materials and ferroelectric materials have been utilized to form advanced semiconductor and electronic devices for internet of things (IoT) solutions. We are confident our manufacturing technologies for these materials and devices will contribute to realizing next generation Smart Society.

All-solid-state thin-film secondary batteries have come to be recognized as one of the key enabling technologies for stand-alone MEMS/sensor devices which are essential for internet of things. However, metallic lithium, which is commonly used as an anode layer in thin-film secondary batteries, has low melting point, so it is not applicable for high-density packaging including high-temperature process such as through-silicon via and solder-reflow process. Silicon is a promising candidate to replace metallic lithium due to high theoretical capacity. In this work, a-Si/LiPON/LiCoO2 cell which is prepared by sputtering technique confirms good reliability during 150 cycles without cracks, and also presents high-temperature tolerance.