High-bandwidth low-current measurement system for automated and scalable probing of tunnel junctions in liquids.

Tunnel junctions have long been used to immobilize and study the electronic transport properties of single molecules. The sensitivity of tunneling currents to entities in the tunneling gap has generated interest in developing electronic biosensors with single molecule resolution. Tunnel junctions can, for example, be used for sensing bound or unbound DNA, RNA, amino acids, and proteins in liquids. However, manufacturing technologies for on-chip integrated arrays of tunnel junction sensors are still in their infancy, and scalable measurement strategies that allow the measurement of large numbers of tunneling junctions are required to facilitate progress. Here, we describe an experimental setup to perform scalable, high-bandwidth (>10 kHz) measurements of low currents (pA-nA) in arrays of on-chip integrated tunnel junctions immersed in various liquid media. Leveraging a commercially available compact 100 kHz bandwidth low-current measurement instrument, we developed a custom two-terminal probe on which the amplifier is directly mounted to decrease parasitic probe capacitances to sub-pF levels. We also integrated a motorized three-axis stage, which could be powered down using software control, inside the Faraday cage of the setup. This enabled automated data acquisition on arrays of tunnel junctions without worsening the noise floor despite being inside the Faraday cage. A deliberately positioned air gap in the fluidic path ensured liquid perfusion to the chip from outside the Faraday cage without coupling in additional noise. We demonstrate the performance of our setup using rapid current switching observed in electromigrated gold tunnel junctions immersed in deionized water.

Electromigrated Gold Nanogap Tunnel Junction Arrays: Fabrication and Electrical Behavior in Liquid and Gaseous Media.

Tunnel junctions have been suggested as high-throughput electronic single molecule sensors in liquids with several seminal experiments conducted using break junctions with reconfigurable gaps. For practical single molecule sensing applications, arrays of on-chip integrated fixed-gap tunnel junctions that can be built into compact systems are preferable. Fabricating nanogaps by electromigration is one of the most promising approaches to realize on-chip integrated tunnel junction sensors. However, the electrical behavior of fixed-gap tunnel junctions immersed in liquid media has not been systematically studied to date, and the formation of electromigrated nanogap tunnel junctions in liquid media has not yet been demonstrated. In this work, we perform a comparative study of the formation and electrical behavior of arrays of gold nanogap tunnel junctions made by feedback-controlled electromigration immersed in various liquid and gaseous media (deionized water, mesitylene, ethanol, nitrogen, and air). We demonstrate that tunnel junctions can be obtained from microfabricated gold nanoconstrictions inside liquid media. Electromigration of junctions in air produces the highest yield (61-67%), electromigration in deionized water and mesitylene results in a lower yield than in air (44-48%), whereas electromigration in ethanol fails to produce viable tunnel junctions due to interfering electrochemical processes. We map out the stability of the conductance characteristics of the resulting tunnel junctions and identify medium-specific operational conditions that have an impact on the yield of forming stable junctions. Furthermore, we highlight the unique challenges associated with working with arrays of large numbers of tunnel junctions in batches. Our findings will inform future efforts to build single molecule sensors using on-chip integrated tunnel junctions.

3D Printing of Glass Micro-Optics with Subwavelength Features on Optical Fiber Tips.

Integration of functional materials and structures on the tips of optical fibers has enabled various applications in micro-optics, such as sensing, imaging, and optical trapping. Direct laser writing is a 3D printing technology that holds promise for fabricating advanced micro-optical structures on fiber tips. To date, material selection has been limited to organic polymer-based photoresists because existing methods for 3D direct laser writing of inorganic materials involve high-temperature processing that is not compatible with optical fibers. However, organic polymers do not feature stability and transparency comparable to those of inorganic glasses. Herein, we demonstrate 3D direct laser writing of inorganic glass with a subwavelength resolution on optical fiber tips. We show two distinct printing modes that enable the printing of solid silica glass structures ("Uniform Mode") and self-organized subwavelength gratings ("Nanograting Mode"), respectively. We illustrate the utility of our approach by printing two functional devices: (1) a refractive index sensor that can measure the indices of binary mixtures of acetone and methanol at near-infrared wavelengths and (2) a compact polarization beam splitter for polarization control and beam steering in an all-in-fiber system. By combining the superior material properties of glass with the plug-and-play nature of optical fibers, this approach enables promising applications in fields such as fiber sensing, optical microelectromechanical systems (MEMS), and quantum photonics.

Cleanroom-Free Direct Laser Micropatterning of Polymers for Organic Electrochemical Transistors in Logic Circuits and Glucose Biosensors.

Organic electrochemical transistors (OECTs) are promising devices for bioelectronics, such as biosensors. However, current cleanroom-based microfabrication of OECTs hinders fast prototyping and widespread adoption of this technology for low-volume, low-cost applications. To address this limitation, a versatile and scalable approach for ultrafast laser microfabrication of OECTs is herein reported, where a femtosecond laser to pattern insulating polymers (such as parylene C or polyimide) is first used, exposing the underlying metal electrodes serving as transistor terminals (source, drain, or gate). After the first patterning step, conducting polymers, such as poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS), or semiconducting polymers, are spin-coated on the device surface. Another femtosecond laser patterning step subsequently defines the active polymer area contributing to the OECT performance by disconnecting the channel and gate from the surrounding spin-coated film. The effective OECT width can be defined with high resolution (down to 2 µm) in less than a second of exposure. Micropatterning the OECT channel area significantly improved the transistor switching performance in the case of PEDOT:PSS-based transistors, speeding up the devices by two orders of magnitude. The utility of this OECT manufacturing approach is demonstrated by fabricating complementary logic (inverters) and glucose biosensors, thereby showing its potential to accelerate OECT research.

Probabilistic cell seeding and non-autofluorescent 3D-printed structures as scalable approach for multi-level co-culture modeling.

To model complex biological tissue in vitro, a specific layout for the position and numbers of each cell type is necessary. Establishing such a layout requires manual cell placement in three dimensions (3D) with micrometric precision, which is complicated and time-consuming. Moreover, 3D printed materials used in compartmentalized microfluidic models are opaque or autofluorescent, hindering parallel optical readout and forcing serial characterization methods, such as patch-clamp probing. To address these limitations, we introduce a multi-level co-culture model realized using a parallel cell seeding strategy of human neurons and astrocytes on 3D structures printed with a commercially available non-autofluorescent resin at micrometer resolution. Using a two-step strategy based on probabilistic cell seeding, we demonstrate a human neuronal monoculture that forms networks on the 3D printed structure and can establish cell-projection contacts with an astrocytic-neuronal co-culture seeded on the glass substrate. The transparent and non-autofluorescent printed platform allows fluorescence-based immunocytochemistry and calcium imaging. This approach provides facile multi-level compartmentalization of different cell types and routes for pre-designed cell projection contacts, instrumental in studying complex tissue, such as the human brain.

Three-dimensional printing of silica glass with sub-micrometer resolution.

Silica glass is a high-performance material used in many applications such as lenses, glassware, and fibers. However, modern additive manufacturing of micro-scale silica glass structures requires sintering of 3D-printed silica-nanoparticle-loaded composites at ~1200 °C, which causes substantial structural shrinkage and limits the choice of substrate materials. Here, 3D printing of solid silica glass with sub-micrometer resolution is demonstrated without the need of a sintering step. This is achieved by locally crosslinking hydrogen silsesquioxane to silica glass using nonlinear absorption of sub-picosecond laser pulses. The as-printed glass is optically transparent but shows a high ratio of 4-membered silicon-oxygen rings and photoluminescence. Optional annealing at 900 °C makes the glass indistinguishable from fused silica. The utility of the approach is demonstrated by 3D printing an optical microtoroid resonator, a luminescence source, and a suspended plate on an optical-fiber tip. This approach enables promising applications in fields such as photonics, medicine, and quantum-optics.

Ultrafast and Resist-Free Nanopatterning of 2D Materials by Femtosecond Laser Irradiation.

The performance of two-dimensional (2D) materials is promising for electronic, photonic, and sensing devices since they possess large surface-to-volume ratios, high mechanical strength, and broadband light sensitivity. While significant advances have been made in synthesizing and transferring 2D materials onto different substrates, there is still the need for scalable patterning of 2D materials with nanoscale precision. Conventional lithography methods require protective layers such as resist or metals that can contaminate or degrade the 2D materials and deteriorate the final device performance. Current resist-free patterning methods are limited in throughput and typically require custom-made equipment. To address these limitations, we demonstrate the noncontact and resist-free patterning of platinum diselenide (PtSe2), molybdenum disulfide (MoS2), and graphene layers with nanoscale precision at high processing speed while preserving the integrity of the surrounding material. We use a commercial, off-the-shelf two-photon 3D printer to directly write patterns in the 2D materials with features down to 100 nm at a maximum writing speed of 50 mm/s. We successfully remove a continuous film of 2D material from a 200 µm × 200 µm substrate area in less than 3 s. Since two-photon 3D printers are becoming increasingly available in research laboratories and industrial facilities, we expect this method to enable fast prototyping of devices based on 2D materials across various research areas.

Microneedle Patch for Painless Intradermal Collection of Interstitial Fluid Enabling Multianalyte Measurement of Small Molecules, SARS-CoV-2 Antibodies, and Protein Profiling.

Blood sampling is a common practice to monitor health, but it entails a series of drawbacks for patients including pain and discomfort. Thus, there is a demand for more convenient ways to obtain samples. Modern analytical techniques enable monitoring of multiple bioanalytes in smaller samples, opening possibilities for new matrices, and microsampling technologies to be adopted. Interstitial fluid (ISF) is an attractive alternative matrix that shows good correlation with plasma concentration dynamics for several analytes and can be sampled in a minimally invasive and painless manner from the skin at the point-of-care. However, there is currently a lack of sampling devices compatible with clinical translation. Here, to tackle state-of-the-art limitations, a cost-effective and compact single-microneedle-based device designed to painlessly collect precisely 1.1 µL of dermal ISF within minutes is presented. The fluid is volume-metered, dried, and stably stored into analytical-grade paper within the microfluidic device. The obtained sample can be mailed to a laboratory, quantitatively analyzed, and provide molecular insights comparable to blood testing. In a human study, the possibility to monitor various classes of molecular analytes is demonstrated in ISF microsamples, including caffeine, hundreds of proteins, and SARS-CoV-2 antibodies, some being detected in ISF for the first time.

Microfluidic Device for Patient-Centric Multiplexed Assays with Readout in Centralized Laboratories.

Patient-centric sampling strategies, where the patient performs self-sampling and ships the sample to a centralized laboratory for readout, are on the verge of widespread adaptation. However, the key to a successful patient-centric workflow is user-friendliness, with few noncritical user interactions, and simple, ideally biohazard-free shipment. Here, we present a capillary-driven microfluidic device designed to perform the critical biomarker capturing step of a multiplexed immunoassay at the time of sample collection. On-chip sample drying enables biohazard-free shipment and allows us to make use of advanced analytics of specialized laboratories that offer the needed analytical sensitivity, reliability, and affordability. Using C-Reactive Protein, MCP1, S100B, IGFBP1, and IL6 as model blood biomarkers, we demonstrate the multiplexing capability and applicability of the device to a patient-centric workflow. The presented quantification of a biomarker panel opens up new possibilities for e-doctor and e-health applications.

Corrigendum to "Probabilistic cell seeding and non-autofluorescent 3D-printed structures as scalable approach for multi-level co-culture modeling" [Mater. Today Bio 21, August 2023, 100706].

[This corrects the article DOI: 10.1016/j.mtbio.2023.100706.].

Micro 3D printing of a functional MEMS accelerometer.

Microelectromechanical system (MEMS) devices, such as accelerometers, are widely used across industries, including the automotive, consumer electronics, and medical industries. MEMS are efficiently produced at very high volumes using large-scale semiconductor manufacturing techniques. However, these techniques are not viable for the cost-efficient manufacturing of specialized MEMS devices at low- and medium-scale volumes. Thus, applications that require custom-designed MEMS devices for markets with low- and medium-scale volumes of below 5000-10,000 components per year are extremely difficult to address efficiently. The 3D printing of MEMS devices could enable the efficient realization and production of MEMS devices at these low- and medium-scale volumes. However, current micro-3D printing technologies have limited capabilities for printing functional MEMS. Herein, we demonstrate a functional 3D-printed MEMS accelerometer using 3D printing by two-photon polymerization in combination with the deposition of a strain gauge transducer by metal evaporation. We characterized the responsivity, resonance frequency, and stability over time of the MEMS accelerometer. Our results demonstrate that the 3D printing of functional MEMS is a viable approach that could enable the efficient realization of a variety of custom-designed MEMS devices, addressing new application areas that are difficult or impossible to address using conventional MEMS manufacturing.

Self-sealing MEMS spray-nozzles to prevent bacterial contamination of portable inhalers for aqueous drug delivery.

Pulmonary drug delivery by portable inhalers is the gold standard in lung disease therapy. An increasing focus on environmentally friendly inhalation currently spurs the development of propellant-free devices. However, the absence of propellants in the drug creates a need for suitable sealing systems that can ensure the pathogenic safety of devices. Traditionally, liquid drug inhalers incorporate a spray nozzle and a separate check valve. Here we show a fully integrated MEMS-based spray system for aqueous drug solutions and demonstrate its bacterial safety. The device comprises a thin silicon membrane with spray orifices, which self-seal against a compliant parylene valve seat underneath. This sealing system prevents bacterial ingrowth in its default closed state, while actuation lifts the membrane from the valve seat upon pressurization and sprays an inhalable aerosol from the nozzles. To seal against bacterial contamination effectively, we found that a contact force between the valve seat and the membrane (featuring the spray nozzles) is needed. In our testing, both self-sealing and an otherwise identical unvalved version of the spray chip can be bacterially safe in continued use when thoroughly cleaned of excess fluids and subjected to low bacterial loads for brief periods. However, when directly exposed to [Formula: see text] CFU/ml of our test organism Citrobacter rodentium for 24 h, unvalved systems become contaminated in nearly 90% of cases. In contrast, self-sealing spray chips reduced contamination probability by 70%. This development may enable preservative-free drug formulations in portable inhalers that use propellant-free aqueous drug solutions.

Blood cell quantification on dry blood samples: toward patient-centric complete blood counts.

Background: Performing complete blood counts from patients' homes could have a transformative impact on e-based healthcare. Blood microsampling and sample drying are enabling elements for patient-centric healthcare. The aim of this study was to investigate the potential of dry blood samples for image-based cell quantification of red and white blood cells. Methods: A manual sample preparation method was developed and tested for image-based red and white blood cell counting. Results & conclusion: Dry blood samples enable image-based cell counting of red and white blood cells with a good correlation to gold standard hematology analyzer data (average coefficient of variation <6.5%; R2 >0.8) and resolve the basic morphology of white blood cell nuclei. The presented proof-of-principle study is a first step toward patient-centric complete blood counts.

3D Microvascularized Tissue Models by Laser-Based Cavitation Molding of Collagen.

3D tissue models recapitulating human physiology are important for fundamental biomedical research, and they hold promise to become a new tool in drug development. An integrated and defined microvasculature in 3D tissue models is necessary for optimal cell functions. However, conventional bioprinting only allows the fabrication of hydrogel scaffolds containing vessel-like structures with large diameters (>100 µm) and simple geometries. Recent developments in laser photoablation enable the generation of this type of structure with higher resolution and complexity, but the photo-thermal process can compromise cell viability and hydrogel integrity. To address these limitations, the present work reports in situ 3D patterning of collagen hydrogels by femtosecond laser irradiation to create channels and cavities with diameters ranging from 20 to 60 µm. In this process, laser irradiation of the hydrogel generates cavitation gas bubbles that rearrange the collagen fibers, thereby creating stable microchannels. Such 3D channels can be formed in cell- and organoid-laden hydrogel without affecting the viability outside the lumen and can enable the formation of artificial microvasculature by the culture of endothelial cells and cell media perfusion. Thus, this method enables organs-on-a-chip and 3D tissue models featuring complex microvasculature.

A large-area single-filament infrared emitter and its application in a spectroscopic ethanol gas sensing system.

Nondispersive infrared (NDIR) spectroscopy is an important technology for highly accurate and maintenance-free sensing of gases, such as ethanol and carbon dioxide. However, NDIR spectroscopy systems are currently too expensive, e.g., for consumer and automotive applications, as the infrared (IR) emitter is a critical but costly component of these systems. Here, we report on a low-cost large-area IR emitter featuring a broadband emission spectrum suitable for small NDIR gas spectroscopy systems. The infrared emitter utilizes Joule heating of a Kanthal (FeCrAl) filament that is integrated in the base substrate using an automated high-speed wire bonding process, enabling simple and rapid formation of a long meander-shaped filament. We describe the critical infrared emitter characteristics, including the effective infrared emission spectrum, thermal frequency response, and power consumption. Finally, we integrate the emitter into a handheld breath alcohol analyzer and show its operation in both laboratory and real-world settings, thereby demonstrating the potential of the emitter for future low-cost optical gas sensor applications.

Large-area integration of two-dimensional materials and their heterostructures by wafer bonding.

Integrating two-dimensional (2D) materials into semiconductor manufacturing lines is essential to exploit their material properties in a wide range of application areas. However, current approaches are not compatible with high-volume manufacturing on wafer level. Here, we report a generic methodology for large-area integration of 2D materials by adhesive wafer bonding. Our approach avoids manual handling and uses equipment, processes, and materials that are readily available in large-scale semiconductor manufacturing lines. We demonstrate the transfer of CVD graphene from copper foils (100-mm diameter) and molybdenum disulfide (MoS2) from SiO2/Si chips (centimeter-sized) to silicon wafers (100-mm diameter). Furthermore, we stack graphene with CVD hexagonal boron nitride and MoS2 layers to heterostructures, and fabricate encapsulated field-effect graphene devices, with high carrier mobilities of up to [Formula: see text]. Thus, our approach is suited for backend of the line integration of 2D materials on top of integrated circuits, with potential to accelerate progress in electronics, photonics, and sensing.

A microfluidic device for TEM sample preparation.

Transmission electron microscopy (TEM) allows for visualizing and analyzing viral particles and has become a vital tool for the development of vaccines and biopharmaceuticals. However, appropriate TEM sample preparation is typically done manually which introduces operator-based dependencies and can lead to unreliable results. Here, we present a capillary-driven microfluidic single-use device that prepares a TEM grid with minimal and non-critical user interaction. The user only initiates the sample preparation process, waits for about one minute and then collects the TEM grid, ready for imaging. Using Adeno-associated virus (AAV) particles as the sample and NanoVan® as the stain, we demonstrate microfluidic consistency and show that the sample preparation quality is sufficient for automated image analysis. We further demonstrate the versatility of the microfluidic device by preparing two protein complexes for TEM investigations using two different stain types. The presented TEM sample preparation concept could alleviate the problems associated with human inconsistency in manual preparation protocols and allow for non-specialists to prepare TEM samples.

Bactericidal surfaces prepared by femtosecond laser patterning and layer-by-layer polyelectrolyte coating.

Antimicrobial surfaces are important in medical, clinical, and industrial applications, where bacterial infection and biofouling may constitute a serious threat to human health. Conventional approaches against bacteria involve coating the surface with antibiotics, cytotoxic polymers, or metal particles. However, these types of functionalization have a limited lifetime and pose concerns in terms of leaching and degradation of the coating. Thus, there is a great interest in developing long-lasting and non-leaching bactericidal surfaces. To obtain a bactericidal surface, we combine micro and nanoscale patterning of borosilicate glass surfaces by ultrashort pulsed laser irradiation and a non-leaching layer-by-layer polyelectrolyte modification of the surface. The combination of surface structure and surface charge results in an enhanced bactericidal effect against both Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria. The laser patterning and the layer-by-layer modification are environmentally friendly processes that are applicable to a wide variety of materials, which makes this method uniquely suited for fundamental studies of bacteria-surface interactions and paves the way for its applications in a variety of fields, such as in hygiene products and medical devices.

Water in contact with the backside of a silicon substrate enables drilling of high-quality holes through the substrate using ultrashort laser pulses.

Holes through silicon substrates are used in silicon microsystems, for example in vertical electrical interconnects. In comparison to deep reactive ion etching, laser drilling is a versatile method for forming these holes, but laser drilling suffers from poor hole quality. In this article, water is used in the silicon drilling process to remove debris and the shape deformations of the holes. Water is introduced into the drilling process through the backside of the substrate to minimize negative effects to the drilling process. Drilling of inclined holes is also demonstrated. The inclined holes could find applications in radio frequency devices.

Vertical integration of microchips by magnetic assembly and edge wire bonding.

The out-of-plane integration of microfabricated planar microchips into functional three-dimensional (3D) devices is a challenge in various emerging MEMS applications such as advanced biosensors and flow sensors. However, no conventional approach currently provides a versatile solution to vertically assemble sensitive or fragile microchips into a separate receiving substrate and to create electrical connections. In this study, we present a method to realize vertical magnetic-field-assisted assembly of discrete silicon microchips into a target receiving substrate and subsequent electrical contacting of the microchips by edge wire bonding, to create interconnections between the receiving substrate and the vertically oriented microchips. Vertical assembly is achieved by combining carefully designed microchip geometries for shape matching and striped patterns of the ferromagnetic material (nickel) on the backside of the microchips, enabling controlled vertical lifting directionality independently of the microchip's aspect ratio. To form electrical connections between the receiving substrate and a vertically assembled microchip, featuring standard metallic contact electrodes only on its frontside, an edge wire bonding process was developed to realize ball bonds on the top sidewall of the vertically placed microchip. The top sidewall features silicon trenches in correspondence to the frontside electrodes, which induce deformation of the free air balls and result in both mechanical ball bond fixation and around-the-edge metallic connections. The edge wire bonds are realized at room temperature and show minimal contact resistance (<0.2 Ω) and excellent mechanical robustness (>168 mN in pull tests). In our approach, the microchips and the receiving substrate are independently manufactured using standard silicon micromachining processes and materials, with a subsequent heterogeneous integration of the components. Thus, this integration technology potentially enables emerging MEMS applications that require 3D out-of-plane assembly of microchips.