Distinct sub-second dopamine signaling in dorsolateral striatum measured by a genetically-encoded fluorescent sensor.

The development of genetically encoded dopamine sensors such as dLight has provided a new approach to measuring slow and fast dopamine dynamics both in brain slices and in vivo, possibly enabling dopamine measurements in areas like the dorsolateral striatum (DLS) where previously such recordings with fast-scan cyclic voltammetry (FSCV) were difficult. To test this, we first evaluated dLight photometry in mouse brain slices with simultaneous FSCV and found that both techniques yielded comparable results, but notable differences in responses to dopamine transporter inhibitors, including cocaine. We then used in vivo fiber photometry with dLight in mice to examine responses to cocaine in DLS. We also compared dopamine responses during Pavlovian conditioning across the striatum. We show that dopamine increases were readily detectable in DLS and describe transient dopamine kinetics, as well as slowly developing signals during conditioning. Overall, our findings indicate that dLight photometry is well suited to measuring dopamine dynamics in DLS.

A Guide to Fluorescence Lifetime Microscopy and Förster's Resonance Energy Transfer in Neuroscience.

Fluorescence lifetime microscopy (FLIM) and Förster's resonance energy transfer (FRET) are advanced optical tools that neuroscientists can employ to interrogate the structure and function of complex biological systems in vitro and in vivo using light. In neurobiology they are primarily used to study protein-protein interactions, to study conformational changes in protein complexes, and to monitor genetically encoded FRET-based biosensors. These methods are ideally suited to optically monitor changes in neurons that are triggered optogenetically. Utilization of this technique by neuroscientists has been limited, since a broad understanding of FLIM and FRET requires familiarity with the interactions of light and matter on a quantum mechanical level, and because the ultra-fast instrumentation used to measure fluorescent lifetimes and resonance energy transfer are more at home in a physics lab than in a biology lab. In this overview, we aim to help neuroscientists overcome these obstacles and thus feel more comfortable with the FLIM-FRET method. Our goal is to aid researchers in the neuroscience community to achieve a better understanding of the fundamentals of FLIM-FRET and encourage them to fully leverage its powerful ability as a research tool. Published 2020. U.S. Government.

Fabry-Pérot optical sensor and portable detector for monitoring high-resolution ocular hemodynamics.

Our understanding of ocular hemodynamics and its role in ophthalmic disease progression remains unclear due to the shortcomings of precise and on-demand biomedical sensing technologies. Here, we report high-resolution in vivo assessment of ocular hemodynamics using a Fabry-Pérot cavity-based micro-optical sensor and a portable optical detector. The designed optical system is capable of measuring both static intraocular pressure and dynamic ocular pulsation profiles in parallel. Through a dynamic intensity variation analysis method which improves sensing resolution by 3-4 folds, our system is able to extract systolic/diastolic phases from a single ocular pulsation profile. Using a portable detector, we performed in vivo studies on rabbits and verified that ophthalmic parameters obtained from our optical system closely match with traditional techniques such as tonometry, electrocardiography, and photo-plethysmography.

Glucose Sensing Using Surface-Enhanced Raman-Mode Constraining.

Diabetes mellitus is a chronic disease, and its management focuses on monitoring and lowering a patient's glucose level to prevent further complications. By tracking the glucose-induced shift in the surface-enhanced Raman-scattering (SERS) emission of mercaptophenylboronic acid (MPBA), we have demonstrated fast and continuous glucose sensing in the physiologically relevant range from 0.1 to 30 mM and verified the underlying mechanism using numerical simulations. Bonding of glucose to MPBA suppresses the "breathing" mode of MPBA at 1071 cm-1 and energizes the constrained-bending mode at 1084 cm-1, causing the dominant peak to shift from 1071 to 1084 cm-1. MPBA-glucose bonding is also reversible, allowing continuous tracking of ambient glucose concentrations, and the MPBA-coated substrates showed very stable performance over a 30 day period, making the approach promising for long-term continuous glucose monitoring. Using Raman-mode-constrained, miniaturized SERS implants, we also successfully demonstrated intraocular glucose measurements in six ex vivo rabbit eyes within ±0.5 mM of readings obtained using a commercial glucose sensor.

Multifunctional biophotonic nanostructures inspired by the longtail glasswing butterfly for medical devices.

Numerous living organisms possess biophotonic nanostructures that provide colouration and other diverse functions for survival. While such structures have been actively studied and replicated in the laboratory, it remains unclear whether they can be used for biomedical applications. Here, we show a transparent photonic nanostructure inspired by the longtail glasswing butterfly (Chorinea faunus) and demonstrate its use in intraocular pressure (IOP) sensors in vivo. We exploit the phase separation between two immiscible polymers (poly(methyl methacrylate) and polystyrene) to form nanostructured features on top of a Si3N4 substrate. The membrane thus formed shows good angle-independent white-light transmission, strong hydrophilicity and anti-biofouling properties, which prevent adhesion of proteins, bacteria and eukaryotic cells. We then developed a microscale implantable IOP sensor using our photonic membrane as an optomechanical sensing element. Finally, we performed in vivo testing on New Zealand white rabbits, which showed that our device reduces the mean IOP measurement variation compared with conventional rebound tonometry without signs of inflammation.

Effect of optical aberrations on intraocular pressure measurements using a microscale optical implant in ex vivo rabbit eyes.

Elevated intraocular pressure (IOP) is the only modifiable major risk factor of glaucoma. Recently, accurate and continuous IOP monitoring has been demonstrated in vivo using an implantable sensor based on optical resonance with remote optical readout to improve patient outcomes. Here, we investigate the relationship between optical aberrations of ex vivo rabbit eyes and the performance of the IOP sensor using a custom-built setup integrated with a Shack-Hartmann sensor. The sensor readouts became less accurate as the aberrations increased in magnitude, but they remained within the clinically acceptable range. For root-mean-square wavefront errors of 0.10 to 0.94 µm, the accuracy and the signal-to-noise ratio were 0.58 ± 0.32 mm Hg and 15.57 ± 4.85 dB, respectively.

>1000-Fold Lifetime Extension of a Nickel Electromechanical Contact Device via Graphene.

Micro-/nano-electromechanical (M/NEM) switches have received significant attention as promising switching devices for a wide range of applications such as computing, radio frequency communication, and power gating devices. However, M/NEM switches still suffer from unacceptably low reliability because of irreversible degradation at the contacting interfaces, hindering adoption in practical applications and further development. Here, we evaluate and verify graphene as a contact material for reliability-enhanced M/NEM switching devices. Atomic force microscopy experiments and quantum mechanics calculations reveal that energy-efficient mechanical contact-separation characteristics are achieved when a few layers of graphene are used as a contact material on a nickel surface, reducing the energy dissipation by 96.6% relative to that of a bare nickel surface. Importantly, graphene displays almost elastic contact-separation, indicating that little atomic-scale wear, including plastic deformation, fracture, and atomic attrition, is generated. We also develop a feasible fabrication method to demonstrate a MEM switch, which has high-quality graphene as the contact material, and verify that the devices with graphene show mechanically stable and elastic-like contact properties, consistent with our nanoscale contact experiment. The graphene coating extends the switch lifetime >103 times under hot switching conditions.

Nanomechanical Encoding Method Using Enhanced Thermal Concentration on a Metallic Nanobridge.

We present a fast, energy-efficient nano-thermomechanical encoding scheme for digital information storage and retrieval. Digital encoding processes are conducted by the bistable electrothermal actuation of a scalable nanobridge device. The electrothermal energy is highly concentrated by enhanced electron/phonon scattering and heat insulation in a sub-100 nm metallic layer. The efficient conversion of electrothermal energy into mechanical strain allows digital switching and programming processes within 60 ns at 0.75 V with a programming energy of only 54 pJ. Furthermore, this encoding scheme together with the thermally robust design enables data retention at temperatures up to 400 °C. These results suggest that the proposed nano-thermomechanical encoding method could contribute to low-power electronics and robust information storage/retrieval systems.

Biocompatible Multifunctional Black-Silicon for Implantable Intraocular Sensor.

Multifunctional black-silicon (b-Si) integrated on the surface of an implantable intraocular pressure sensor significantly improves sensor performance and reliability in six-month in vivo studies. The antireflective properties of b-Si triples the signal-to-noise ratio and increases the optical readout distance to a clinically viable 12 cm. Tissue growth and inflammation response on the sensor is suppressed demonstrating desirable anti-biofouling properties.

A microscale optical implant for continuous in vivo monitoring of intraocular pressure.

Intraocular pressure (IOP) is a key clinical parameter in glaucoma management. However, despite the potential utility of daily measurements of IOP in the context of disease management, the necessary tools are currently lacking, and IOP is typically measured only a few times a year. Here we report on a microscale implantable sensor that could provide convenient, accurate, on-demand IOP monitoring in the home environment. When excited by broadband near-infrared (NIR) light from a tungsten bulb, the sensor's optical cavity reflects a pressure-dependent resonance signature that can be converted to IOP. NIR light is minimally absorbed by tissue and is not perceived visually. The sensor's nanodot-enhanced cavity allows for a 3-5 cm readout distance with an average accuracy of 0.29 mm Hg over the range of 0-40 mm Hg. Sensors were mounted onto intraocular lenses or silicone haptics and secured inside the anterior chamber in New Zealand white rabbits. Implanted sensors provided continuous in vivo tracking of short-term transient IOP elevations and provided continuous measurements of IOP for up to 4.5 months.

Real-Time In Vivo Intraocular Pressure Monitoring using an Optomechanical Implant and an Artificial Neural Network.

Optimized glaucoma therapy requires frequent monitoring and timely lowering of elevated intraocular pressure (IOP). A recently developed microscale IOP-monitoring implant, when illuminated with broadband light, reflects a pressure-dependent optical spectrum that is captured and converted to measure IOP. However, its accuracy is limited by background noise and the difficulty of modeling non-linear shifts of the spectra with respect to pressure changes. Using an end-to-end calibration system to train an artificial neural network (ANN) for signal demodulation we improved the speed and accuracy of pressure measurements obtained with an optically probed IOP-monitoring implant and make it suitable for real-time in vivo IOP monitoring. The ANN converts captured optical spectra into corresponding IOP levels. We achieved an IOP-measurement accuracy of ±0.1 mmHg at a measurement rate of 100 Hz, which represents a ten-fold improvement from previously reported values. This technique allowed real-time tracking of artificially induced sub-1 s transient IOP elevations and minor fluctuations induced by the respiratory motion of the rabbits during in vivo monitoring. All in vivo sensor readings paralleled those obtained concurrently using a commercial tonometer and showed consistency within ±2 mmHg. Real-time processing is highly useful for IOP monitoring in clinical settings and home environments and improves the overall practicality of the optical IOP-monitoring approach.

High-performance hybrid complementary logic inverter through monolithic integration of a MEMS switch and an oxide TFT.

A hybrid complementary logic inverter consisting of a microelectromechanical system switch as a promising alternative for the p-type oxide thin film transistor (TFT) and an n-type oxide TFT is presented for ultralow power integrated circuits. These heterogeneous microdevices are monolithically integrated. The resulting logic device shows a distinctive voltage transfer characteristic curve, very low static leakage, zero-short circuit current, and exceedingly high voltage gain.

High throughput ultralong (20 cm) nanowire fabrication using a wafer-scale nanograting template.

Nanowires are being actively explored as promising nanostructured materials for high performance flexible electronics, biochemical sensors, photonic applications, solar cells, and secondary batteries. In particular, ultralong (centimeter-long) nanowires are highly attractive from the perspective of electronic performance, device throughput (or productivity), and the possibility of novel applications. However, most previous works on ultralong nanowires have issues related to limited length, productivity, difficult alignment, and deploying onto the planar substrate complying with well-matured device fabrication technologies. Here, we demonstrate a highly ordered ultralong (up to 20 cm) nanowire array, with a diameter of 50 nm (aspect ratio of up to 4,000,000:1), in an unprecedented large (8 in.) scale (2,000,000 strands on a wafer). We first devised a perfectly connected ultralong nanograting master template on the whole area of an 8 in. substrate using a top-down approach, with a density equivalent to that achieved with e-beam lithography (100 nm). Using this large-area, ultralong, high-density nanograting template, we developed a fast and effective method for fabricating up to 20 cm long nanowire arrays on a plastic substrate, composed of metal, dielectric, oxide, and ferroelectric materials. As a suggestion of practical application, a prototype of a large-area aluminum wire grid polarizer was demonstrated.

A sub-1-volt nanoelectromechanical switching device.

Nanoelectromechanical (NEM) switches have received widespread attention as promising candidates in the drive to surmount the physical limitations currently faced by complementary metal oxide semiconductor technology. The NEM switch has demonstrated superior characteristics including quasi-zero leakage behaviour, excellent density capability and operation in harsh environments. However, an unacceptably high operating voltage (4-20 V) has posed a major obstacle in the practical use of the NEM switch in low-power integrated circuits. To utilize the NEM switch widely as a core device component in ultralow power applications, the operation voltage needs to be reduced to 1 V or below. However, sub-1 V actuation has not yet been demonstrated because of fabrication difficulties and irreversible switching failure caused by surface adhesion. Here, we report the sub-1 V operation of a NEM switch through the introduction of a novel pipe clip device structure and an effective air gap fabrication technique. This achievement is primarily attributed to the incorporation of a 4-nm-thick air gap, which is the smallest reported so far for a NEM switch generated using a 'top-down' approach. Our structure and process can potentially be utilized in various nanogap-related applications, including NEM switch-based ultralow-power integrated circuits, NEM resonators, nanogap electrodes for scientific research and sensors.

A CMOS label-free DNA sensor using electrostatic induction of molecular charges.

This paper reports a label-free biosensor for the detection of DNA hybridization. The proposed biosensor measures the surface potential on oligonucleotide modified electrodes using a direct charge accumulation method. The sensor directly and repeatedly measures the charges induced in the working electrode, which correspond to intrinsic negative charges in immobilized molecules. The sensor achieves an improved signal-to-noise ratio (SNR), through the oversampling effect of accumulation for charges and the differential architecture. The sensor also shows stable, robust, and reproducible measurement independent of slight changes in the reference voltage, unlike previous ion-sensitive field effect transistors (ISFETs), providing the benefits of choosing a wide variety of reference electrode materials. The proposed device is integrated with working electrodes, a reference electrode and readout circuits into one package via a 0.35 µm complementary metal-oxide-semiconductor (CMOS) process. The sensor achieves a detectable range of 88.3 dB and a detection limit of 36 µV for surface potential. It is demonstrated that the sensor successfully achieves specific detection of oligonucleotide sequences derived from the H5N1 avian influenza virus. The experiments show a limit of detection of 100 pM and include a single-base mismatch test in 18-mer oligonucleotides.

An autonomous CMOS hysteretic sensor for the detection of desorption-free DNA hybridization.

This paper describes a sensor for label-free, fully electrical detection of DNA hybridization based on capacitive changes in the electrode-electrolyte interface. The sensor measures capacitive changes in real time according to a charging-discharging principle that is limited by the hysteresis window. In addition, a novel autonomous searching technique, which exclusively monitors desorption-free hybridized electrodes among electrode arrays, enhances the performance of the sensor compared with conventional capacitive measurement. The sensor system achieves a detection range of 80 dB. The integrated circuit sensor is fabricated with a 0.35 µm CMOS process. The proposed sensor offers rapid, robust and inexpensive measurement of capacitance with highly integrated detection circuitry. It also facilitates quantitative evaluations of molecular densities on a chip with distinctive impedance variations by monitoring desorption-free hybridized electrodes. Our electrical biosensor has great potential for use with bio analytical tools and point-of-care diagnosis.

An electronic DNA sensor chip using integrated capacitive read-out circuit.

This paper presents fully integrated label-free DNA recognition circuit based on capacitance measurement. A CMOS-based DNA sensor is implemented for the electrical detection of DNA hybridization. The proposed architecture detects the difference of capacitance through the integration of current mismatch of capacitance between reference electrodes functionalized with only single-stranded DNA and sensing electrodes bound with complementary DNA strands specifically. In addition, to minimize the effects of parallel resistance between electrodes and DNA layers, the compensation technique of leakage current through the use of constant current charging and discharging is implemented in the proposed detection circuit. The chip was fabricated in 0.35um 4-metal 2-poly CMOS process, and 16 × 8 sensing electrode arrays were fabricated by post-processing steps.

One-chip electronic detection of DNA hybridization using precision impedance-based CMOS array sensor.

This paper describes a label-free and fully electronic detection method of DNA hybridization, which is achieved through the use of a 16×8 microarray sensor in conjunction with a new type of impedance spectroscopy constructed with standard complementary metal-oxide-semiconductor (CMOS) technology. The impedance-based method is based on changes in the reactive capacitance and the charge-transfer resistance after hybridization with complementary DNA targets. In previously published label-free techniques, the measured capacitance presented unstable capacitive properties due to the parallel resistance that is not infinite and can cause a leakage by discharging the charge on the capacitor. This paper presents an impedance extraction method that uses excitation by triangular wave voltage, which enables a reliable measurement of both C and R producing a highly sensitive sensor with a stable operation independent of external variables. The system was fabricated in an industrial 0.35-µm 4-metal 2-poly CMOS process, integrating working electrodes and readout electronics into one chip. The integrated readout, which uses a parasitic insensitive integrator, achieves an enlarged detection range and improved noise performance. The maximum average relative variations of C and R are 31.5% and 68.6%, respectively, after hybridization with a 1 µM target DNA. The proposed sensor allows quantitative evaluation of the molecule densities on the chip with distinguishable variation in the impedance. This fully electronic microsystem has great potential for use with bioanalytical tools and point-of-care diagnosis.