Remote Estimation of Blood Pressure Using Millimeter-Wave Frequency-Modulated Continuous-Wave Radar.

This paper proposes to remotely estimate a human subject's blood pressure using a millimeter-wave radar system. High blood pressure is a critical health threat that can lead to diseases including heart attacks, strokes, kidney disease, and vision loss. The commonest method of measuring blood pressure is based on a cuff that is contact-based, non-continuous, and cumbersome to wear. Continuous remote monitoring of blood pressure can facilitate early detection and treatment of heart disease. This paper investigates the possibility of using millimeter-wave frequency-modulated continuous-wave radar to measure the heart blood pressure by means of pulse wave velocity (PWV). PWV is known to be highly correlated with blood pressure, which can be measured by pulse transit time. We measured PWV using a two-millimeter wave radar focused on the subject's chest and wrist. The measured time delay provided the PWV given the length from the chest to the wrist. In addition, we analyzed the measured radar signal from the wrist because the shape of the pulse wave purveyed information on blood pressure. We investigated the area under the curve (AUC) as a feature and found that AUC is strongly correlated with blood pressure. In the experiment, five human subjects were measured 50 times each after performing different activities intended to influence blood pressure. We used artificial neural networks to estimate systolic blood pressure (SBP) and diastolic blood pressure (SBP) with both PWV and AUC as inputs. The resulting root mean square errors of estimated blood pressure were 3.33 mmHg for SBP and 3.14 mmHg for DBP.

Recent Advancements in Technologies to Detect Enterohaemorrhagic Escherichia coli Shiga Toxins.

Shiga toxin (Stxs)-producing enterohaemorrhagic Escherichia coli (EHEC) and Shigella dysenteriae serotype 1 are major causative agents of severe bloody diarrhea (known as hemorrhagic colitis) and hemolytic uremic syndrome (HUS) associated with extraintestinal complications such as acute renal failure and neurologic impairment in infected patients under 9 years of age. Extreme nephrotoxicity of Stxs in HUS patients is associated with severe outcomes, highlighting the need to develop technologies to detect low levels of the toxin in environmental or food samples. Currently, the conventional polymerase chain reaction (PCR) or immunoassay is the most broadly used assay to detect the toxin. However, these assays are laborious, time-consuming, and costly. More recently, numerous studies have described novel, highly sensitive, and portable methods for detecting Stxs from EHEC. To contextualize newly emerging Stxs detection methods, we briefly explain the basic principles of these methods, including lateral flow assays, optical detection, and electrical detection. We subsequently describe existing and newly emerging rapid detection technologies to identify and measure Stxs.

Development of Real-Time Cuffless Blood Pressure Measurement Systems with ECG Electrodes and a Microphone Using Pulse Transit Time (PTT).

Research has shown that pulse transit time (PTT), which is the time delay between the electrocardiogram (ECG) signal and the signal from a photoplethysmogram (PPG) sensor, can be used to estimate systolic blood pressure (SBP) and diastolic blood pressure (DBP) without the need for a cuff. However, the LED of the PPG sensor requires the precise adjustment of both light intensity and light absorption rates according to the contact status of the light-receiving element. This results in the need for regular calibration. In this study, we propose a cuffless blood pressure monitor that measures real-time blood pressure using a microphone instead of a PPG sensor. The blood pulse wave is measured in the radial artery of the wrist using a microphone that can directly measure the sound generated by a body rather than sending energy inside the body and receiving a returning signal. Our blood pressure monitor uses the PTT between the R-peak of the ECG signal and two feature points of the blood pulse wave in the radial artery of the wrist. ECG electrodes and circuits were fabricated, and a commercial microelectromechanical system (MEMS) microphone was used as the microphone to measure blood pulses. The peak points of the blood pulse from the microphone were clear, so the estimated SBP and DBP could be obtained from each ECG pulse in real time, and the resulting estimations were similar to those made by a commercial cuff blood pressure monitor. Since neither the ECG electrodes nor the microphone requires calibration over time, the real-time cuffless blood pressure monitor does not require calibration. Using the developed device, blood pressure was measured three times daily for five days, and the mean absolute error (MAE) and standard deviation (SD) of the SBP and DBP were found to be 2.72 ± 3.42 mmHg and 2.29 ± 3.53 mmHg, respectively. As a preliminary study for proof-of-concept, these results were obtained from one subject. The next step will be a pilot study on a large number of subjects.

Development and Optimization of Silicon-Dioxide-Coated Capacitive Electrode for Ambulatory ECG Measurement System.

This paper presents a silicon-dioxide-coated capacitive electrode system for an ambulatory electrocardiogram (ECG). The electrode was coated with a nano-leveled (287 nm) silicon dioxide layer which has a very high resistance of over 200 MΩ. Due to this high resistance, the electrode can be defined as only a capacitor without a resistive characteristic. This distinct capacitive characteristic of the electrode brings a simplified circuit analysis to achieve the development of a high-quality ambulatory ECG system. The 240 um thickness electrode was composed of a stainless-steel sheet layer for sensing, a polyimide electrical insulation layer, and a copper sheet connected with the ground to block any electrical noises generated from the back side of the structure. Six different diameter electrodes were prepared to optimize ECG signals in ambulatory environment, such as the amplitude of the QRS complex, amplitude of electromagnetic interference (EMI), and baseline wandering of the ECG signals. By combining the experimental results, optimal ambulatory ECG signals were obtained with electrodes that have a diameter from 1 to 3 cm. Moreover, we achieved high-quality ECG signals in a sweating simulation environment with 2 cm electrodes.

A 3D Miniaturized Glass Magnetic-Active Centrifugal Micropump Fabricated by SLE Process and Laser Welding.

A miniaturized pump to manipulate liquid flow in microchannels is the key component of microfluidic devices. Many researchers have demonstrated active microfluidic pumps, but most of them still required additional large peripherals to operate their micropumps. In addition, those micropumps were made of polymer materials so that their application may be limited to a variety of fields that require harsh conditions at high pressures and temperatures or organic solvents and acid/base. In this work, we present a 3D miniaturized magnetic-driven glass centrifugal pump for microfluidic devices. The pump consists of a volute structure and a 3D impeller integrated with two magnet disks of Φ1 mm. The 3D pump structure was 13 mm × 10.5 mm × 3 mm, and it was monolithically fabricated in a fused silica sheet by selective laser-induced etching (SLE) technology using a femtosecond laser. The pump operation requires only one motor rotating two magnets. It was Φ42 mm × 54 mm and powered by a battery. To align the shaft of the motor to the center of the 3D glass pump chip, a housing containing the motor and the chip was fabricated, and the overall size of the proposed micropump device was 95 mm × 70 mm × 75 mm. Compared with other miniaturized pumps, ours was more compact and portable. The output pressure of the fabricated micropump was between 215 Pa and 3104 Pa, and the volumetric flow rate range was 0.55 mL/min and 7.88 mL/min. The relationship between the motor RPM and the impeller RPM was analyzed, and the flow rate was able to be controlled by the RPM. With its portability, the proposed pump can be applied to produce an integrated and portable microfluidic device for point-of-care analysis.

A Thermocycler Using a Chip Resistor Heater and a Glass Microchip for a Portable and Rapid Microchip-Based PCR Device.

This study proposes a rapid and inexpensive thermocycler that enables rapid heating of samples using a thin glass chip and a cheap chip resistor to overcome the on-site diagnostic limitations of polymerase chain reaction (PCR). Microchip PCR devices have emerged to miniaturize conventional PCR systems and reduce operation time and cost. In general, PCR microchips require a thin-film heater fabricated through a semiconductor process, which is a complicated process, resulting in high costs. Therefore, this investigation substituted a general chip resistor for a thin-film heater. The proposed thermocycler consists of a compact glass microchip of 12.5 mm × 12.5 mm × 2 mm that could hold a 2 µL PCR sample and a surface-mounted chip resistor of 6432 size (6.4 mm × 3.2 mm). Improving heat transfer from the chip resistor heater to the PCR reaction chamber in the microchip was accomplished via the design and fabrication of a three-dimensional chip structure using selective laser-induced etching, a rapid prototyping technique that allowed to be embedded. The fabricated PCR microchip was combined with a thermistor temperature sensor, a blower fan, and a microcontroller. The assembled thermocycler could heat the sample at a maximum rate of 28.8 °C/s per second. When compared with a commercially available PCR apparatus running the same PCR protocol, the total PCR operating time with a DNA sample was reduced by about 20%.

Monolithic 3D micromixer with an impeller for glass microfluidic systems.

The performance of micromixers, namely their mixing efficiency and throughput, is a critical component in increasing the overall efficiency of microfluidic systems (e.g., lab-on-a-chip and µ-TAS). Most previously reported high-performance micromixers use active elements with some external power to induce turbulence, or contain long and complex fluidic channels with obstacles to increase diffusion. In this paper, we introduce a new type of 3D impeller micromixer built within a single fused silica substrate. The proposed device is composed of microchannels with three inlets and a tank, with a mixing impeller passively rotated by axial flow. The passive micromixer is directly fabricated inside a glass plate using a selective laser-induced etching technique. The mixing tank, with its rotating shaft and 3D pitched blade impeller, exists within a micro-cavity with a volume of only 0.28 mm3. A mixing efficiency of 99% is achieved in mixing experiments involving three dye colours over flow rates ranging from 1.5-30 mL min-1, with the same flow rates also applied to a sodium hydroxide-based bromothymol blue indicator and a hydrochloric acid chemical solution. To verify the reliable performance of the proposed device, we compare the mixing index with a general self-circulation-type chamber mixer to demonstrate the improved mixing efficiency achieved by rotating the impeller. No cracking or breakage of the device is observed under high inner pressures or when the maximum flow rate is applied to the mixer. The proposed microfluidic system based on a compact built-in 3D micromixer with an impeller opens the door to robust, highly efficient, and high-throughput glass-based platforms for micro-centrifuges, cell sorters, micro-turbines, and micro-pumps.

A portable and high-sensitivity optical sensing system for detecting fluorescently labeled enterohaemorrhagic Escherichia coli Shiga toxin 2B-subunit.

We developed a stand-alone, real-time optical detection device capable of reading fluorescence intensities from cell samples with high sensitivity and precision, for use as a portable fluorescent sensor for sensing fluorescently labeled enterohemorrhagic Escherichia coli (EHEC) Shiga toxins (Stxs). In general, the signal intensity from the fluorescently labeled Stxs was weak due to the small number of molecules bound to each cell. To address this technical challenge, we used a highly sensitive light detector (photomultiplier tube: PMT) to measure fluorescence, and designed a portable optical housing to align optical parts precisely; the housing itself was fabricated on a 3D printer. In addition, an electric circuit that amplified PMT output was designed and integrated into the system. The system shows the toxin concentration in the sample on a liquid crystal display (LCD), and a microcontroller circuit is used to read PMT output, process data, and display results. In contrast to other portable fluorescent detectors, the system works alone, without any peripheral computer or additional apparatus; its total size is about 17 × 13 × 9 cm3, and it weighs about 770 g. The detection limit was 0.01 ppm of Alexa Fluor 488 in PBS, which is ten thousand times lower than those of other smartphone-based systems and sufficiently sensitive for use with a portable optical detector. We used the portable real-time optical sensing system to detect Alexa Fluor 488-tagged Stx2B-subunits bound to monocytic THP-1 cells expressing the toxin receptor globotriaosylceramide (Gb3). The device did not detect a signal from Gb3-negative PD36 cells, indicating that it was capable of specifically detecting Stxs bound to cells expressing the toxin receptor. Following the development of a rapid and autonomous method for fluorescently tagging cells in food samples, the optical detection system described here could be used for direct detection of Shiga toxins in food in the field.

Characteristics of an Implantable Blood Pressure Sensor Packaged by Ultrafast Laser Microwelding.

We propose a new packaging process for an implantable blood pressure sensor using ultrafast laser micro-welding. The sensor is a membrane type, passive device that uses the change in the capacitance caused by the membrane deformation due to applied pressure. Components of the sensor such as inductors and capacitors were fabricated on two glass (quartz) wafers and the two wafers were bonded into a single package. Conventional bonding methods such as adhesive bonding, thermal bonding, and anodic bonding require considerable effort and cost. Therefore CO2 laser cutting was used due to its fast and easy operation providing melting and bonding of the interface at the same time. However, a severe heat process leading to a large temperature gradient by rapid heating and quenching at the interface causes microcracks in brittle glass and results in low durability and production yield. In this paper, we introduce an ultrafast laser process for glass bonding because it can optimize the heat accumulation inside the glass by a short pulse width within a few picoseconds and a high pulse repetition rate. As a result, the ultrafast laser welding provides microscale bonding for glass pressure sensor packaging. The packaging process was performed with a minimized welding seam width of 100 µm with a minute. The minimized welding seam allows a drastic reduction of the sensor size, which is a significant benefit for implantable sensors. The fabricated pressure sensor was operated with resonance frequencies corresponding to applied pressures and there was no air leakage through the welded interface. In addition, in vitro cytotoxicity tests with the sensor showed that there was no elution of inner components and the ultrafast laser packaged sensor is non-toxic. The ultrafast laser welding provides a fast and robust glass chip packaging, which has advantages in hermeticity, bio-compatibility, and cost-effectiveness in the manufacturing of compact implantable sensors.

Bonding Strength of a Glass Microfluidic Device Fabricated by Femtosecond Laser Micromachining and Direct Welding.

We present a rapid and highly reliable glass (fused silica) microfluidic device fabrication process using various laser processes, including maskless microchannel formation and packaging. Femtosecond laser assisted selective etching was adopted to pattern microfluidic channels on a glass substrate and direct welding was applied for local melting of the glass interface in the vicinity of the microchannels. To pattern channels, a pulse energy of 10 µJ was used with a scanning speed of 100 mm/s at a pulse repetition rate of 500 kHz. After 20⁻30 min of etching in hydrofluoric acid (HF), the glass was welded with a pulse energy of 2.7 µJ and a speed of 20 mm/s. The developed process was as simple as drawing, but powerful enough to reduce the entire production time to an hour. To investigate the welding strength of the fabricated glass device, we increased the hydraulic pressure inside the microchannel of the glass device integrated into a custom-built pressure measurement system and monitored the internal pressure. The glass device showed extremely reliable bonding by enduring internal pressure up to at least 1.4 MPa without any leakage or breakage. The measured pressure is 3.5-fold higher than the maximum internal pressure of the conventional polydimethylsiloxane (PDMS)⁻glass or PDMS⁻PDMS bonding. The demonstrated laser process can be applied to produce a new class of glass devices with reliability in a high pressure environment, which cannot be achieved by PDMS devices or ultraviolet (UV) glued glass devices.

A Microchip for High-throughput Axon Growth Drug Screening.

It has been recently known that not only the presence of inhibitory molecules associated with myelin but also the reduced growth capability of the axons limit mature central nervous system (CNS) axonal regeneration after injury. Conventional axon growth studies are typically conducted using multi-well cell culture plates that are very challenging to investigate localized effects of drugs and limited to low throughput. Unfortunately, there is currently no other in vitro tools that allow investigating localized axonal responses to biomolecules in high-throughput for screening potential drugs that might promote axonal growth. We have developed a compartmentalized neuron culture platform enabling localized biomolecular treatments in parallel to axons that are physically and fluidically isolated from their neuronal somata. The 24 axon compartments in the developed platform are designed to perform four sets of six different localized biomolecular treatments simultaneously on a single device. In addition, the novel microfluidic configuration allows culture medium of 24 axon compartments to be replenished altogether by a single aspiration process, making high-throughput drug screening a reality.

Implantable Bladder Sensors: A Methodological Review.

The loss of urinary bladder control/sensation, also known as urinary incontinence (UI), is a common clinical problem in autistic children, diabetics, and the elderly. UI not only causes discomfort for patients but may also lead to kidney failure, infections, and even death. The increase of bladder urine volume/pressure above normal ranges without sensation of UI patients necessitates the need for bladder sensors. Currently, a catheter-based sensor is introduced directly through the urethra into the bladder to measure pressure variations. Unfortunately, this method is inaccurate because measurement is affected by disturbances in catheter lines as well as delays in response time owing to the inertia of urine inside the bladder. Moreover, this technique can cause infection during prolonged use; hence, it is only suitable for short-term measurement. Development of discrete wireless implantable sensors to measure bladder volume/pressure would allow for long-term monitoring within the bladder, while maintaining the patient's quality of life. With the recent advances in microfabrication, the size of implantable bladder sensors has been significantly reduced. However, microfabricated sensors face hostility from the bladder environment and require surgical intervention for implantation inside the bladder. Here, we explore the various types of implantable bladder sensors and current efforts to solve issues like hermeticity, biocompatibility, drift, telemetry, power, and compatibility issues with popular imaging tools such as computed tomography and magnetic resonance imaging. We also discuss some possible improvements/emerging trends in the design of an implantable bladder sensor.

Microfluidic acoustophoretic force based low-concentration oil separation and detection from the environment.

Detecting and quantifying extremely low concentrations of oil from the environment have broad applications in oil spill monitoring in ocean and coastal areas as well as in oil leakage monitoring on land. Currently available methods for low-concentration oil detection are bulky or costly with limited sensitivities. Thus they are difficult to be used as portable and field-deployable detectors in the case of oil spills or for monitoring the long-term effects of dispersed oil on marine and coastal ecosystems. Here, we present a low-concentration oil droplet trapping and detection microfluidic system based on the acoustophoresis phenomenon where oil droplets in water having a negative acoustic contrast factor move towards acoustic pressure anti-nodes. By trapping oil droplets from water samples flowing through a microfluidic channel, even very low concentrations of oil droplets can be concentrated to a detectable level for further analyses, which is a significant improvement over currently available oil detection systems. Oil droplets in water were successfully trapped and accumulated in a circular acoustophoretic trapping chamber of the microfluidic device and detected using a custom-built compact fluorescent detector based on the natural fluorescence of the trapped crude oil droplets. After the on-line detection, crude oil droplets released from the trapping chamber were successfully separated into a collection outlet by acoustophoretic force for further off-chip analyses. The developed microfluidic system provides a new way of trapping, detecting, and separating low-concentration crude oil from environmental water samples and holds promise as a low-cost field-deployable oil detector with extremely high sensitivity. The microfluidic system and operation principle are expected to be utilized in a wide range of applications where separating, concentrating, and detecting small particles having a negative acoustic contrast factor are required.

Development of a real-time microchip PCR system for portable plant disease diagnosis.

Rapid and accurate detection of plant pathogens in the field is crucial to prevent the proliferation of infected crops. Polymerase chain reaction (PCR) process is the most reliable and accepted method for plant pathogen diagnosis, however current conventional PCR machines are not portable and require additional post-processing steps to detect the amplified DNA (amplicon) of pathogens. Real-time PCR can directly quantify the amplicon during the DNA amplification without the need for post processing, thus more suitable for field operations, however still takes time and require large instruments that are costly and not portable. Microchip PCR systems have emerged in the past decade to miniaturize conventional PCR systems and to reduce operation time and cost. Real-time microchip PCR systems have also emerged, but unfortunately all reported portable real-time microchip PCR systems require various auxiliary instruments. Here we present a stand-alone real-time microchip PCR system composed of a PCR reaction chamber microchip with integrated thin-film heater, a compact fluorescence detector to detect amplified DNA, a microcontroller to control the entire thermocycling operation with data acquisition capability, and a battery. The entire system is 25 × 16 × 8 cm(3) in size and 843 g in weight. The disposable microchip requires only 8-µl sample volume and a single PCR run consumes 110 mAh of power. A DNA extraction protocol, notably without the use of liquid nitrogen, chemicals, and other large lab equipment, was developed for field operations. The developed real-time microchip PCR system and the DNA extraction protocol were used to successfully detect six different fungal and bacterial plant pathogens with 100% success rate to a detection limit of 5 ng/8 µl sample.

Ratiometric temperature imaging using environment-insensitive luminescence of Mn-doped core-shell nanocrystals.

We report a ratiometric temperature imaging method based on Mn luminescence from Mn-doped CdS-ZnS nanocrystals (NCs) with controlled doping location, which is designed to exhibit strong temperature dependence of the spectral lineshape while being insensitive to the surrounding chemical environment. Ratiometric thermometry on the Mn luminescence spectrum was performed by using Mn-doped CdS-ZnS core-shell NCs that have a large local lattice strain on the Mn site, which results in the enhanced temperature dependence of the bandwidth and peak position. The Mn luminescence spectral lineshape is highly robust with respect to the change in the polarity, phase and pH of the surrounding medium and aggregation of the NCs, showing great potential in temperature imaging under chemically heterogeneous environment. The temperature sensitivity (ΔIR/IR = 0.5%/K at 293 K, IR = intensity ratio at two different wavelengths) is highly linear in a wide range of temperatures from cryogenic to above-ambient temperatures. We demonstrate the surface temperature imaging of a cryo-cooling device showing a temperature variation of >200 K by imaging the luminescence of the NC film formed by simple spin coating, taking advantage of the environment-insensitive luminescence.

Microfluidic electro-sonoporation: a multi-modal cell poration methodology through simultaneous application of electric field and ultrasonic wave.

A microfluidic device that simultaneously applies the conditions required for microelectroporation and microsonoporation in a flow-through scheme toward high-efficiency and high-throughput molecular delivery into mammalian cells is presented. This multi-modal poration microdevice using simultaneous application of electric field and ultrasonic wave was realized by a three-dimensional (3D) microelectrode scheme where the electrodes function as both electroporation electrodes and cell flow channel so that acoustic wave can be applied perpendicular to the electric field simultaneously to cells flowing through the microfluidic channel. This 3D microelectrode configuration also allows a uniform electric field to be applied while making the device compatible with fluorescent microscopy. It is hypothesized that the simultaneous application of two different fields (electric field and acoustic wave) in perpendicular directions allows formation of transient pores along two axes of the cell membrane at reduced poration intensities, hence maximizing the delivery efficiency while minimizing cell death. The microfluidic electro-sonoporation system was characterized by delivering small molecules into mammalian cells, and showed average poration efficiency of 95.6% and cell viability of 97.3%. This proof of concept result shows that by combining electroporation and sonoporation together, significant improvement in molecule delivery efficiency could be achieved while maintaining high cell viability compared to electroporation or sonoporation alone. The microfluidic electro-sonoporation device presented here is, to the best of our knowledge, the first multi-modal cell poration device using simultaneous application of electric field and ultrasonic wave. This new multi-modal cell poration strategy and system is expected to have broad applications in delivery of small molecule therapeutics and ultimately in large molecule delivery such as gene transfection applications where high delivery efficiency and high viability are crucial.

A magnetic resonance (MR) microscopy system using a microfluidically cryo-cooled planar coil.

We present the development of a microfluidically cryo-cooled planar coil for magnetic resonance (MR) microscopy. Cryogenically cooling radiofrequency (RF) coils for magnetic resonance imaging (MRI) can improve the signal to noise ratio (SNR) of the experiment. Conventional cryostats typically use a vacuum gap to keep samples to be imaged, especially biological samples, at or near room temperature during cryo-cooling. This limits how close a cryo-cooled coil can be placed to the sample. At the same time, a small coil-to-sample distance significantly improves the MR imaging capability due to the limited imaging depth of planar MR microcoils. These two conflicting requirements pose challenges to the use of cryo-cooling in MR microcoils. The use of a microfluidic based cryostat for localized cryo-cooling of MR microcoils is a step towards eliminating these constraints. The system presented here consists of planar receive-only coils with integrated cryo-cooling microfluidic channels underneath, and an imaging surface on top of the planar coils separated by a thin nitrogen gas gap. Polymer microfluidic channel structures fabricated through soft lithography processes were used to flow liquid nitrogen under the coils in order to cryo-cool the planar coils to liquid nitrogen temperature (-196 °C). Two unique features of the cryo-cooling system minimize the distance between the coil and the sample: (1) the small dimension of the polymer microfluidic channel enables localized cooling of the planar coils, while minimizing thermal effects on the nearby imaging surface. (2) The imaging surface is separated from the cryo-cooled planar coil by a thin gap through which nitrogen gas flows to thermally insulate the imaging surface, keeping it above 0 °C and preventing potential damage to biological samples. The localized cooling effect was validated by simulations, bench testing, and MR imaging experiments. Using this cryo-cooled planar coil system inside a 4.7 Tesla MR system resulted in an average image SNR enhancement of 1.47 ± 0.11 times relative to similar room-temperature coils.