MNBS LAB

시가 독소(Stx)는 인간에게 심각한 위장 질환을 일으키는 것으로 알려진 가장 강력한 박테리아 독소 중 하나입니다. 그러나 시간, 장비 및 운영자 측면에서 높은 비용으로 인해 시기적절한 진단에 추가 적용이 방해됩니다. 본 연구에서는 Stx 수용체인 글로보트리아오실세라미드(Gb3)를 유리 칩에 코팅하여 실시간으로 식품 독소를 검출하는 방법과 칩 장착 후 Stx를 진단할 수 있는 휴대용 형광 검출 시스템을 제공합니다. 유기발광다이오드(OLED) 디스플레이를 탑재한 시스템 전체 크기는 약 8.8 × 6.2 × 15cm3, 무게는 리튬이온 배터리 포함 약 375.5g이다. 새로운 용매 보조 지질 이중층 방법은 이중 지질 막을 배치하고 수용체를 막에 삽입하는 대신 유리 기판에 수용체 이중층을 코팅하여 칩 내의 형광 강도를 높이고 시스템에 의해 감지되는 데 활용되었습니다. 코팅 수용체의 최적 조건이 검증되었으며, 용매 교환 과정에서 유속 3μL/min, 유속 10분에서 비교적 좋은 효율을 보였다. 검출 한계는 250pg/mL의 Stx1B였으며 시스템은 상추 용출액 샘플에서 Stx를 성공적으로 검출했습니다.

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.

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.

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 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.

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%.

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 mu-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 mm(3). 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.

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 enterohemorrhagicEscherichia 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 x 13 x 9 cm(3), 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.

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 m 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.

We present the selective laser-induced etching (SLE) process and design guidelines for the fabrication of three-dimensional (3D) microfluidic channels in a glass. The SLE process consisting of laser direct patterning and wet chemical etching uses different etch rates between the laser modified area and the unmodified area. The etch selectivity is an important factor for the processing speed and the fabrication resolution of the 3D structures. In order to obtain the maximum etching selectivity, we investigated the process window of the SLE process: the laser pulse energy, pulse repetition rate, and scan speed. When using potassium hydroxide (KOH) as a wet etchant, the maximum etch rate of the laser-modified glass was obtained to be 166 μm/h, exhibiting the highest selectivity about 333 respect to the pristine glass. Based on the optimized process window, a 3D microfluidic channel branching to three multilayered channels was successfully fabricated in a 4 mm-thick glass. In addition, appropriate design guidelines for preventing cracks in a glass and calibrating the position of the dimension of the hollow channels were studied.

본 논문에서는 심장질환 진단의 신뢰도를 높이기 위해 심전도(ECG)와 청진검사를 동시에 수행할 수 있는 휴대용 전자심전도청진기(ECGS)를 제안하였다. 심전도와 심음(HS)을 동시에 측정하기 위해 3개의 심전도 전극과 마이크 센서를 한 손에 쥘 수 있는 폭 90mm, 높이 97mm의 삼각형 모양으로 결합했다. 환자의 피부에 닿았을 때 피부 트러블을 방지하기 위해 심전도 전극으로 용량성 결합 전극을 선택했고, 마이크 센서가 있는 체스트 피스에는 실리콘 소재를 사용했다. 전극과 체스트 피스에서 측정된 신호는 각각 ECG와 HS의 주파수 대역인 0.01~100Hz와 20~250Hz의 신호만 통과시키도록 필터를 구성하였다. 필터링된 ECG 및 HS 아날로그 신호는 디지털 신호로 변환되어 무선 통신을 통해 PC로 전송되어 모니터링됩니다. HS는 이어폰을 사용하여 동시에 청진될 수 있습니다. 모니터링되는 ECG는 약 34dB의 SNR을 가지며 P-QRS-T 파형이 명확하게 보입니다. 또한 HS는 약 28dB의 SNR을 가지며 S1과 S2 모두 명확하게 보입니다. ECG 및 HS 분석에 대한 의사의 미숙함을 도울 수 있을 것으로 기대된다.

We demonstrate a two-step hybrid process for fabricating movable parts inside glass substrate using the selective laser-induced etching (SLE) process that is consisted of laser-direct writing and wet chemical etching. To obtain an influence by the optical characteristics of a glass substrate when fabricating a 3D microstructure using the SLE, we analyzed the relationship of their dimensions between the designed and the fabricated devices. Two 3D microfluidic devices are designed and fabricated on glass substrates as the demonstrations of the hybrid process: a 3D microfluidic valve device with a movable plug and a 3D microfluidic mixer with a rotatable impeller and multilayer microchannels. The valving plug and the impeller of each device are successfully moved and rotated. The smallest structure is a pillar of the impeller device, and its size is 29 μm (diameter) × 277 μm (height). We expect this study to be extended to potential applications in 3D glass microfabrication and microfluidic systems.

바이오샘플의 DNA를 대량 증폭할 수 있는 휴대형 실시간 중합효소연쇄반응(Real-time PCR) 기기에서 히터는 PCR 반응온도를 제어하기 위한 중요한 요소 중의 하나이다. 보통 빠른 히팅을 위해 소형 PCR 칩에 집적화되어 있고, 반도체 공정을이용하여 박막형태로 제작되어 PCR 칩 제작 단가가 높은 편이다. 따라서 본 연구에서는 값싸고 온도제어를 정확히 할 수 있는 히터로 칩 저항을 사용하는 것을 제안한다. 칩 저항을 사용한 히터는 구조가 단순하고 제작이 쉽다는 장점이 있다. 2.54× 2.54 cm² 크기의 실시간 PCR 칩 위에 칩 저항을 1개 또는 2개를 사용했을 때 온도분포를 시뮬레이션 하였고, 고른 온도분포를 갖는 PCR 칩을 제작했다. 또한 효율적인 PCR 칩 냉각을 위해 소형 fan이 내장된 하우징을 설계하였고, 3D 프린터로제작했다. 온도제어는 마이크로프로세서를 이용한 PID제어법(Proportional-Integral-Differential control)을 적용했다. 온도상승비와 하강비는 각각 18℃/s, 3℃/s이며, 각 PCR 반응 단계의 유지 시간을 30초로 하였을 때, 한 사이클은 약 2.66분이 걸렸고, 35 사이클은 약 93 분 내로 진행할 수 있었다.

본 연구에서는 유체를 유전체 재료로 사용하는 가변 커패시터를 시연하고, 자기 공명 현미경(MRM)의 코일에 적용 가능성을 알아보려고 하였다. 전극으로 이루어진 커패시터 구조는 마이크로 유체 채널과 통합하여 제작되었고, 커패시턴스는 채널내의 유체 충전율을 변화시키면서 측정되었다. DI water와 미네랄 오일을 사용하여 측정된 커패시턴스는 1.7 pF에서 12 pF,그리고 1.7 pF에서 2 pF으로 변하였고, 유체의 유전율에 따라 커패시턴스의 변화를 확인하였다.