• 대한기계학회논문집B
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• 제30권2호
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• pp.153-160
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• 2006

A technique for measuring surface temperature field in micro scale is newly proposed, which uses temperature-sensitive fluorescent (TSF) dye coated on the surface and is easily implemented with a fluorescence microscope and a CCD camera. The TSF dye is chosen among mixtures of various chemical compositions including rhodamine B as the fluorescent dye to be most sensitive to temperature change. In order to examine the effectiveness of this temperature measurement technique, numerical analysis and experiment on transient conduction heat transfer for two different substrate materials, i. e., silicon and glass, are performed. In the experiment, to accurately measure the temperature with high resolution temperature calibration curves were obtained with very fine spatial units. The experimental results agree qualitatively well with the numerical data in the silicon and glass substrate cases so that the present temperature measurement method proves to be quite reliable. In addition, it is noteworthy that the glass substrate is more appropriate to be used as thermally-insulating locally-heating heater in micro thermal devices. This fact is identified in the temperature measuring experiment on the locally-heating heaters made on the wafer of silicon and glass substrates. Accordingly, this technique is capable of accurate and non-intrusive high-resolution measurement of temperature field in microscale.

• BMB Reports
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• 제50권2호
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• pp.55-57
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• 2017

Toll-like receptor 4 (TLR4) together with MD2, one of the key pattern recognition receptors for a pathogen-associated molecular pattern, activates innate immunity by recognizing lipopolysaccharide (LPS) of Gram-negative bacteria. Although LBP and CD14 catalyze LPS transfer to the TLR4/MD2 complex, the detail mechanisms underlying this dynamic LPS transfer remain elusive. Using negative-stain electron microscopy, we visualized the dynamic intermediate complexes during LPS transfer-LBP/LPS micelles and ternary CD14/LBP/LPS micelle complexes. We also reconstituted the entire cascade of LPS transfer to TLR4/MD2 in a total internal reflection fluorescence (TIRF) microscope for a single molecule fluorescence analysis. These analyses reveal longitudinal LBP binding to the surface of LPS micelles and multi-round binding/unbinding of CD14 to single LBP/LPS micelles via key charged residues on LBP and CD14. Finally, we reveal that a single LPS molecule bound to CD14 is transferred to TLR4/MD2 in a TLR4-dependent manner. These discoveries, which clarify the molecular mechanism of dynamic LPS transfer to TLR4/MD2 via LBP and CD14, provide novel insights into the initiation of innate immune responses.

• 한국가시화정보학회지
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• 제13권1호
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• pp.35-42
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• 2015

CD4+ T-cell count determines the effectiveness for antiretroviral therapy (ART) in patients with human immunodeficiency virus (HIV). Although ART slows the progression of HIV to AIDS, rapid counting of CD4+ T lymphocytes with a drop of patient's blood sample is urgently needed to ensure timely ART treatment in rural areas. Recently point-of-care CD4 testing devices have been developed by using non-flow based imaging cytometer incorporated with a sample cartridge where CD4+ T cells are reacted with fluorescently tagged specific antibodies. Here we conducted an experimental study using a conventional fluorescence microscope-based imaging system to quantitate the interaction of CD4 antibodies with CD4+ T cells at different reaction conditions. We demonstrated that a fast and affordable point-of-care CD4 test is feasible with a far less amount of antibodies and a shorter incubation time compared with a conventional sample preparation protocol for flow cytometry. We also proposed a general method to evaluate and compare the detection limit across different CD4 counting platforms by using fluorescently labelled microbeads for intensity calibration.

• 한국정밀공학회지
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• 제25권5호
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• pp.140-147
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• 2008

In this study, a carbon nanotube probe (CNT probe) is proposed as a mechanical force transducer for the measurement of pico-Newton (pN) order force in biological applications. In order to measure nantube's displacement in the air or liquid environment, the fabrication of a CNT probe with tip-specific loading of fluorescent dyes is performed using tip- specific functionalization of the nanotube and chemical bonding between dyes and nanotube. Also, we experimentally investigated the mechanical properties of the CNT probe using electrostatic actuation and fluorescence microscope measurement. Using fluorescence measurement of the tip deflection according to the applied voltage, we optimized the bending stiffness of the CNT probe, therefore determined the spring constant of the CNT probe. The results show that the spring constant of CNT probes is as small as 1 pN/nm and CNT probes can be used to measure pN order force.

• International Journal of Industrial Entomology and Biomaterials
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• 제15권1호
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• pp.35-38
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• 2007

In this study, ORF 46 of Bombyx mod nucleopolyhedrovirus(Bm46) fused with EGFP was expressed in Bombyx mod cell lines, larvae and pupae by BmNPV Bacmid system. Bm46 and EGFP were cloned into donor plasmid pFastBacHTb, which was transformed to competent DH10B cells containing helper and BmNPV bacmid by site-specific transposition. Recombinant bacmid was used to transfected BmN-4 cells to produce the recombinant baculovirus vBm-Bm46-EGFP. Recombination virus was injected into silkworm larvae and pupae. The expression of the fusion protein was monitored by examining green fluorescence using a fluorescent microscope. Intense fluorescence in cells and silkworm was observed at 4 days post-infection, indicating the Bm46-EGFP fusion gene was expressed successfully.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.859-862
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• 2001

For the development of preferable immunosensor and protein chip, the viologen Langmuir-Blodgett (LB) multilayer was fabricated on the surface, and then protein A was adsorbed on the proposed viologen LB film by electrostatic attractive force. The Immunoglobulin G (IgG) labeled with fluorescence marker was self-assembled on the fabricated protein A film. The topographies of the deposited films were investigated by using atomic force microscope (AFM). The immobilization of IgG was verified by fluorescence spectrum. Such structures can be used as sublayers for various kinds of IgG immobilization toward immunosensors and protein chip.

• 한국가시화정보학회지
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• 제10권2호
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• pp.25-31
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• 2012

High-performance smartphones, equipped with a digital camera and an application software, can render conventional bench-top laboratory instruments mobile at affordable costs. As the smartphone-based devices are portable and wireless, they have wide applications especially in providing point-of-care (POC) tests in resource-constrained areas. We developed a hand-held diagnostic system, Mobile SmartScope, which consists of a small optical unit integrated with a smartphone. The performance of the SmartScope was favorably compared with that of conventional light microscopy in detecting and quantifying red blood cells. We also evaluated the fluorescence detection limit of the SmartScope incorporated with a blue light-emitting diode and appropriate optical filters by using fluorescently labeled microbeads for intensity calibration.

• Journal of Photoscience
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• 제11권2호
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• pp.47-53
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• 2004

A laser scanning fluorescence microscope system has been fabricated for single molecule detection (SMD). Problems associated with the system set-up have been discussed along with proper suggestions. Based on the SMD results obtained by using the apparatus, a statistical method has been suggested to determine the minimum number of required molecules to form a group of uniform average in a selected error range.

• 한국가시화정보학회:학술대회논문집
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• 한국가시화정보학회 2004년도 Proceedings of 2004 Korea-Japan Joint Seminar on Particle Image Velocimetry
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• pp.91-110
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• 2004

A comprehensive three-dimensional nano-particle tracking technique in micro- and nano-scale spatial resolution using the Total Internal Reflection Fluorescence Microscope (TIRFM) is discussed. Evanescent waves from the total internal reflection of a 488nm argon-ion laser are used to measure the hindered Brownian diffusion within few hundred nanometers of a glass-water interface. 200-nm fluorescence-coated polystyrene spheres are used as tracers to achieve three-dimensional tracking within the near-wall penetration depth. A novel ratiometric imaging technique coupled with a neural network model is used to tag and track the tracer particles. This technique allows for the determination of the relative depth wise locations of the particles. This analysis, to our knowledge is the first such three-dimensional ratiometric nano-particle tracking velocimetry technique to be applied for measuring Brownian diffusion close to the wall.

• 한국광학회지
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• 제8권1호
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• pp.42-46
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• 1997

단일 색소 분자가 공초점 현미경의 검출 영역을 통과하면서 방출하는 형광폭발신호를 수집하기 위해서 컴퓨터 인터페이싱 광자계수기를 제작하였다. 사용한 계수기 소자들에 의해서 결정되는 최대 계수 속도는 약 80 MHz이며, 486급 PC 컴퓨터와 BASIC으로 작성한 프로그램을 사용하였을 때 최소 bin-width는 약 25.mu.s이었다. 이 계수기를 사용하여 에틸렌글리콜 용액내에 있는 JA22 단일 분자의 형광폭발신호를 수집하였으며 그 특성을 간략하게 기술하였다. 이때 색소 분자의 농도는 약 1 * $10^{-11}$ mol/L이었다.