• Title/Summary/Keyword: Fluorescence sensing

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Intracellular Electrical Stimulation on PC-12 Cells through Vertical Nanowire Electrode

  • Kim, Hyungsuk;Kim, Ilsoo;Lee, Jaehyung;Lee, Hye-young;Lee, Eungjang;Jeong, Du-Won;Kim, Ju-Jin;Choi, Heon-Jin
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.407-407
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    • 2014
  • Nanotechnology, especially vertically grown silicon nanowires, has gotten great attentions in biology due to characteristics of one dimensional nanostructure; controllable synthetic structure such as lengths, diameters, densities. Silicon nanowires are promising materials as nanoelectrodes due to their highly complementary metal-oxide-semiconductor (CMOS) - and bio-compatibility. Silicon nanowires are so intoxicated that are effective for bio molecular delivery and electrical stimulation. Vertical nanowires with integrated Au tips were fabricated for electrical intracellular interfacing with PC-12 cells. We have made synthesized two types of nanowire devices; one is multi-nanowires electrode for bio molecular sensing and electrical stimulation, and the other is single-nanowires electrode respectively. Here, we demonstrate that differentiation of Nerve Growth Factor (NGF) treated PC-12 cells can be promoted depending on different magnitudes of electrical stimulation and density of Si NWs. It was fabricated by both bottom-up and top-down approaches using low pressure chemical vapor deposition (LPCVD) with high vacuuming environment to electrically stimulate PC-12 cells. The effects of electrical stimulation with NGF on the morphological differentiation are observed by Scanning Electron Microscopy (SEM), and it induces neural outgrowth. Moreover, the cell cytosol can be dyed selectively depending on the degree of differentiation along with fluorescence microscopy measurement. Vertically grown silicon nanowires have further expected advantages in case of single nanowire fabrication, and will be able to expand its characteristics to diverse applications.

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Evaluation of Photochemical Reflectance Index (PRI) Response to Soybean Drought stress under Climate Change Conditions (기후변화 조건에서 콩 한발스트레스에 대한 광화학 반사 지수 반응 평가)

  • Sang, Wan-Gyu;Kim, Jun-Hwan;Shin, Pyeong;Baek, Jae-Kyeong;Lee, Yun-Ho;Cho, Jung-Il;Seo, Myung-Chul
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.21 no.4
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    • pp.261-268
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    • 2019
  • Climate change and drought stress are having profound impacts on crop growth and development by altering crop physiological processes including photosynthetic activity. But finding a rapid, efficient, and non-destructive method for estimating environmental stress responses in the leaf and canopy is still a difficult issue for remote sensing research. We compared the relationships between photochemical reflectance index(PRI) and various optical and experimental indices on soybean drought stress under climate change conditions. Canopy photosynthesis trait, biomass change, chlorophyll fluorescence(Fv/Fm), stomatal conductance showed significant correlations with midday PRI value across the drought stress period under various climate conditions. In high temperature treatment, PRI were more sensitive to enhanced drought stress, demonstrating the negative effect of the high temperature on the drought stress. But high CO2 concentration alleviated the midday depression of both photosynthesis and PRI. Although air temperature and CO2 concentration could affect PRI interpretation and assessment of canopy radiation use efficiency(RUE), PRI was significantly correlated with canopy RUE both under climate change and drought stress conditions, indicating the applicability of PRI for tracking the drought stress responses in soybean. However, it is necessary to develop an integrated model for stress diagnosis using PRI at canopy level by minimizing the influence of physical and physiological factors on PRI and incorporating the effects of other vegetation indices.

Raloxifene Induces Autophagy-Dependent Cell Death in Breast Cancer Cells via the Activation of AMP-Activated Protein Kinase

  • Kim, Dong Eun;Kim, Yunha;Cho, Dong-Hyung;Jeong, Seong-Yun;Kim, Sung-Bae;Suh, Nayoung;Lee, Jung Shin;Choi, Eun Kyung;Koh, Jae-Young;Hwang, Jung Jin;Kim, Choung-Soo
    • Molecules and Cells
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    • v.38 no.2
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    • pp.138-144
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    • 2015
  • Raloxifene is a selective estrogen receptor modulator (SERM) that binds to the estrogen receptor (ER), and exhibits potent anti-tumor and autophagy-inducing effects in breast cancer cells. However, the mechanism of raloxifene-induced cell death and autophagy is not well-established. So, we analyzed mechanism underlying death and autophagy induced by raloxifene in MCF-7 breast cancer cells. Treatment with raloxifene significantly induced death in MCF-7 cells. Raloxifene accumulated GFP-LC3 puncta and increased the level of autophagic marker proteins, such as LC3-II, BECN1, and ATG12-ATG5 conjugates, indicating activated autophagy. Raloxifene also increased autophagic flux indicators, the cleavage of GFP from GFP-LC3 and only red fluorescence-positive puncta in mRFP-GFP-LC3-expressing cells. An autophagy inhibitor, 3-methyladenine (3-MA), suppressed the level of LC3-II and blocked the formation of GFP-LC3 puncta. Moreover, siRNA targeting BECN1 markedly reversed cell death and the level of LC3-II increased by raloxifene. Besides, raloxifene-induced cell death was not related to cleavage of caspases-7, -9, and PARP. These results indicate that raloxifene activates autophagy-dependent cell death but not apoptosis. Interestingly, raloxifene decreased the level of intracellular adenosine triphosphate (ATP) and activated the AMPK/ULK1 pathway. However it was not suppressed the AKT/mTOR pathway. Addition of ATP decreased the phosphorylation of AMPK as well as the accumulation of LC3-II, finally attenuating raloxifene-induced cell death. Our current study demonstrates that raloxifene induces autophagy via the activation of AMPK by sensing decreases in ATP, and that the overactivation of autophagy promotes cell death and thereby mediates the anti-cancer effects of raloxifene in breast cancer cells.

Analysis of Human Serum Amyloid A-1 Concentrations Using a Lateral Flow Immunoassay with CdSe/ZnS Quantum Dots (Human Serum Amyloid A-1 단백질 농도 분석을 위한 CdSe/ZnS 양자점 기반의 Lateral Flow Immunoassay 방법 개발)

  • Fajri, Aidil;Goh, Eunseo;Lee, Sanghyuk;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.30 no.4
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    • pp.429-434
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    • 2019
  • A lateral flow immunoassay platform utilizing antibody functionalized water soluble CdSe/ZnS semiconductor quantum dots (QDs) was developed for the analysis of human serum amyloid A-1 (hSAA1) in a buffer solution. hSAA1 was chosen as a target protein because it is regarded as a potential biomarker associated with early diagnosis and prognosis in patients of lung cancer. The immunoassay strip on a nitrocellulose membrane was fabricated by spraying two lines composed of a test line with a monoclonal antibody against hSAA1 (10G1) (anti hSAA1) and a control line of anti-chicken IgY. While the CdSe/ZnS QDs synthesized in an organic phase were transferred to a water phase by ligand exchange using carboxylic acid modified alkane thiol. The QDs was then conjugated to monoclonal antibody against hSAA1 (14F8) [anti hSAA1 (14F8)] and used as a fluorescent detection probe. The sequential lateral flow of hSAA1 in different concentration and QDs-anti hSAA1 (14F8) complex allowed to form the surface sandwich complex of anti hSAA1 (10G1)/hSAA1/QD-anti hSAA1 (14F8), which was then analyzed using fluorescence microscope. A 100 nM concentration of hSAA1 protein can be detected by naked eyes under an optimized lateral flow buffer condition with a sensing time of 5 mins.

The spy-gfp Operon Fusion in Salmonella Enteritidis and Salmonella Gallinarum Senses the Envelope Stress (Salmonella Enteritidis와 Salmonella Gallinarum의 세균막 스트레스를 인식하는 spy-gfp 오페론 융합)

  • Kang, Bo Gyeong;Bang, Iel Soo
    • Journal of Dairy Science and Biotechnology
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    • v.36 no.4
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    • pp.208-219
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    • 2018
  • Emergence of drug resistant strains of Salmonella enterica threatens milk processing and related dairy industries, thereby increasing the need for development of new anti-bacterials. Developments of antibacterial drugs are largely aimed to target the bacterial envelope, but screening their efficacy on bacterial envelope is laborious. This study presents a potential biosensor for envelope-specific stress in which a gfp reporter gene fused to spy gene encoding a periplasmic chaperone protein Spy (spheroplast protein y) that can sense envelope stress signals transduced by two major two-component signal transduction systems BaeSR and CpxAR in Salmonella enterica serovars Enteritidis and S. Gallinarum. Using spy-gfp operon fusions in S. Enterititis and S. Gallinarum, we found that spy transcription in both serovars was greatly induced when Salmonella cells were forming the spheroplast and were treated with ethanol or a membrane-disrupting antibiotic polymyxin B. These envelope stress-specific inductions of spy transcription were abrogated in mutant Salmonella lacking either BaeR or CpxR. Results illustrate that induction of Spy expression can be efficiently triggered by two-component signal transduction systems sensing envelope stress conditions, and thereby suggest that monitoring the spy transcription by spy-gfp operon fusions would be helpful to determine if developing antimicrobials can damage envelopes of S. Enteritidis and S. Gallinarum.

Non-contact Stress Measurement in Steel Member of PSC Box Bridge Using Raman Spectroscopy (라만 형광 분광법을 이용한 PSC 박스교 인장케이블 응력측정방법 연구)

  • Kim, Jongwoo;Kim, Namgyu
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.23 no.2
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    • pp.130-134
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    • 2019
  • In this paper, a laser-based non-contact load cell is newly developed for measuring forces in prestressed concrete tendons. First, alumina particles have been sprayed onto an empty load cell which has no strain gauges on it, and the layer has been used as a passive stress sensor. Then, the spectral shifts in fluorescence spectroscopy have been measured using a laser-based spectroscopic system under various force levels, and it has been found that the relation of applied force and spectral shift is linear in a lab-scale test. To validate the field applicability of the customized load cell, a full-scale prestressed concrete specimen has been constructed in a yard. During the field test, it was, however, found that the coating surface has irregular stress distribution. Therefore, the location of a probe has to be fixed onto the customized load cell for using the coating layer as a passive stress sensor. So, a prototype customized load cell has been manufactured, which consists of a probe mount on its casing. Then, by performing lab-scale uniaxial compression tests with the prototype load cell, a linear relation between compression stress and spectrum shift at a specific point where laser light had been illuminated has been detected. Thus, it has a high possibility to use the prototype load cell as a force sensor of prestressed concrete tendons.

Prelaunch Study of Validation for the Geostationary Ocean Color Imager (GOCI) (정지궤도 해색탑재체(GOCI) 자료 검정을 위한 사전연구)

  • Ryu, Joo-Hyung;Moon, Jeong-Eon;Son, Young-Baek;Cho, Seong-Ick;Min, Jee-Eun;Yang, Chan-Su;Ahn, Yu-Hwan;Shim, Jae-Seol
    • Korean Journal of Remote Sensing
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    • v.26 no.2
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    • pp.251-262
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    • 2010
  • In order to provide quantitative control of the standard products of Geostationary Ocean Color Imager (GOCI), on-board radiometric correction, atmospheric correction, and bio-optical algorithm are obtained continuously by comprehensive and consistent calibration and validation procedures. The calibration/validation for radiometric, atmospheric, and bio-optical data of GOCI uses temperature, salinity, ocean optics, fluorescence, and turbidity data sets from buoy and platform systems, and periodic oceanic environmental data. For calibration and validation of GOCI, we compared radiometric data between in-situ measurement and HyperSAS data installed in the Ieodo ocean research station, and between HyperSAS and SeaWiFS radiance. HyperSAS data were slightly different in in-situ radiance and irradiance, but they did not have spectral shift in absorption bands. Although all radiance bands measured between HyperSAS and SeaWiFS had an average 25% error, the 11% absolute error was relatively lower when atmospheric correction bands were omitted. This error is related to the SeaWiFS standard atmospheric correction process. We have to consider and improve this error rate for calibration and validation of GOCI. A reference target site around Dokdo Island was used for studying calibration and validation of GOCI. In-situ ocean- and bio-optical data were collected during August and October, 2009. Reflectance spectra around Dokdo Island showed optical characteristic of Case-1 Water. Absorption spectra of chlorophyll, suspended matter, and dissolved organic matter also showed their spectral characteristics. MODIS Aqua-derived chlorophyll-a concentration was well correlated with in-situ fluorometer value, which installed in Dokdo buoy. As we strive to solv the problems of radiometric, atmospheric, and bio-optical correction, it is important to be able to progress and improve the future quality of calibration and validation of GOCI.