• Title/Summary/Keyword: Confocal system

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Analysis of C. elegans VIG-1 Expression

  • Shin, Kyoung-Hwa;Choi, Boram;Park, Yang-Seo;Cho, Nam Jeong
    • Molecules and Cells
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    • v.26 no.6
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    • pp.554-557
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    • 2008
  • Double-stranded RNA (dsRNA) induces gene silencing in a sequence-specific manner by a process known as RNA interference (RNAi). The RNA-induced silencing complex (RISC) is a multi-subunit ribonucleoprotein complex that plays a key role in RNAi. VIG (Vasa intronic gene) has been identified as a component of Drosophila RISC; however, the role VIG plays in regulating RNAi is poorly understood. Here, we examined the spatial and temporal expression patterns of VIG-1, the C. elegans ortholog of Drosophila VIG, using a vig-1::gfp fusion construct. This construct contains the 908-bp region immediately upstream of vig-1 gene translation initiation site. Analysis by confocal microscopy demonstrated GFP-VIG-1 expression in a number of tissues including the pharynx, body wall muscle, hypodermis, intestine, reproductive system, and nervous system at the larval and adult stages. Furthermore, western blot analysis showed that VIG-1 is present in each developmental stage examined. To investigate regulatory sequences for vig-1 gene expression, we generated constructs containing deletions in the upstream region. It was determined that the GFP expression pattern of a deletion construct (${\Delta}-908$ to -597) was generally similar to that of the non-deletion construct. In contrast, removal of a larger segment (${\Delta}-908$ to -191) resulted in the loss of GFP expression in most cell types. Collectively, these results indicate that the 406-bp upstream region (-596 to -191) contains essential regulatory sequences required for VIG-1 expression.

Platelets Induce Proliferation of Human Umbilical Vein Endothelial Cells via CD154-CD40 Pathway Independently of VEGF

  • Cho, Wha-Jung;Ko, Eun-Mi;Cheon, In-Su;Jeoung, Doo-Il;Kim, Young-Myeong;Choe, Jong-Seon
    • IMMUNE NETWORK
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    • v.8 no.3
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    • pp.75-81
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    • 2008
  • Background: Platelets take part in repairing the lesions of endothelial damage. To understand the molecular mechanism of this process, we tested the hypothesis that CD154 expressed on activated platelets stimulates proliferation of human endothelial cells. Methods: The expression levels of CD154 and CD40 on platelets and endothelial cells, respectively, were measured by flow cytometry and confocal microscopy. Function-blocking monoclonal antibody against CD154 was developed after immunization with CD154-transfected L cells. Results: An anti-CD40 agonist antibody and soluble CD154 both induced significant proliferation of endothelial cells. In addition, a function-blocking anti-CD154 antibody inhibited the platelet-induced proliferation of endothelial cells, indicating that the CD154-CD40 pathway is involved in these cellular interactions. An anti-VEGF antibody failed to inhibit the proliferation. This, in addition to the fact that very small amounts of VEGF are released from platelets or endothelial cells, suggests that VEGF does not play an important role in the platelet-stimulated proliferation of endothelial cells. Conclusion: Our results indicate that platelets induce proliferation of endothelial cells by CD154-CD40 interactions independently of VEGF.

Femtosecond laser pattering of ITO film on flexible substrate (펨토초 레이저를 이용한 플렉시블 ITO 패터닝 연구)

  • Sohn, Ik-Bu;Kim, Young-Seop;Noh, Young-Chul
    • Laser Solutions
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    • v.13 no.1
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    • pp.11-15
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    • 2010
  • Indium tin oxide (ITO) provides high electrical conductivity and transparency in the visible and near IR (infrared) wavelengths. Thus, it is widely used as a transparent electrode for the fabrication of liquid crystal displays (LCDs) and organic light emitting diode displays (OLRDs), photovoltaic devices, and other optical applications. Lasers have been used for removing coating on polymer substrate for flexible display and electronic industry. In selective removal of ITO layer, laser wavelength, pulse energy, scan speed, and the repetition rate of pulses determine conditions, which are efficient for removal of ITO coating without affecting properties of the polymer substrate. ITO coating removal with a laser is more environmentally friendly than other conventional etching methods. In this paper, pattering of ITO film from polymer substrates is described. The Yb:KGW femtosecond laser processing system with a pulse duration of 250fs, a wavelength of 1030nm and a repetition rate of 100kHz was used for removing ITO coating in air. We can remove the ITO coating using a scanner system with various pulse energies and scan speeds. We observed that the amount of debris is minimal through an optical and a confocal microscope, and femtosecond laser pulses with 1030nm wavelength are effective to remove ITO coating without the polymer substrate ablation.

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Cell-Based Assay Design for High-Content Screening of Drug Candidates

  • Nierode, Gregory;Kwon, Paul S.;Dordick, Jonathan S.;Kwon, Seok-Joon
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.213-225
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    • 2016
  • To reduce attrition in drug development, it is crucial to consider the development and implementation of translational phenotypic assays as well as decipher diverse molecular mechanisms of action for new molecular entities. High-throughput fluorescence and confocal microscopes with advanced analysis software have simplified the simultaneous identification and quantification of various cellular processes through what is now referred to as high-content screening (HCS). HCS permits automated identification of modifiers of accessible and biologically relevant targets and can thus be used to detect gene interactions or identify toxic pathways of drug candidates to improve drug discovery and development processes. In this review, we summarize several HCS-compatible, biochemical, and molecular biology-driven assays, including immunohistochemistry, RNAi, reporter gene assay, CRISPR-Cas9 system, and protein-protein interactions to assess a variety of cellular processes, including proliferation, morphological changes, protein expression, localization, post-translational modifications, and protein-protein interactions. These cell-based assay methods can be applied to not only 2D cell culture but also 3D cell culture systems in a high-throughput manner.

Development of Elimination Method of Measurement noise to Improve accuracy for White Light Interferometry (백색광 간섭계의 정밀도 향상을 위한 노이즈 제거 방법)

  • Ko, Kuk-Won;Cho, Soo-Yong;Kim, Min-Young
    • Journal of Institute of Control, Robotics and Systems
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    • v.14 no.6
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    • pp.519-522
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    • 2008
  • As industry of a semiconductor and LCD industry have been rapidly growing, precision technologies of machining such as etching and 3D measurement are required. Stylus has been important measuring method in traditional manufacturing process. However, its disadvantages are low measuring speed and damage possibility at contacting point. To overcome mentioned disadvantage, non-contacting measurement method is needed such as PMP(Phase Measuring Profilometry), WSI(white scanning interferometer) and Confocal Profilometry. Among above 3 well-known methods, WSI started to be applied to FPD(flat panel display) manufacturing process. Even though it overcomes 21t ambiguity of PMP method and can measure objects which has specular surface, the measuring speed and vibration coming from manufacturing machine are one of main issue to apply full automatic total inspection. In this study, We develop high speed WSI system and algorithm to reduce unknown noise. The developing WSI and algorithm are implemented to measure 3D surface of wafer. Experimental results revealed that the proposed system and algorithm are able to measure 3D surface profile of wafer with a good precision and high speed.

Expression of CD320 in human B cells in addition to follicular dendritic cells

  • Cho, Wha-Jung;Choi, Jin-Suk;Park, Chan-Hum;Yoon, Sun-Ok;Jeoung, Doo-Il;Kim, Young-Myeong;Choe, Jong-Seon
    • BMB Reports
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    • v.41 no.12
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    • pp.863-867
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    • 2008
  • CD320 has been recently discovered and reported as a follicular dendritic cell (FDC) protein. Although CD320 is known to enhance proliferation of germinal center (GC) B cells, little other information is available. In this study, we investigated its cellular distribution in the GC. Confocal microscopy of human tonsil sections revealed co-localization of CD320 with CD19 and CD38 but not with CD3 indicating that GC B cells expressed CD320 in addition to FDC. In purified GC B cells, CD320 expression was inhibited in the nucleus, membrane and cytoplasm. Reverse transcriptase-polymerase chain reaction confirmed CD320 mRNA expression in B cells. These finding indicate that CD320 is expressed in B cells in addition to FDC, and that its GC activity may be more complicated than previously thought.

Apoptin gene delivery by a PAMAM dendrimer modified with a nuclear localization signal peptide as a gene carrier for brain cancer therapy

  • Bae, Yoonhee;Lee, Jell;Kho, Changwon;Choi, Joon Sig;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.5
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    • pp.467-478
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    • 2021
  • In this study, we aimed to synthesize PAMAMG3 derivatives (PAMAMG3-KRRR and PAMAMG3-HKRRR), using KRRR peptides as a nuclear localization signal and introduced histidine residues into the KRRR-grafted PAMAMG3 for delivering a therapeutic, carcinoma cell-selective apoptosis gene, apoptin into human primary glioma (GBL-14) cells and human dermal fibroblasts. We examined their cytotoxicity and gene expression using luciferase activity and enhanced green fluorescent protein PAMAMG3 derivatives in both cell lines. We treated cells with PAMAMG3 derivative/apoptin complexes and investigated their intracellular distribution using confocal microscopy. The PAMAMG3-KRRR and PAMAMG3-HKRRR dendrimers were found to escape from endolysosomes into the cytosol. The JC-1 assay, glutathione levels, and Annexin V staining results showed that apoptin triggered cell death in GBL-14 cells. Overall, these findings indicated that the PAMAMG3-HKRRR/apoptin complex is a potential candidate for an effective nonviral gene delivery system for brain tumor therapy in vitro.

Optomechanical Design and Structural Analysis of Linear Astigmatism Free - Three Mirror System Telescope for CubeSat and Unmanned Aerial Vehicle

  • Han, Jimin;Lee, Sunwoo;Park, Woojin;Moon, Bongkon;Kim, Geon Hee;Lee, Dae-Hee;Kim, Dae Wook;Pak, Soojong
    • The Bulletin of The Korean Astronomical Society
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    • v.46 no.1
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    • pp.38.3-38.3
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    • 2021
  • We are developing an optomechanical design of infrared telescope for the CubeSat and Unmanned Aerial Vehicle (UAV) which adapts the Linear Astigmatism Free- Three Mirror System in the confocal off-axis condition. The small entrance pupil (diameter of 40 mm) and the fast telescope (f-number of 1.9) can survey large areas. The telescope structure consists of three mirror modules and a sensor module, which are assembled on the base frame. The mirror structure has duplex layers to minimize a surface deformation and physical size of a mirror mount. All the optomechanical parts and three freeform mirrors are made from the same material, i.e., aluminum 6061-T6. The Coefficient of Thermal Expansion matching single material structure makes the imaging performance to be independent of the thermal expansion. We investigated structural characteristics against external loads through Finite Element Analysis. We confirmed the mirror surface distortion by the gravity and screw tightening, and the overall contraction/expansion following the external temperature environment change (from -30℃ to +30℃).

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Feasibility Study for the Development of a Device for Pathological Tissue (병리학적 조직 진단장치 개발에 대한 타당성 분석 연구)

  • Ko Chea-Ok;Park Min-Young;Kim Jeong-Lan;Lee Ae-Kyoung;Choi Hyung-Do;Choi Jae-Ic;Pack Jeong-Ki
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.17 no.4 s.107
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    • pp.341-350
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    • 2006
  • In this paper, a new method for detecting breast cancer is proposed, which utilizes dielectric characteristics of pathological tissues and time delay of back scattered response, and its feasibility was investigated. We have developed a detection algorithm and verified it by numerical simulation and measurement for a prototype system. For a prototype system, we have fabricated experimental model(artificial breast with a cancer) and UWB(ultra-wideband) antenna. The results of the measurement simulation show an excellent detection capability of a cancer tissue. It is found that a good UWB antenna and a good calibration signal are key elements of such detection system. Further study is ongoing to develop a commercial system.

Development and evaluation of probiotic delivery systems using the rennet-induced gelation of milk proteins

  • Ha, Ho-Kyung;Hong, Ji-Young;Ayu, Istifiani Lola;Lee, Mee-Ryung;Lee, Won-Jae
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.1182-1193
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    • 2021
  • The aims of this study were to develop a milk protein-based probiotic delivery system using a modified rennet-induced gelation method and to determine how the skim milk powder concentration level and pH, which can affect the rennet-induced intra- and inter-molecular association of milk proteins, affect the physicochemical properties of the probiotic delivery systems, such as the particle size, size distribution, encapsulation efficiency, and viability of probiotics in simulated gastrointestinal tract. To prepare a milk protein-based delivery system, skim milk powder was used as a source of milk proteins with various concentration levels from 3 to 10% (w/w) and rennet was added to skim milk solutions followed by adjustment of pH from 5.4 or 6.2. Lactobacillus rhamnosus GG was used as a probiotic culture. In confocal laser scanning microscopic images, globular particles with a size ranging from 10 ㎛ to 20 ㎛ were observed, indicating that milk protein-based probiotic delivery systems were successfully created. When the skim milk powder concentration was increased from 3 to 10% (w/w), the size of the delivery system was significantly (p < 0.05) increased from 27.5 to 44.4 ㎛, while a significant (p < 0.05) increase in size from 26.3 to 34.5 ㎛ was observed as the pH was increased from 5.4 to 6.4. An increase in skim milk powder concentration level and a decrease in pH led to a significant (p < 0.05) increase in the encapsulation efficiency of probiotics. The viability of probiotics in a simulated stomach condition was increased when probiotics were encapsulated in milk protein-based delivery systems. An increase in the skim milk powder concentration and a decrease in pH resulted in an increase in the viability of probiotics in simulated stomach conditions. It was concluded that the protein content by modulating skim milk powder concentration level and pH were the key manufacturing variables affecting the physicochemical properties of milk protein-based probiotic delivery systems.