• Title/Summary/Keyword: Fluorescent properties

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Intergeneric Protoplast Fusion between Rhizopus oryzae and Aspergillus oryzae (Rhizopus oryzae와 Aspergillus oryzae의 속간 원형질체융합)

  • Lee, Soo-Youn;Jung, Sung-Won;Kim, Seong-Han;Lee, Yung-Nok
    • Korean Journal of Microbiology
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    • v.31 no.3
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    • pp.218-223
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    • 1993
  • Conditions for the release and regeneration of protoplasts form Rhizopus oryzae and intergeneric protoplast fusion between Rhizopus oryzae and Aspergillus oryzae were studied. High yields of protoplast fusion between Rhizopus oryzae and Aspergillus oxyzae were studied. High yield of protoplasts from young germilings of R. oryzae were obtained by using lytic enzymes containing chitosanase (3 mg/ml), chitinase (3 mg/ml) and Novozym 234 (5 mg/ml). 0.5M glucose was used as the osmotic stabilizer and optimum pH of buffer was determined to be pH 7.5-8.0. Under these conditions, protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts regenerated on solid medium with a soft agar overlay. We have also carried out protoplasts fusion between R. oryzae and A. oryzae and have succeeded in obtaining three types of intergeneric fusants. In these experiments, 35% PEG-4000 and 10 mM CaCl$_{2}$ were used as fsogenic agents, and auxotrophic properties were used as a genetic marker to select fusants. Complementation frequency be protoplasts fusion of A. oxyzae and R. oryzae was 4.4% * 10$^{-5}$ . The fusant strains of the first type were prototrophs showing an Aspergillus type morphology with dark-yellow sporulation, those of the second type were also Apergillus type morphology but showed no sporulation. And the strains of the third type stopped growing when fusion products grown on regeneration minimal medium were transferred to fresh minimal medium. The formation of fusion products was observed by fluorescent vital stains for complementary labelling of protoplats from R. oryzae and A. oryzae. Rhodamine 6G and fluorescein diacetate wer useful complementary vital stains of Rhizopus and Aspergillus protoplasts for visualization of requency and type (dicell, multicell) of fusion.

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Techniques for Evaluation of LAMP Amplicons and their Applications in Molecular Biology

  • Esmatabadi, Mohammad javad Dehghan;Bozorgmehr, Ali;zadeh, Hesam Motaleb;Bodaghabadi, Narges;Farhangi, Baharak;Babashah, Sadegh;Sadeghizadeh, Majid
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7409-7414
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    • 2015
  • Loop-mediated isothermal amplification (LAMP) developed by Notomi et al. (2000) has made it possible to amplify DNA with high specificity, efficiency and rapidity under isothermal conditions. The ultimate products of LAMP are stem-loop structures with several inverted repeats of the target sequence and cauliflower-like patterns with multiple loops shaped by annealing between every other inverted repeats of the amplified target in the similar strand. Because the amplification process in LAMP is achieved by using four to six distinct primers, it is expected to amplify the target region with high selectivity. However, evaluation of reaction accuracy or quantitative inspection make it necessary to append other procedures to scrutinize the amplified products. Hitherto, various techniques such as turbidity assessment in the reaction vessel, post-reaction agarose gel electrophoresis, use of intercalating fluorescent dyes, real-time turbidimetry, addition of cationic polymers to the reaction mixture, polyacrylamide gel-based microchambers, lateral flow dipsticks, fluorescence resonance energy transfer (FRET), enzyme-linked immunosorbent assays and nanoparticle-based colorimetric tests have been utilized for this purpose. In this paper, we reviewed the best-known techniques for evaluation of LAMP amplicons and their applications in molecular biology beside their advantages and deficiencies. Regarding the properties of each technique, the development of innovative prompt, cost-effective and precise molecular detection methods for application in the broad field of cancer research may be feasible.

Red Emission Properties of Organic EL Having Hole Blocking Layer (정공블록킹층을 설치한 유기 EL의 적색발광특성)

  • Kim, Hyeong-Gweon;Lee, Eun-Hak
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.37 no.6
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    • pp.17-23
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    • 2000
  • In this study, we prepared red organic light-emitting-diode(OLED) with a fluorescent dye(Sq)-doped and inserted between emission and cathode layer 1,3-bis(5-p-t-butylphenyl)-1,3,4-oxadiazol-2-yl)benzene (OXD7) or/and tris(8-hydroxyquinoline) aluminum ($Alq_3$) layers for increasing electroluminescent(EL) efficiency. This inserting effect has been observed and EL mechanism characteristics have been examined. The hole transfer layer is a N,N'-diphenyl-N,N'-bis-(3-methyl phenyl)-1,1'-diphenyl-4,4'-diamine (TPD), and the host and guest materials of emission layer is $Alq_3$ and bis[1-methyl-3,3'-dimethyl-2-indorindiylmethyl] squaraine (Sq), respectively. For the inserting of $Alq_3$, emission efficiency increased. But we can not obtained highly pure red emission owing to the emission of inserting $Alq_3$ layer. The inserting of OXD7 makes hole block and accumulate. Because of increasing recombination probability of electron and hole, highly pure red color can be held. Simultaneously brightness characteristics and emission efficiency could improve.

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Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

  • Chowdhury, Salina A.;Lim, Man-Ho
    • Bulletin of the Korean Chemical Society
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    • v.32 no.2
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    • pp.583-589
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    • 2011
  • Fluorescence correlation spectroscopy (FCS) is a sophisticated and an accurate analytical technique used to study the diffusion of molecules in a solution at the single-molecule level. FCS is strongly affected by many factors such as the stability of the excitation power, photochemical processes, mismatch between the refractive indices, and variations in the cover glass thickness. We have studied FCS near the surface of a cover glass by using rhodamine 123 as a fluorescent probe and have observed that the surface has a strong influence on the measurements. The temporal autocorrelation of FCS decays with two characteristic times when the confocal detection volume is positioned near the surface of the cover glass. As the position of the detection volume is moved away from the surface, the FCS autocorrelation becomes one-component decaying; the characteristic time of the decay is the same as the faster-decaying component in the FCS autocorrelation near the surface. This observation suggests that the faster component can be attributed to the free diffusion of the probe molecules in the solution, while the slow component has its origin from the interaction between the probe molecules and the surface. We have characterized the surface contribution to the FCS measurements near the surface by changing the position of the detection volume relative to the surface. The influence of the surface on the diffusion of the probe molecules was monitored by changing the chemical properties of the surface. The surface contribution to the temporal autocorrelation of the FCS strongly depends on the chemical nature of the surface. The hydrophobicity of the surface is a major factor determining the surface influence on the free diffusion of the probe molecules near the surface.

The Effect of Various Process Conditions on the Physical Properties of Dense Silver Films, Prepared by Using Sputter Deposition on Polyester Substrate (Polyester 상에서 Sputter 증착되는 고 밀도 은경 박막의 물리적 특성에 미치는 공정조건 변화의 효과)

  • Ri, Ui-Jae;Hwang, Tae-Su
    • Korean Journal of Materials Research
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    • v.9 no.7
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    • pp.707-714
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    • 1999
  • To save electrical energy as much as 40 % for fluorescent lighting, the reflectors coated with silver reflective thin films recently became popular with higher reflectivities and long life. The thin films fabricated by using sputtering techniques are produced mainly in U.S.A. On the other hand, some silver films deposited by using evaporation methods show low adhesion in general, although the reflectivity is no problem. We have studied various PVD methods to obtain thin films with high reflectivity and adhesion on a substrate of polyester, for a couple of years. Silver films manufactured byusing evaporation showed the reflectivity of 96.4 % and the adhesion of $12 kg/\textrm{cm}^2$. while samples manufactured by using sputtering depicted the adhesion as much as $20 Kg/\textrm{cm}^2$ that is almost double, although their reflectivity was not much different. X-ray diffraction spectra for the sputtered films demonstrated a preferential growth on (111) plane and the cross-sections of the specimens revealed a dense columnar structure to result in the enhanced adhesion.

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Two-Cell Spheroid Angiogenesis Assay System Using Both Endothelial Colony Forming Cells and Mesenchymal Stem Cells

  • Shah, Sajita;Kang, Kyu-Tae
    • Biomolecules & Therapeutics
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    • v.26 no.5
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    • pp.474-480
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    • 2018
  • Most angiogenesis assays are performed using endothelial cells. However, blood vessels are composed of two cell types: endothelial cells and pericytes. Thus, co-culture of two vascular cells should be employed to evaluate angiogenic properties. Here, we developed an in vitro 3-dimensional angiogenesis assay system using spheroids formed by two human vascular precursors: endothelial colony forming cells (ECFCs) and mesenchymal stem cells (MSCs). ECFCs, MSCs, or ECFCs+MSCs were cultured to form spheroids. Sprout formation from each spheroid was observed for 24 h by real-time cell recorder. Sprout number and length were higher in ECFC+MSC spheroids than ECFC-only spheroids. No sprouts were observed in MSC-only spheroids. Sprout formation by ECFC spheroids was increased by treatment with vascular endothelial growth factor (VEGF) or combination of VEGF and fibroblast growth factor-2 (FGF-2). Interestingly, there was no further increase in sprout formation by ECFC+MSC spheroids in response to VEGF or VEGF+FGF-2, suggesting that MSCs stimulate sprout formation by ECFCs. Immuno-fluorescent labeling technique revealed that MSCs surrounded ECFC-mediated sprout structures. We tested vatalanib, VEGF inhibitor, using ECFC and ECFC+MSC spheroids. Vatalanib significantly inhibited sprout formation in both spheroids. Of note, the $IC_{50}$ of vatalanib in ECFC+MSC spheroids at 24 h was $4.0{\pm}0.40{\mu}M$, which are more correlated with the data of previous animal studies when compared with ECFC spheroids ($0.2{\pm}0.03{\mu}M$). These results suggest that ECFC+MSC spheroids generate physiologically relevant sprout structures composed of two types of vascular cells, and will be an effective pre-clinical in vitro assay model to evaluate pro- or anti-angiogenic property.

A Study on Growth of the Green Leaf Lettuce Depends on PPFD and Light Quality of LED Lighting Source for Growing Plant (식물재배용 LED 광원의 광질과 PPFD에 따른 청치마상추의 성장에 관한 연구)

  • Yang, Jun-Hyuk;Choi, Won-Ho;Park, Noh-Joon;Park, Dae-Hee
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.28 no.2
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    • pp.142-147
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    • 2015
  • The artificial light sources for growth of plant are usually high-pressure sodium lamp, metal haloids lamp, and fluorescent light; however, these light sources have relatively weaker Red and Blue lights that are necessary for growth of plants. Especially the effect of Photosynthetic Photon Flux Density (PPFD) is pointed out as the weakness. Meanwhile, LED light source can be selected by specific wavelength to greatly improve the effect of PPFD. In this regard, this paper aims to investigate the promotion of plant growth by measuring photosynthetic photon flux density (hereafter referred to as PPFD) according to changes in light quality of the LED light sources. Towards this end, LED light sources for plant growth were produced with 4 kinds of mono-chromatic lights and 6 kinds of combined lights by mixing red, blue, green and white lights. A comparative analysis was conducted to investigate the effects of optical properties and PPFD on plants (green leaf lettuce) using the produced light sources. The results monochromatic light has fastest growth rate, but plant growth conditions have poor. This being so, mixed light is suitable for the green leaf lettuce.

Therapeutic efficacy of the photoactivated sickle cells as novel drug delivery vehicle (약물전달 시스템 개발을 위한 여기된 광감응제의 응용)

  • Choe, Se-woon
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2015.10a
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    • pp.958-960
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    • 2015
  • Sickle cells possess a unique combination of traits that may enable their use as models for novel synthetic tumor targeting controlled release drug carriers with the ability to treat disseminated tumors in advanced metastatic disease. In this study, we assess the ability of light-activated release sickle cells to enhance tumor delivery of the fluorescent dye calcein by delayed photolysis controlled release compared to free systemic administration of calcein. Sickle cells from mouse models of the disease were shown to preferentially accumulate in tumors compared to adjacent tissue, in 4T1 tumors in mice on a time scale about 12 hours. Sickle cells photosensitized with protoporphyrin IX achieved delayed release of 50% of contents 8-16 hours after photoactivation, which was deemed useful for in vivo delivery of cargo to tumors given the tumor accumulation time of the sickle cells. Sickle cells may be useful as a model for new synthetic drug carrier particles with delayed photolysis controlled release properties.

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Electroluminescent Properties of White Light-Emitting Device Using Photoconductive Polymer and Anthracene Derivatives (광전도성 고분자와 안트라센 유도체를 이용한 백색 전계발광소자의 발광 특성)

  • Lee Jeong-Hwan;Choi Hee-Lack;Lee Bong
    • Korean Journal of Materials Research
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    • v.15 no.8
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    • pp.543-547
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    • 2005
  • Organic electroluminescence devices were made from 1,4-bis-(9-anthrylvinyl)benzene (AVB) and 1,4-bis-(9-aminoanthryl)benzene (AAB) anthracene derivatives. Device structure was ITO/AVB/PANI(EB)/Al (multi-layer device) and ITO/AAB:DCM/Al(single-layer device). In these devices, AVB, polyaniline(emeraldine base) (PANI(EB)) and AAB were used as the emitting material. 4-(dicyanomethylene)-2-methyl-6-p-(dimethylamino)styryl-4H -pyran(DCM) was used as red fluorescent dopant. We studied change of fluorescence wavelength with concentration of DCM doped in AAB. The ionization potential (IP) and optical band gap (Eg) were measured by cyclic voltammetry and UV-visible spectrum. We compared with difference of emitting wavelength between photoluminescence and electroluminescence spectrum. In case of the multi-layer device, PANI and AVB EL spectra have similar wave pattern to each PL spectrum and when PAM and AVB were used at the same time, and multi-layer device showed that a balanced recombination and radiation kom PANI and AVB. In case of the single-layer device, with the increase of DCM concentration, the blue emission decreases and red emission increases. This indicates that DCM was excited by the energy transfer from AAB to DCM or the direct recombination at the dopant sites due to carrier trapping, or both. The device with $1.0wt\%$ DCM concentration gave white light.

Synthesis of Novel Network Polyesters Containing Malonate Group in Main Chain and Their Fluorescence Image Patterning via Photodegradation (주사슬에 말로네이트기를 가지는 신규 폴리에스테르의 합성과 광분해 특성을 이용한 형광 이미지 패터닝)

  • Jeong, Seon-Ju;Kwak, Gi-Seop;Jung, In-Tae;Lee, Dong-Ho;Roh, Hyung-Jin;Yoon, Keun-Byoung
    • Polymer(Korea)
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    • v.32 no.1
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    • pp.56-62
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    • 2008
  • Three types of network polyesters have been newly synthesized by a two-step condensation reaction by the various combination of several diols and diacids. When these polymer films were thermally treated at $240^{\circ}C$, they exhibited absorptions in a visible range despite the forbidden transition of carbonyl group. When excited at wavelengths above 330 nm, the polymers showed fluorescences in a wide visible range from blue to near yellow. These fluorescence phenomena are due to the formation of certain conjugated structures by the Knoevenagel type self-condensation under the high-temperature thermal treatment. These polymers showed significant difference in the thermal properties as a function of the degrees of chemical crosslinking. They also underwent photodegradation. Highly resolved, fluorescent image patterns were successfully obtained by the photodegradation of malonate group under a strong UV-light irradiation.