• 대한기계학회논문집A
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• 제28권9호
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• pp.1429-1434
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• 2004

Immunoassay is one of the important analytical methods for clinical diagnoses and biochemical studies, but needs a long time, troublesome procedures and expensive reagents. In this study, therefore, we propose the micro filter chip with microbeads for immunoassay, which has pillar structures. The advantage of the proposed micro filter chip is to use simple fabrication process and cheap materials. The mold was made by the photolithography technique with Si wafer and negative photoresist SU-8. The replica was made of PDMS, bonded on the pyrex glass. The micro filter chip consists of inlet channel, filter chamber and outlet channel. HBV (Hepatitius B virus) monoclonal antibody (Ag1) labeled with biotin were immobilized onto streptavidin coated beads of 30∼50 $\mu$m size. Fluorescein isothiocyanate (FITC)-labeled HBV monoclonal antibody (Ag8) was used to detect HBsAg (Hebatitis B virus surface Antigen), and fluorescence intensity was monitored by epi-fluorescence microscope. In this study, the immune response of less than 30 min was obtained with with the use of 100 $m\ell$ of sample.

• Journal of Photoscience
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• 제12권3호
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• pp.123-127
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• 2005

The polyclonal antibody was generated against the peptide fragment of 62 amino acid residues (D 181-T242) near the COOH-terminal region of the extracellular domain of epithelial-cell adhesion molecule (Ep-CAM) and shown to be able to recognize Ep-CAM in competitive ELISA. Then, sulforhodamine 101 acid chloride (so called Texas red), a fluorescence dye, was conjugated to the affinity-purified anti-Ep-CAM antibody utilizing the reaction between the aliphatic amines of antibody and the sulfonyl chloride of Texas red. The molar ratio of Texas red to antibody was estimated to be approximately 1.86 by measuring optical densities at 280 nm and 596 nm, implying that the two molecules of Texas red at most were conjugated to antibody. The anti-Ep-CAM antibody-Texas red conjugate was then used for immunohistochemistry of CT-26 murine colon carcinoma cells. Based upon the fluorescence microscope images, anti-Ep-CAM antibody is able to deliver Texas red specifically to the surface of CT-26 cells on which Ep-CAM was actively expressed. This result indicates that anti-Ep-CAM antibody could be useful for the tissue-specific delivery of photosensitizers via antigen-antibody interaction.

• KSBB Journal
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• 제28권3호
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• pp.191-195
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• 2013

Cordyceps militaris (CM) occurring from a fruiting body by a host insect is a kind of mushroom, which is composed of animal host and plant fruit body. CM contains large amounts of useful ingredients including polysaccharides, vitamins, amino acids, minerals, etc. The essential amino acids from CM including cystine, lysine, and methionine can be expected to improve damaged hair treatment as effective ingredients. In this study, the improvement effect of the CM extracts on chemical and physical properties for damaged-hair treatment was investigated. The cysteic acid and cystine monooxide produced by oxidation of cystine were analyzed their chemical structure by FT-IR spectroscopy. It was confirmed that the vibration absorption peak ($1,041cm^{-1}$) of cysteic acid was reduced and increased sulfur content considerably which means meaningful improvement effect on damaged-hair treatment. Apparently, the cuticle morphology of the damaged-hair was improved significantly by treatment with CM extracts. Especially, confocal laser scanning microscope images of the damaged-hair treated with the extract showed highly increased fluorescence intensity which means promising effect in hair treatment. The tensile strength of the damaged hair treated also increased by 168% compared with the damaged hair.

• Bulletin of the Korean Chemical Society
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• 제30권7호
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• pp.1521-1525
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• 2009

Thermosensitive block copolymers of ethylene oxide and N-isopropylacrylamide (NIPAM) were synthesized. A five armed star shape oligo(ethylene oxide) initiator with a cyclotriphosphazene core was prepared and used for the atom transfer radical polymerization (ATRP) of NIPAM. The lower critical solution temperatures (LCSTs) of the copolymers were 36 to 46 ${^{\circ}C}$, higher than that of PNIPAM (32 ${^{\circ}C}$), depending on their molecular weights. The copolymers were soluble in water below the LCSTs but formed micelles above the LCSTs. The thermosensitive micellization behaviors of the polymers were investigated by fluorescence spectroscopy. With increasing the temperature of an aqueous solution of P2 and pyrene above the LCST, the peak of 333 nm red-shifted to appear around 339 nm and its intensity increased significantly, indicating the micelle formation. The transfer of pyrene into the micelles was also confirmed by a confocal laser scanning microscope. The fluorescence image obtained from P2 in an aqueous pyrene solution exhibited a green emission resulting from the pyrene transferred into the micelles. Salt effects on the solubility of the copolymers in an aqueous solution were investigated. The LCST of P2 decreased sharply as the concentration of sodium chloride increased, while decreased slowly with potassium chloride.

• 약학회지
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• 제49권2호
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• pp.174-179
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• 2005

The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmen tal facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Campsis grandiflora extract on the UVB-irradiated human dermal fibroblasts (HDFs). We tested free radical and superoxide scavenging effect in vitro. C. grandiflora extracts had potent radical scavenging effect by 82% at $100{\mu}g/ml$, respectively. For testing intracellular ROS scavenging activity the cultured HDFs were analyzed by increase in DCF fluorescence upon exposure to UVB 20 $MJ/cm^2$ after treatment of C.grandiflora extracts. The results showed that oxidation of CM-DCFDA was inhibited by C.grandiflora extracts effectively and C.grandiflora extracts has a potent free radical scavenging activity in UVB- irradiated HDFs. In ROS imaging using confocal microscope we visualized DCF fluorescence in HDFs directly. In conclusion, our results suggest that C.grandiflora can be effectively used for the prevention of UV-induced adverse skin reactions such as radical production, and skin cell damage.

• Restorative Dentistry and Endodontics
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• 제26권1호
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• pp.1-8
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• 2001

Dental Caries which has high prevalence rate, accounts for majority of dental diseases. Many treatment and preventive treatment has been developed, thereby reducing the prevalence rate, but in our country, fluoridization has not spread widely yet, so prevention has not been done satisfactorily. When dental caries progresses, irreversible damage of tooth structure occurs. In initial dental caries, demineralizing tooth structure can be remineralized, so restorative treatment is unnecessary. In this study, 20 teeth restored with composite resin without fluoride release were used and divided into two groups. Incipient dental caries were artificially made and demineralization procedure was done for 1 and 2 weeks, for each group. Changes in mineral contents around the margins were analysed with confocal laser scanning microscope. The results were as follow. 1. Both total fluorescence of the lesion and average fluorescence of the lesion of remineralized samples decreased compared to demineralizing state. (p＜0.01) 2. Confocal laser scanning microscopy can be used in quantitative analysis of mineral change. In result, confocal laser scanning microscopy can be used in quantitative analysis of mineral change and it could be used in many different fields of dentistry in the future.

• Journal of Plant Biotechnology
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• 제48권3호
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• pp.156-164
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• 2021

Spirodela polyrhiza, from the Lemnaceae family, are small aquatic plants that offer an alternative plant-based system for the expression of recombinant proteins. However, no turion transformation protocol has been established in this species. In this study, we exploited a pB7YWG2 vector harboring the eYFP gene that encodes enhanced yellow fluorescent protein (eYFP), which has been extensively used as a reporter and marker to visualize recombinant protein localization in plants. We adopted Agrobacterium tumefaciens-mediated turion transformation via vacuum infiltration to deliver the eYFP gene to turions, special vegetative forms produced by duckweeds to endure harsh conditions. Transgenic turions regenerated several duckweed fronds that exhibited yellow fluorescent emissions under a fluorescence microscope. Western blotting verified the expression of the eYFP protein. To the best of our knowledge, this is the first report of an efficient protocol for generating transgenic S. polyrhiza expressing eYFP via Agrobacterium tumefaciens-mediated turion transformation. The ability of turions to withstand harsh conditions increases the portability and versatility of transgenic duckweeds, favoring their use in the further development of therapeutic compounds in plants.

• 대한바이러스학회지
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• 제28권3호
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• pp.225-232
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• 1998

We have expressed GFP in Sf9 and Bm5 cells or Bombyx mori larvae by using Ac-Bm hybrid virus capable of replicating in both Bm5 and Sf9 cells. Genomic DNA of Ac-Bm hybrid virus expressing ${\beta}$-galactosidase was cotransfected with baculovirus transfer vector containing GFP gene, pBacPAK-GFP in Sf9 cells. The Ac-Bm hybrid virus harboring GFP was named as Ac-Bm hybrid virus-GFP. The Ac-Bm hybrid virus-GFP-infected insect cells were easily selected by detecting the emission of GFP from each well of cell culture dish on the UV illuminator. GFP produced by Ac-Bm hybrid virus-GFP in Sf9 and Bm5 cells or B. mori larvae was confirmed by SDS-PAGE and Western blot analysis using GFP antibody. In addition, B. mori larvae infected with Ac-Bm hybrid virus-GFP was apparently appeared fluorescence from the whole body at S days postinoculation. The fluorescence of GFP from the hemolymph and fat body of B. mori larvae infected with Ac-Bm hybrid virus-GFP was also observed by fluorescence microscope. In conclusion, our results demonstrated that in baculovirus expression vector system, use of Ac-Bm hybrid virus have an additional advantage of expanded host range for producing recombinant proteins.

• 한국발생생물학회지:발생과생식
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• 제1권1호
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• pp.45-56
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• 1997

Befor fertilization, mammalian oocytes undergo meiotic maturation, which consists of nuclear and cytoplasmic differentiation. In this study, changes of $Ca^{2+}$ stores in mouse oocytes were examined during meiotic maturation and the role of $Ca^{2+}$ in the regulation of the maturation was investigated by using monoclonal antibodies against smooth endoplasmic reticulum $Ca^{2+}$-ATPase(SERCA-ATPase) and calreticulin. Observations were made under epifluorescence microscope and/or confocal laser scanning microscope. In immature oocytes which did not resume meiotic maturation, SERCA-ATPases were mostly localized in the vicinity of the germinal vesicle and calreticulins were distributed evenly throughout the cytoplasm. In mature oocytes, SERCA-ATPases were observed throughout the cytoplasm, butwere absent from the nuclear region. In contrast, calreticulins were localized mostl in the cortex of the oocyte and were absent from the cytoplasm. However, bright fluoresence stainings were wbserved in the perimeiotic spindle region of mature oocyte when labeled with antibodies against calreticulin. These results indicate that mouse oocytes undergo distinct rearrangement of the localization of $Ca^{2+}$-ATPases and calreticulins during meiotic maturation. Thus it can be suggested that redistribution of the $Ca^{2+}$ stores, as revealed by differential fluorescence stainings, is deeply involved in the regulatory mechanism of mammalian oocyte maturation.

• 한국주조공학회지
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• 제18권6호
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• pp.550-554
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• 1998

The microstructures of rapidly solidified binary Ti-Al alloys containing $45{\sim}58\;at%Al$ have been studied using C/S (carbon/sulfur), N/O (nitrogen/oxygen) analyser, X-ray fluorescence spectrometer (XRF), X-ray diffractometer (XRD), optical microscope (OM) and scanning electron microscope (SEM). The phases present in the alloys and their distribution were found to be a sensitive function of Al content. Essentially single-phase (${\gamma}$) microstructures were observed to alloys with 45 at%Al, 55 at%Al and 58 at%Al. In other content alloys, two phase (${\alpha}_2$, ${\gamma}$) microstructures were observed. The 48 at%Al, 52 at%Al alloys contain (${\gamma}+{\alpha}_2$) phase and ${\alpha}_2$ phase. These results indicate that rapid solidification affect the solidification path, then metastable phase forming during solidification.