• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.703-706
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• 2000

We constructed a biospecific affinity surface using hyper-branched dendrimers on gold for biospecific recognition, and characterized the resulting surfaces by using confocal fluorescence microscopy. The dendrimer monolayer was firstly constructed on the mercaptoundecanoic acid SAM/Au with pentafluorophenyl ester activation and further functionalized with sulfo-NHS-biotin, an activated ester of biotin. To confirm the formation of biospecific affinity surface, FITC(fluorescein isothiocyanate)-labeled avidin was loaded onto the biotinylated dendrimer monolayer, and fluorescence images of the bound avidins were investigated with a confocal microscope. The constructed biospecific affinity surface showed a much more dense and uniform fluorescence compared to those from poly-L-lysine- and cystamine SAM-based affinity surfaces. For the dependency on the concentration of added FITC-labeled avidin on the affinity surface, derived fluorescence could be detectable from as low as $1{\mu}g/ml$, and intensified up to $50{\mu}g/ml$. Further reaction of FITC-labeled avidin layer with TMR(tetramethylrhodamine)-biocytins resulted in the efficient FRET(fluorescence resonance energy transfer) phenomenon. As an extension of the study, we attempted a patterning of the affinity surfaces on gold by microcontact printing. Fluorescence of the patterned surface demonstrated that FITC-labeled avidin molecules were specifically bound to the biotinylated patches.

• Tribology and Lubricants
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• v.24 no.6
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• pp.320-325
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• 2008

The recent industrial application requires technical methods to get the cutting fluid droplet surfaces in particular from the viewpoint of topography and micro texture. To characterize the surface topography of droplet, the combination of the confocal laser scanning microscope (CLSM) and wavelet filtering is well suited for obtaining the droplet geometry encountered in tribological research. This technique indicates a better agreement in obtaining an appropriate droplet surface obtained by the CLSM over a detail range of surface accuracy (resolution: $2{\mu}m$). And the results allow an excellent accuracy in a measurement of a droplet surface. The combination of extended focal depth measurement configured and multi-scale wavelet filtering has proven that it can construct a droplet surface in a successive and accurate way. A multi-scale approach of wavelet filtering was developed based on the decomposition and reconstruction of droplet surface by 2D wavelet transform using db9 (a mother wavelet of daubechies). Also this technique can be extended to characterize the quantification of droplet properties and other field in a wide range of scales. Finally this method is verified to be a better droplet surface modeling in a micro scale arising in a mist machining.

• Biomolecules & Therapeutics
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• v.26 no.2
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• pp.182-190
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• 2018

The objective of this work is to evaluate the effect of polyamidoamine (PAMAM) dendrimers on electroosmotic flow (EOF) through skin. The effect of size and concentration of dendrimer was studied, using generation 1, 4 and 7 dendrimer (G1, G4 and G7, respectively). As a marker molecule for the direction and magnitude of EOF, a neutral molecule, acetoaminophen (AAP) was used. The visualization of dendrimer permeation into the current conducting pore (CCP) of skin was made using G4-fluorescein isothiocyanate (FITC) conjugate and confocal microscopy. Without dendrimer, anodal flux of AAP was much higher than cathodal or passive flux. When G1 dendrimer was added, anodal flux decreased, presumably due to the decrease in EOF by the association of G1 dendrimer with net negative charge in CCP. As the generation increased, larger decrease in anodal flux was observed, and the direction of EOF was reversed. Small amount of methanol used for the preparation of dendrimer solution also contributed to the decrease in anodal flux of AAP. Cross-sectional view perpendicular to the skin surface by confocal laser scanning microscope (CLSM) study showed that G4 dendrimer-FITC conjugate (G4-FITC) can penetrate into the viable epidermis and dermis under anodal current. The permeation route seemed to be localized on hair follicle region. These results suggest that PAMAM dendrimers can permeate into CCP and change the magnitude and direction of EOF. Overall, we obtained a better understanding on the mechanistic insights into the electroosmosis phenomena and its role on flux during iontophoresis.

• Restorative Dentistry and Endodontics
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• v.40 no.2
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• pp.143-148
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• 2015

Objectives: This study evaluated the maximum depth and percentage of irrigant penetration into dentinal tubules by passive ultrasonic irrigation (PUI). Materials and Methods: Thirty extracted human teeth were instrumented and divided into three groups. According to final irrigation regimen, 5.25% sodium hypochlorite (Group A, NaOCl), 2% chlorhexidine (Group B, CHX) and saline solution (Group C, control group) were applied with Irrisafe 20 tips (Acteon) and PUI. Irrigant was mixed with 0.1% rhodamine B. Sections at 2 mm, 5 mm, and 8 mm from the apex were examined with confocal laser scanning microscopy (CLSM). The percentage and maximum depth of irrigant penetration were measured. Kruskal-Wallis test and Mann-Whitney test were performed for overall comparison between groups at each level and for pairwise comparison, respectively. Within a group, Wilcoxon test was performed among different levels. p values less than 0.05 were considered significant. Results: In all groups, highest penetration depth and percentage of penetration were observed at the 8 mm level. At 2 mm level, Groups A and B had significantly greater depths and percentages in penetration than Group C (p < 0.05), but there were no significant differences between Groups A and B. At 5 mm level, penetration depths and percentage of penetration was not significantly different among the groups. Conclusions: NaOCl and CHX applied by PUI showed similar depth and percentage of penetration at all evaluated levels.

• Korean Journal of Optics and Photonics
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• v.15 no.6
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• pp.583-590
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• 2004

The autofocus is one of the important processes in the automated vision inspection or measurements using optical microscopes, because it influences the measuring accuracy. In this paper, we used the confocal microscope configuration based on not a pinhole but a single-mode optical fiber. A single mode fiber has the functions of source and detector by applying the reciprocal scheme. As a result, we acquired a simple system configuration and easy alignment of the optical axis. Also, we embodied a fast autofocus system by acquiring the focus error signal through a source modulation technique. The source modulation technique can effectively reduce physical disturbances compared with objective lens modulation, and it is easily applicable to general optical microscopes. The focus error signal was measured with respect to the modulation amplitude, reflectance of the specimen and inclination angle of the measuring surface. The performance of the proposed autofocus system was verified through autofocusing flat mirror surface. In addition, we confirmed that source modulation rarely degrades the depth resolution by the comparison between the FWHMs of axial response curves.

• Restorative Dentistry and Endodontics
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• v.27 no.3
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• pp.257-269
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• 2002

Objective : The purpose of this study was to evaluate the resin infiltration into dentin of one-bottle adhesive systems and self-etching primer bonded to Class V cavities using confocal laser scanning microscope(CLSM). Material and Methods : Forty Class V cavities were prepared from freshly extracted caries-free Human teeth. These teeth were divided into two groups based on the presence of cervical abrasion: Group I, cervical abrasion : Group II, wedge-shaped cavity preparation. Resin-dentin interfaces were produced with two one-bottle dentin bonding systems-ONE COAT BOND(OCB; Coltene$^R$) and Syntac$^R$SPrint$^{TM}$(SS; VIVADENT)-, one self-etching priming system-CLEARFIL$^{TM}$ SE BOND (SB : KURARAY)- and one multi-step dentin bonding system-Scotchbond$^{TM}$Multi-Purpose (SBMP, 3M Dental Products)-as control according to manufacturers' instructions. Cavities were restored with Spectrum$^{R}$(Dentsply). Specimens were immersed in saline for 24 hours and sectioned longitudinally with a low-speed diamond disc. The resin-dentin interfaces were microscopically observed using CLSM. The quality of resin-infiltrated dentin layers were evaluated by five dentists using 0~4 scale. Results : Confocal laser scanning microscopal investigations using primer labeled with rhodamine B showed that the penetration of the primer occurred along the cavity margins. Statistical analysis using one-way ANOVA followed by Duncan's Multiple Range test revealed that the primer penetration of the group 2(wedge-shaped cavity preparation) was more effective than group 1(cervical abrasion) and that of the gingival interfaces was more effective than the occlusal interfaces. In the one-bottle dentin bonding systems, the resin penetration score of OCB was compatible to SBMP, but those of SS and self-etching priming system, SB were lower than SBMP.

• Journal of the korean academy of Pediatric Dentistry
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• v.45 no.2
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• pp.137-143
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• 2018

The aim of this study is to compare the differences of the demineralization resistance of resin infiltration and 1.23% acidulated phosphate fluoride in bovine teeth with artificial caries. We applied 1.23% Acidulated phosphate fluoride (APF) gel and $Icon^{(R)}$ caries infiltrant on the artificial bovine enamel carious lesion and then demineralized all samples. The depth of demineralization was measured by using Confocal Laser Scanning Microscope (CLSM) and observed the roughness and irregularity of the enamel was observed by Scanning Electron Microscope (SEM). In this experiment with demineralization resistance on smooth artificial carious lesion, less depth of demineralization, roughness, and irregularity of enamel was observed in APF gel and $Icon^{(R)}$ group than in the control group. There was no significant difference between the depth of demineralization of 1.23% APF gel and $Icon^{(R)}$ caries infiltrant group. However, resin infiltration is beneficial as less roughness and irregularity was observed on the enamel surface than when 1.23% APF gel is applied.

• The Journal of Korean Medicine
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• v.27 no.3 s.67
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• pp.107-131
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• 2006

Background and Aims: Dansamtongmek-tang (DSTMT) and Dansamsengmek-san (DSSMS) have been used for many years as therapeutic agents for the acute stage of cerebrovascular disease, hypertension and hyperlipidemia in Oriental medicine, but the effects of DSTMT and DSSMS on hyperlipidemia and safety for cell damage are not yet well-known. This study was done to investigate the effects of DSTMT and DSSMS on hyperlipidemia. Methods: In vivo test: after administering DSTMT and DSSMS to SHR and ICR occurred hyperlipidemia for 3 weeks, we analyzed body weight, cholesterol levels. TG, HDL-chol, LDL-chol, LDH in plasma, brain, liver and kidney tissue, and DNA by RT-PCR. In vitro test: after administering DSTMT and DSSMS to human hepatocellular carcinoma in hypoxia, we observed cell cohesion by light microscope, analyzed the inflow of Ca2+ by confocal laser scanning microscope and DNA by RT-PCR. Results: DSTMT significantly decreased the levels of triglyceride and increased the levels of HDL-cholesterol in SHR, and significantly decreased the levels of LDL-cholesterol and body weight and increased the levels of HDL-cholesterol in ICR. DSSMS significantly decreased body weight, total cholesterol levels, LDL-cholesterol, LDH and cardiac risk factor (CRE) in SHR and significantly decreased the levels of total cholesterol, triglyceride, LDL-cholesterol, LDH and CRF in ICR. DSTMT had an effect on protecting cells from damage by inhibiting production of p53 mRNA, and in DSSMS, by inhibiting production of p53 mRNA and p21 mRNA after hypoxia. DSTMT effectively blocked off Ca2+ at low density, but DSSMS effectively blocked off Ca2+ at high density. Both DSTMT and DSSMS had an effect on inhibiting lipid metabolism by blocking off production of apo B mRNA. Conclusions: These results suggest that DSTMT and DSSMS might be usefully applied for treatment of hyperlipidemia and suppression of brain damage.

• Restorative Dentistry and Endodontics
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• v.25 no.4
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• pp.575-586
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• 2000

In this study, adaptation of compomer to saliva contaminated dentin was evaluated with scanning electron microscope(SEM) and confocal laser scanning microscope(CLSM). For the SEM study, the occulusal surfaces of thirty two molar teeth were grounded to exposure dentin surfaces. The specimen were randomly assigned to control and three experimental groups with four samples in each group. In control group, Dyract and F-2000 compomer were bonded on the specimens according to the manufactures direction. Experimental groups were subdivided into three groups. They were contaminated with saliva on dentin surfaces ; Experimental group 1 : Saliva was dried with compressed air. Experimental group 2 : Saliva was rinsed with air-water spray and dried. Experimental group 3 : After polymerization of an adhesive, they were contaminated with saliva, and then saliva was rinsed with air-water spray and dried. Dyract and F-2000 compomer were bonded on saliva-treated dentin surfaces. The interfaces between dentin and compomer were observed with SEM. For the CLSM study, Class V cavities were prepared in buccal and ligual surfacess of thirty two molars. The specimens were divided into control and experimental groups. Class V cavities in experimental group were contaminated with saliva and those surfaces in each experimental groups received the same treatments as for the SEM study. Cavities were applied Prime & Bond 2.1 and F-2000 compomer primer/adhesive that were mixed with fluorescein, and then were filled with Dyract and F-2000 compomer. Specimens were embedded in transparent acrylic resin and sectioned buccolingual1y with diamond wheel saw, and then mounted on cover slide for CLSM study. The interface between cavity and compomer was observed by fluoresence imaging with a CLSM. The results were as follows : 1. In SEM exammination of Dyract group, control group, experimental group 2, 3 showed close adaptation to dentin and hybrid layer of $3{\sim}4{\mu}m$ diameter. Interfacial gap between compomer and dentin in experimental group 1 was wider than in control group. 2. In SEM examination of F-2000 group, adaptation to dentin of control group was closer than Dytact control group, but hybrid-like layer was not observed. Interfacial gap between compomer and dentin in experimental group 1 was wider than in Dyract experimental group 1. 3. In dissolution specimens of Dyract and F-2000 group, resin tags penetrated through dentinal tubules in control group and experimental group 1 and 3, but the penetration of resin tag was irregular and partial in experimental group 1. 4. In CLSM exammination of Dyract and F-2000 group, adhesive patterns of control and experimental groups showed same as in SEM. This result suggests the treatment methods, rinsing & drying, repeating all adhesive procedures, will produce good effect on adaptation of compomer to dentin if the dentin surface or polymerized adhesive is contaminated by saliva.

• Development and Reproduction
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• v.1 no.1
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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.