• Title/Summary/Keyword: Confocal laser scanning microscopy

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Fluorescent Silk Fibroin Nanoparticles Prepared Using a Reverse Microemulsion

  • Myung, Seung-Jun;Kim, Hun-Sik;Kim, Yeseul;Chen, Peng;Jin, Hyoung-Joon
    • Macromolecular Research
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    • v.16 no.7
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    • pp.604-608
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    • 2008
  • Color dye-doped silk fibroin nanoparticles were successfully fabricated using a microemulsion method. An aqueous silk fibroin solution was prepared by dissolving cocoons (Bombyx mori) in a concentrated lithium bromide solution followed by dialysis. A color dye solution was also mixed with the aqueous silk fibroin solution. The surfactants used for the microemulsion were then removed by methanol and ethanol, yielding color dye-doped silk fibroin nanoparticles, approximately 167 nm in diameter. The secondary structure of the nanoparticles showed a $\beta$-sheet conformation, as characterized by Fourier transform infrared spectroscopy. The morphology of the nanoparticles was determined by field emission scanning electron microscopy, transmission electron microscopy and atomic force microscopy, and their size and size distribution were measured by dynamic light scattering. The color dye-doped silk fibroin nanoparticles were examined by confocal laser scanning microscopy.

Carbide Behavior in STD11 Tool Steel during Heat Treatment (STD11 공구강의 열처리 온도에 따른 탄화물 거동)

  • Hong, Ki-Jung;Song, Jin-Hwa;Chung, In-Sang
    • Journal of the Korean Society for Heat Treatment
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    • v.24 no.5
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    • pp.262-270
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    • 2011
  • Carbide precipitation and dissolution behavior at various temperatures during heat treatment has been studied in STD11 cold working die steel through confocal scanning laser microscopy; dilatometry; and X-ray diffraction analysis. The equilibrium phase diagram and phase fractions with temperature were calculated using a FactSage program. Confocal laser microscopic observation revealed that ${\alpha}$ to ${\gamma}$ transformation temperature is near $800^{\circ}C$; M7C3 carbides melt at $1245^{\circ}C$; and the melting temperature of STD11 is near $1370^{\circ}C$. XRD results indicated that the M23C6 carbides dissolve in the matrix if austenitized at over $1030^{\circ}C$; while the M7C3 carbides remain up to $1200^{\circ}C$ although their amount decreases. The calculated equilibrium phase diagram showed good agreement with experimental results on carbide dissolution and phase transformation temperatures.

Droplet Geometry and Its Volume Analysis (기름방울 형상 및 그 체적 분석법)

  • Yoon, Moon-Chul
    • 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.

Effect of three different irrigation solutions applied by passive ultrasonic irrigation

  • Llena, Carmen;Forner, Leopoldo;Cambralla, Raquel;Lozano, Adrian
    • 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.

The bactericidal effect of an atmospheric-pressure plasma jet on Porphyromonas gingivalis biofilms on sandblasted and acid-etched titanium discs

  • Lee, Ji-Yoon;Kim, Kyoung-Hwa;Park, Shin-Young;Yoon, Sung-Young;Kim, Gon-Ho;Lee, Yong-Moo;Rhyu, In-Chul;Seol, Yang-Jo
    • Journal of Periodontal and Implant Science
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    • v.49 no.5
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    • pp.319-329
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    • 2019
  • Purpose: Direct application of atmospheric-pressure plasma jets (APPJs) has been established as an effective method of microbial decontamination. This study aimed to investigate the bactericidal effect of direct application of an APPJ using helium gas (He-APPJ) on Porphyromonas gingivalis biofilms on sandblasted and acid-etched (SLA) titanium discs. Methods: On the SLA discs covered by P. gingivalis biofilms, an APPJ with helium (He) as a discharge gas was applied at 3 different time intervals (0, 3, and 5 minutes). To evaluate the effect of the plasma itself, the He gas-only group was used as the control group. The bactericidal effect of the He-APPJ was determined by the number of colony-forming units. Bacterial viability was observed by confocal laser scanning microscopy (CLSM), and bacterial morphology was examined by scanning electron microscopy (SEM). Results: As the plasma treatment time increased, the amount of P. gingivalis decreased, and the difference was statistically significant. In the SEM images, compared to the control group, the bacterial biofilm structure on SLA discs treated by the He-APPJ for more than 3 minutes was destroyed. In addition, the CLSM images showed consistent results. Even in sites distant from the area of direct He-APPJ exposure, decontamination effects were observed in both SEM and CLSM images. Conclusions: He-APPJ application was effective in removing P. gingivalis biofilm on SLA titanium discs in an in vitro experiment.

Biomineralization of Calcium Carbonate Polymorphs by the Bacterial Strains Isolated from Calcareous Sites

  • Dhami, Navdeep Kaur;Reddy, M. Sudhakara;Mukherjee, Abhijit
    • Journal of Microbiology and Biotechnology
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    • v.23 no.5
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    • pp.707-714
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    • 2013
  • Microbially induced calcium carbonate precipitation (MICCP) is a naturally occurring biological process that has various applications in remediation and restoration of a range of building materials. In the present investigation, five ureolytic bacterial isolates capable of inducing calcium carbonate precipitation were isolated from calcareous soils on the basis of production of urease, carbonic anhydrase, extrapolymeric substances, and biofilm. Bacterial isolates were identified as Bacillus megaterium, B. cereus, B. thuringiensis, B. subtilis, and Lysinibacillus fusiformis based on 16S rRNA analysis. The calcium carbonate polymorphs produced by various bacterial isolates were analyzed by scanning electron microscopy, confocal laser scanning microscopy, X ray diffraction, and Fourier transmission infra red spectroscopy. A strain-specific precipitation of calcium carbonate forms was observed from different bacterial isolates. Based on the type of polymorph precipitated, the technology of MICCP can be applied for remediation of various building materials.

Generation and characterization of a monoclonal antibody with high species-specificity to Schistosoma japonicum glutathione S-transferase

  • Kim, Jung-Hwan;Park, Jung-Hyun;Ju, Sung-Kyu;Lee, Myung-Kyu;Kim, Kil Lyong
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.187-195
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    • 2001
  • The expression of recombinant proteins fused to 26 kDa glutathione S-transferase (GST) extracted from Schistosoma japonicum represents an attractive system for purifiying proteins of interest in a single step using GST-affinity chromatography. In addition, the GST-tag is used conveniently for detecting fused proteins since its high solubility as well as its relatively small size rarely interferes with the biological activity of the fused protein. In this regard, the GST system is frequently applied for tracing fusion proteins in both prokaryotic and eukaryotic cells to elucidate the physiological interactions and functional compartments of proteins. To provide a further tool in analyzing GST-fusion proteins, a new monoclonal antibody, with a high specificity to the S. japonicum GST was produced. Methods: BALB/c mice were immunized both with recombinant S. japonicum GST proteins, and by the fusion of splenocytes from these mice with myeloma cells. From this, a new anti -GST monoclonal antibody, termed SARAH, was generated. The specificity and reactivity of this antibody was confirmed by ELISA and by Western blot analysis. Results: SARAH showed a high reactivity to recombinant GST and GST fusion protein but not with native mammalian GST proteins as derived from other species including humans, cows, rabbits and rats. The applicability of SARAH was further demonstrated by confocal laser scanning microscopy, where GST proteins that were expressed transiently in mouse fibroblast cells, were specifically detected without interference of endogenous GST. Conclusion: SARAH is new monoclonal antibody with a high specificity to recombinant GST proteins but not to endogenous GST in mammalian cells.

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Antimicrobial Effect of Photodynamic Therapy Using Plaque Disclosing Agent (치면착색제를 이용한 광역동 치료의 항균 효과)

  • Kho, Junhee;Park, Howon;Lee, Juhyun;Seo, Hyunwoo;Lee, Siyoung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.47 no.2
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    • pp.120-127
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    • 2020
  • The aim of this study was to evaluate the antimicrobial effect of photodynamic therapy (PDT) using plaque disclosing agent, 10 - 20 mM erythrosine, as a photosensitizer. Multispecies cariogenic biofilms containing Streptococcus mutans, Lactobacillus casei and Candida albicans were formed on hydroxyapatite disc. 20 μM, 10 mM and 20 mM erythrosine were applied as a photosensitizer for 3 minutes, and then light-emitting diode (LED) irradiated for 24 seconds. Colony-forming unit (CFU) were measured and biofilms were observed using confocal laser scanning microscopy (CLSM). CFU were significantly decreased in the PDT groups using 10 - 20 mM erythrosine (10 mM, 20mM) and the results were also confirmed by CLSM. This study confirms the high antimicrobial effect of photodynamic therapy using plaque disclosing agent as a photosensitizer.