• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1209-1215
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• 2017

Microwave sterilization was performed to inactivate the spores of biofilms of Bacillus cereus involved in foodborne illness. The sterilization conditions, such as the amount of water and the operating temperature and treatment time, were optimized using statistical analysis based on 15 runs of experimental results designed by the Box-Behnken method. Statistical analysis showed that the optimal conditions for the inactivation of B. cereus biofilms were 14 ml of water, $108^{\circ}C$ of temperature, and 15 min of treatment time. Interestingly, response surface plots showed that the amount of water is the most important factor for microwave sterilization under the present conditions. Complete inactivation by microwaves was achieved in 5 min, and the inactivation efficiency by microwave was obviously higher than that by conventional steam autoclave. Finally, confocal laser scanning microscopy images showed that the principal effect of microwave treatment was cell membrane disruption. Thus, this study can contribute to the development of a process to control food-associated pathogens.

• KSBB Journal
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• 제26권5호
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• pp.422-426
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• 2011

Alginate bead has been supplemented with various polymers to control permeability and to enhance mechanical strength. In this report, fibroin-reinforced alginate hydrogel was prepared, in which spatial localization of fibroin molecules was investigated. Confocal laser scanning microscopy revealed that fibroin molecules formed a fibrous network in the alginate-fibroin beads, which was expected to enhance mechanical strength as same as in many composite materials. Uniaxial compression test showed that fibroin-reinforced alginate beads had increased mechanical strength only after methanol treatment that caused ${\beta}$-sheet formation among fibroin molecules. Simultaneous curing and dialysis of alginate beads were carried out to remove excesscalcium but to retain fibroin in the dialysis chamber, which fabricated beads without internal fibrous fluorescent stains. Fibroin molecules were only found beneath the surface of the beads. The fibroin-diffused shell was further processed to form a thick wall after drying or was mobilizedto the centre of the bead by methanol treatment. Accordingly, the structure analyses provide processing methods of fibroin to form a wall or center clumps, which could be applied to design controlled delivery device.

• Journal of Pharmaceutical Investigation
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• 제34권2호
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• pp.101-106
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• 2004

This work aims at examining the cellular uptake behavior of poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles derivatized with a protein transduction domain (PTD) using HeLa cells. For this purpose, $Tat_{49-57}$ peptide derived from transcriptional activation (Tat) protein of HIV type-1 was covalently conjugated to the terminal end of PLGA. Nanoparticles were ten prepared with the $Tat_{49-57}-PLGA$ conjugates by a spontaneous phase inversion method. The prepared particles had a mean diameter of ca. 84 nm, as measured by dynamic light scattering. The interaction of the Tat-PLGA nanoparticles with cells was examined by using confocal laser scanning microscopy. It was found tat Tat-PLGA nanoparticles incubated with HeLa cells could efficiently translocate into cytoplasm, while plain PLGA nanoparticles showed negligible cellular uptake. In addition, even at $4^{\circ}C$ or in the presence of sodium azide significant cellular internalization of Tat-PLGA nanoparticles was still observed. These results indicate that a non-endocytotic translocation mechanism might be involved in the cellular uptake of Tat-PLGA nanoparticles.

• Journal of Microbiology and Biotechnology
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• 제18권5호
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• pp.946-951
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• 2008

The opd gene, encoding organophosphorus hydrolase (OPH) from Flavobacterium sp. capable of degrading a wide range of organophosphate pesticides, was surface- and intracellular-expressed in Synechococcus PCC7942, a prime example of photoautotrophic cyanobacteria. OPH was displayed on the cyanobacterial cell surface using the truncated ice nucleation protein as an anchoring motif. A minor fraction of OPH was displayed onto the outermost surface of cyanobacterial cells, as verified by immunostaining visualized under confocal laser scanning microscopy and OPH activity analysis; however, a substantial fraction of OPH was buried in the cell wall, as demonstrated by proteinase K and lysozyme treatments. The cyanobacterial outer membrane acts as a substrate (paraoxon) diffusion barrier affecting whole-cell biodegradation efficiency. After freeze-thaw treatment, permeabilized whole cells with intracellular-expressed OPH exhibited 14-fold higher bioconversion efficiency ($V_{max}/K_m$) than that of cells with surface-expressed OPH. As cyanobacteria have simple growth requirements and are inexpensive to maintain, expression of OPH in cyanobacteria may lead to the development of a low-cost and low-maintenance biocatalyst that is useful for detoxification of organophosphate pesticides.

• 한국멀티미디어학회논문지
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• 제11권12호
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• pp.1635-1648
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• 2008

The extraction of important features in cancer cell image analysis is a key process in grading renal cell carcinoma. In this study, we applied three-dimensional (3D) texture feature extraction methods to cell nuclei images and evaluated the validity of them for computerized cell nuclei grading. Individual images of 2,423 cell nuclei were extracted from 80 renal cell carcinomas (RCCs) using confocal laser scanning microscopy (CLSM). First, we applied the 3D texture mapping method to render the volume of entire tissue sections. Then, we determined the chromatin texture quantitatively by calculating 3D gray-level co-occurrence matrices (3D GLCM) and 3D run length matrices (3D GLRLM). Finally, to demonstrate the suitability of 3D texture features for grading, we performed a discriminant analysis. In addition, we conducted a principal component analysis to obtain optimized texture features. Automatic grading of cell nuclei using 3D texture features had an accuracy of 78.30%. Combining 3D textural and 3D morphological features improved the accuracy to 82.19%. As a comparative study, we also performed a stepwise feature selection. Using the 4 optimized features, we could obtain more improved accuracy of 84.32%. Three dimensional texture features have potential for use as fundamental elements in developing a new nuclear grading system with accurate diagnosis and predicting prognosis.

• Environmental Engineering Research
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• 제24권3호
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• pp.382-388
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• 2019

Although nanobubbles attract significant attention, their characteristics and applications have not been thoroughly defined. There are diverse opinions about the definition of nanobubbles and controversy regarding methods that verify their characteristics. This study defines nanobubbles as having a size less than $1{\mu}m$. The generation of these sub-micron (nano) bubbles may be verified by induced coalescence or light scattering. The size of a sub-micron (nano) bubbles may be measured by optical, and confocal laser scanning microscopy. Also, the size may be estimated by the relationship of bubble size with the dissolved oxygen concentration. However, further research is required to accurately define the average bubble size. The zeta potential of sub-micron (nano) bubbles decreases as pH increases, and this trend is consistent for micron bubbles. When the bubble size is reduced to about 700-900 nm, they become stationary in water and lose buoyancy. This characteristic means that measuring the concentration of sub-micron (nano) bubbles by volume may be possible by irradiating them with ultrasonic waves, causing them to merge into micron bubbles. As mass transfer is a function of surface area and rising velocity, this strongly indicates that the application of sub-micron (nano) bubbles may significantly increase mass transfer rates in advanced oxidation and aeration processes.

• 대한소아치과학회지
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• 제51권1호
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• pp.32-39
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• 2024

This study evaluated the additive impact of ethylenediamine tetraacetic acid (EDTA) on erythrosine-mediated photodynamic therapy (PDT) against Streptococcus mutans (S. mutans) biofilm by measuring colony-forming units and applying confocal laser scanning microscopy. Fifty-six bovine incisors, free from dental caries or structural defects, were utilized in this study. Dentin specimens were created by cutting with a low-speed diamond disk under a continuous flow of water, resulting in dimensions of 6.0 mm × 3.0 mm × 2.0 mm. The specimens were categorized into 4 groups: Control, EDTA, PDT, and EDTA + PDT. S. mutans ATCC 25175 was employed to establish biofilm on the dentin specimens. A 17% EDTA solution was applied for 1 min. For PDT, erythrosine served as the photosensitizer. Finally, a light-emitting diode source (385 - 515 nm) was employed in this study. The PDT group exhibited a significantly lower bacterial count than both the control and EDTA groups (p < 0.001). The EDTA + PDT group demonstrated a significantly reduced bacterial count compared to the other 3 groups (p < 0.001). This study demonstrated that EDTA enhances the antimicrobial efficacy of PDT on S. mutans biofilm. Even at a low concentration of photosensitizer, the combination of EDTA and PDT yields a significant antibacterial effect.

• JSTS:Journal of Semiconductor Technology and Science
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• 제1권4호
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• pp.239-247
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• 2001

The Fluorescence-Activated Cell Sorter (FACS) is a well-established instrument used for identifying, enumerating, classifying and sorting cells by their physical and optical characteristics. For a miniaturized FACS device, a disposable plastic microchip has been developed which has a hydrodynamic focusing chamber using soft lithography. As the characteristics of the spatially confined sample stream have an effect on sample throughput, detection efficiency, and the accuracy of cell sorting, systematic fluid dynamic studies are required. Flow visualization is conducted with a laser scanning confocal microscopy (LSCM), and three-dimensional flow structure of the focused sample stream is reconstructed from 2D slices acquired at $1\mutextrm{m}$ intervals in depth. It was observed that the flow structure in the focusing chamber is skewed by unsymmetrical velocity profile arising from trapezoidal cross section of the microchannel. For a quantitative analysis of a microscopic flow structure, Confocal Micro-PIV system has been developed to evaluate the accelerated flow field in the focusing chamber. This study proposes a method which defines the depth of the measurement volume using a detection pinhole. The trajectories of red blood cells (RBCs) and their interactions with surrounding flow field in the squeezed sample stream are evaluated to find optimal shape of the focusing chamber and fluid manipulation scheme for stable cell transporting, efficient detection, and sorting

• Restorative Dentistry and Endodontics
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• 제43권1호
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• pp.2.1-2.10
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• 2018

Objectives: To determine the effect of size and insertion depth of irrigation needle on the amount of apical extruded debris and the amount of penetration depth of sealer using a confocal laser scanning microscope (CLSM). Materials and Methods: Twenty maxillary premolars were assigned to 2 groups (n = 10), according to the size of needle tip, 28 G or 30 G. Buccal roots of samples were irrigated with respective needle type inserted 1 mm short of the working length (WL), while palatal roots were irrigated with respective needle type inserted 3 mm short of the WL. Prepared teeth were removed from the pre-weighed Eppendorf tubes. Canals were filled with F3 gutta-percha cone and rhodamine B dye-labeled AH 26 sealer. Teeth were transversally sectioned at 1 and 3 mm levels from the apex and observed under a CLSM. Eppendorf tubes were incubated to evaporate the irrigant and were weighed again. The difference between pre- and post-weights was calculated, and statistical evaluation was performed. Results: Inserting needles closer to the apex and using needles with wider diameters were associated with significantly more debris extrusion (p < 0.05). The position of needles and level of sections had statistically significant effects on sealer penetration depth (p < 0.05 for both). Conclusions: Following preparation, inserting narrower needles compatible with the final apical diameter of the prepared root canal at 3 mm short of WL during final irrigation might prevent debris extrusion and improve sealer penetration in the apical third.

• 생명과학회지
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• 제22권10호
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• pp.1415-1419
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• 2012

Thymosin ${\beta}4$ 는 대장암에서 암 줄기세포 마커인 CD133을 지닌 세포에서 지니지 않은 세포에 비해 강하게 발현된다고 보고되어 있다. 본 연구에서는 thymosin ${\beta}4$와 줄기세포 마커인 CD133의 상관관계를 정상 위 조직에서 관찰하였다. Thymosin ${\beta}4$와 CD133의 발현 양상은 tissue microarray 조직상에서 면역화학적 방법으로 관찰하였으며 thymosin ${\beta}4$와 CD133의 존재 위치는 면역형광 염색법 및 confocal microscope를 사용하여 조사하였다. Thymosin ${\beta}4$와 CD133은 동일한 양상으로 발현되었으며 모두 위의 선조직에서 강하게 발현되었다. 면역 형광 염색법으로 두가지 단백질을 동시에 염색한 결과 두 단백질이 동일한 위치에서 함께 존재하는 것으로 규명되었다. 이러한 결과는 thymosin ${\beta}4$와 CD133은 정상위의 선조직에서 발현되며 두 단백질의 발현 양상 및 위치가 동일하여 서로 긴밀한 상호작용을 할 가능성을 제시하고 있다.