• Title/Summary/Keyword: Confocal fluorescent

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Assessment of Inactivation for Salmonella spp. on Chicken Meat using Confocal Laser Microscopy and Flow Cytometry (공초점 현미경 및 유세포 분류기를 이용한 계육에서의 Salmonella균 불활성화 평가)

  • Jang, Keum-Il;Chung, Duck-Hwa;Ha, Sang-Do;Kim, Keun-Sung;Lee, Kyu-Ho;Kim, Min-Gon;Kim, Cheorl-Ho;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.290-294
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    • 2006
  • Inactivation rates of Salmonella enteritidis in vitro and in vivo were assessed using confocal microscopy and flow cytometry. S. enteritidis was inactivated with 1% (w/v) trisodium phosphate (TSP) and live cells, and inactive cells were distinguished by staining with fluorescent probe, LIVE/DEAD BacLight Bacteria Viability stain. After TSP treatment for 1 min, most of Salmonella cells changed from green (live cells) fluorescence to red (inactive cells) fluorescence, indication of effective sanitizing. Inactivation efficiency and contamination sites of S. enteritidis on chicken skin by TSP treatment were assessed using confocal laser microscopy. Precise flow cytometry histograms for viability changes of S. enteritidis. after TSP treatments were obtained. Efficiency of various sanitizer treatments on foodborne pathogens could be assessed using this method.

Clinical Microscopy: Performance, Maintenance and Laser Safety (임상에서의 현미경: 작동, 유지보수 및 레이저 안전)

  • Lee, Tae Bok
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.2
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    • pp.125-133
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    • 2019
  • A microscope is the fundamental research and diagnostic apparatus for clinical investigation of signaling transduction, morphological changes and physiological tracking of cells and intact tissues from patients in the biomedical laboratory science. Proper use, care and maintenance of microscope with comprehensive understanding in mechanism are fully requested for reliable image data and accurate interpretation for diagnosis in the clinical laboratory. The standard operating procedure (SOP) for light microscopes includes performance procedure, brief information of all mechanical parts of microscopes with systematic troubleshooting mechanism depending on the laboratory capacity. Maintenance program encompasses cleaning objective, ocular lenses and inner optics; replacement and calibration of light source; XY sample stage management; point spread function (PSF) measurement for confocal laser scanning microscope (CLSM); quality control (QC) program in fluorescent microscopy; and systematic troubleshooting. Laser safety is one of the concern for medical technologists engaged in CLSM laboratory. Laser safety guideline based on the laser classification and risk level, and advisory lab wear for CLSM users are also expatiated in this overview. Since acquired image data presents a wide range of information at the moment of acquisition, well-maintained microscopes with proper microscopic maintenance program are impulsive for its interpretation and diagnosis in the clinical laboratory.

Comparative lectin binding patterns of Cochlodinium polykrikoides Margalef

  • Rhodes, Lesley L.;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • v.35 no.3
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    • pp.153-157
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    • 2000
  • Four different FITC-conjugated lectins were used to visually evaluate lectin binding activity by optical staining quality using confocal laser scanning microscopy (CLSM) of Cochzodinium polykrikoides in nature (wild type) and culture (cultured type). Cells from the field and cultures treated with ConA fluoresced only at the outer cell wall, and the abundance and distribution of the fluorescent signal were similar. Treatment with PWM and HPA did not elicit fluorescence at the cell surface, but the wild type exposed to HPA showed greater binding than did the cultured cells, possibly due to greater concentrations of glucosamine. The wild type cells treated with LBL lectin showed a strong green fluorescence on the cell surface, whereas cultured cells did not. Signal intensity and abundance were greater than for any other lectins tested in this study. These results suggest that wild type and cultured type are significantly different based on surface sugar production. In particular, the wild type cells apear richer in galactosamine-like moieties. Neither glucose nor mannose-like moieties were present in either wild types or cultured cells.

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Growth-Suppressing Activity of the Transfected Cx26 on BICR-M1Rk Breast Cancer Cell Line

  • Lee, Hae-Jung;Rhee, Seung-Keun
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.477-482
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    • 2011
  • There are accumulating evidences suggesting that connexin (Cx), a gap junction channel-forming protein, acts as a growth suppressor in various cancer cells, and this effect is attributeed to the gap junction-mediated intercellular communication (GJIC). In order to characterize the relationship between the growth-arresting activity of Cx26 and its cytoplasmic localizations after expression, we linked a nuclear export signal (NES) sequence to Cx26 cDNA before transfecting into a rat breast cancer cell line. A confocal fluorescent microscopic observation revealed that the insertion of NES minimized the nuclear expression of Cx26, and increased its cytoplasmic expression, including plasma membrane junctions. Total cell counting and BrdUrd-labeling experiments showed that the growth of the breast cancer cells was inhibited by 74% upon transfection of Cx26-NES, whereas only 9% inhibition was observed with only Cx26 cDNA.

In Vivo Evaluation of Multi Lamellar Vesicle Liposome’s Percutaneous Absorption and Stability

  • Joung, Min-Seok;Park, Jong-Oan;Seo, Bong-Seok;Ryu, Chang-Duck
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.27 no.1
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    • pp.29-42
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    • 2001
  • We had prepared MLV liposome with Hibiscus Esculentus Ext.(HEE) which have fluorescent light in order to evaluate its percutaneous absorption about hairless rat skin. Then we investigated particle size of MLV using confocal laser scanning microscope(CLSM) and transmission electron microscope(TEM), respectively. Stability of MLV liposome and penetration of MLV liposome to hairless rat skin was measured by CLSM. As a result of experiments, MLV was globular shape and the rage of particle size was 0.3-0.5$\mu\textrm{m}$ mostly. Cream-type MLV had high stability comparatively. When we treated with MLV to rat skin, skin penetration was enhanced, especially, the optimum concentration of MLV on penetration to rat skin was 10%. Optimum penetration time was 6hr-12hr. And MLV-type HEE was more effective on percutaneous absorption than HEE-cream or liposome-type HEE.

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THE ADAPTIBILITY OF LIGHT-CURED GLASS-IONOMER CEMENTS TO CAVITY WALLS. (와동벽에 대한 광중합형 글래스아이오노머 시멘트의 적합도)

  • Kang, Mi-Sun;Hur, Bock
    • Restorative Dentistry and Endodontics
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    • v.20 no.1
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    • pp.173-182
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    • 1995
  • The purpose of this study was to evaluate the adaptability of light-cured glass ionomer cement to cavity walls. Class V cavities were prepared on the labial surfaces of extracted bovine incisor teeth. The cavities were restored with Fuji II as self-cured glass ionomer cement and Fuji II LC, Vitremer as light-cured glass ionomer cement. Fluorescent markers (fluoreceine and rhodamin B) were incorperated into liquid and primer for a better image of microscopic observation. Restored teeth were sectioned by longitudinal and labiolingual direction. The adaptability at the tooth-restoration interface was assessed incisally, axially and cervically by confocal scanning laser microscope. Following results were obtained : 1. Chemical-cured glass iomomer cement restoration showed close adaptation on the all of the cavity walls, but, cracks formed within the cement. 2. Light-cured glass ionomer cement restoration was well adapted to the cavity walls, but showed crack in the cement adjacent to axial dentinal wall. 3. There' was no significant difference in adaptability between two light-cured glass ionomer cement restorations.

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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.

Applications of Optical Imaging System in Dentistry

  • Eom, Joo Beom;Park, Anjin
    • Medical Lasers
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    • v.9 no.1
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    • pp.25-33
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    • 2020
  • Optical-based imaging technology has high resolution and can assess images in real time. Numerous studies have been conducted for its application in the dental field. The current research introduces an oral camera that includes fluorescent imaging, a second study examining a 3D intraoral scanner applying a confocal method and a polarization structure that identifies the 3D image of a tooth, and finally, an optical coherence tomography technique. Using this technique, we introduce a new concept 3D oral scanner that simultaneously implements 3D structural imaging as well as images that diagnose the inside of teeth. With the development of light source technology and detector technology, various optical-based imaging technologies are expected to be applied in dentistry.

Fluorescent Method for Observing Intravascular Bonghan Duct (형광염색을 이용한 혈관내봉한관의 관찰)

  • Lee, Byung-Cheon;Baik, Ku-Youn;Johng, Hyeon-Min;Sung, Baeck-Kyoung;Soh, Kyung-Soon;Kang, Dae-In;Soh, Kwang-Sup
    • Journal of Pharmacopuncture
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    • v.8 no.3
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    • pp.5-9
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    • 2005
  • Observation of intra-vascular threadlike structures in the blood vessels of rats is reported with the images by differential interference contrast microscope, and fluorescence inverted microscope of the acridine-orange stained samples. The confocal microscope image and the hematoxylin-eosin staining revealed the distinctive pattern of nuclei distribution that clearly discerned the threadlike structure from fibrin, capillary, small venule, arteriole, or lymph vessel. Physiological function of the intra-vascular thread in connection with acupuncture is discussed. Especially, this threadlike duct can be a circulation path for herb-liquid flow, which may provide the scientific mechanism for therapeutic effect of herbal acupuncture.

Effect of Various Agents on Oral Bacterial Phagocytosis in THP-1 Cells

  • Song, Yuri;Lee, Hyun Ah;Na, Hee Sam;Jin, Chung
    • International Journal of Oral Biology
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    • v.43 no.4
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    • pp.217-222
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    • 2018
  • Phagocytosis is a fundamental process in which phagocytes capture and ingest foreign particles including pathogenic bacteria. Several oral pathogens have anti-phagocytic strategies, which allow them to escape from and survive in phagocytes. Impaired bacteria phagocytosis increases inflammation and contributes to inflammatory diseases. The purpose of this study is to investigate the influences of various agents on oral pathogenic phagocytosis. To determine phagocytosis, Streptococcus mutans, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis were stained with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE), and was measured using flowcytometery and confocal microscopy. The influencing factors on phagocytosis were evaluated through the pretreatment of ROS inhibitor (N-acetyl-L-cysteine (NAC)), lysozyme, potassium chloride (KCI) and adenosine triphosphate (ATP) in THP-1 cells. Expression of pro-inflammatory cytokines was determined by enzyme-linked immunosorbent assay (ELISA). The phagocytosis of various bacteria increased in a MOI-dependent manner. Among the tested bacteria, phagocytosis of P. gingivalis showed the highest fluorescent intensity at same infection time. Among the tested inhibitors, the NAC treatment significantly inhibited phagocytosis in all tested bacteria. In addition, NAC treatment indicated a similar pattern under the confocal microscopy. Moreover, NAC treatment significantly increased the bacteria-induced secretion of $IL-1{\beta}$ among the tested inhibitors. Taken together, we conclude that the phagocytosis occurs differently depending on each bacterium. Down-regulation by ROS production inhibited phagocytosis and lead increased of oral pathogens-associated inflammation.