• Applied Microscopy
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• v.51
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• pp.4.1-4.12
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• 2021

Fluorescence in situ hybridization (FISH) is a technique to visualize specific DNA/RNA sequences within the cell nuclei and provide the presence, location and structural integrity of genes on chromosomes. A confocal Whole Slide Imaging (WSI) scanner technology has superior depth resolution compared to wide-field fluorescence imaging. Confocal WSI has the ability to perform serial optical sections with specimen imaging, which is critical for 3D tissue reconstruction for volumetric spatial analysis. The standard clinical manual scoring for FISH is labor-intensive, time-consuming and subjective. Application of multi-gene FISH analysis alongside 3D imaging, significantly increase the level of complexity required for an accurate 3D analysis. Therefore, the purpose of this study is to establish automated 3D FISH scoring for z-stack images from confocal WSI scanner. The algorithm and the application we developed, SHIMARIS PAFQ, successfully employs 3D calculations for clear individual cell nuclei segmentation, gene signals detection and distribution of break-apart probes signal patterns, including standard break-apart, and variant patterns due to truncation, and deletion, etc. The analysis was accurate and precise when compared with ground truth clinical manual counting and scoring reported in ten lymphoma and solid tumors cases. The algorithm and the application we developed, SHIMARIS PAFQ, is objective and more efficient than the conventional procedure. It enables the automated counting of more nuclei, precisely detecting additional abnormal signal variations in nuclei patterns and analyzes gigabyte multi-layer stacking imaging data of tissue samples from patients. Currently, we are developing a deep learning algorithm for automated tumor area detection to be integrated with SHIMARIS PAFQ.

• The Journal of the Korean dental association
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• v.56 no.1
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• pp.8-16
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• 2018

Purpose: The aim of in vitro study was to compare the diagnostic accuracy to detect non-cavitated enamel caries on smooth surface by using four kinds of the QLF devices. Materials and Methods: A total of 52 human permanent premolars and molars were used. Fluorescence images were captured by the QLF devices (Inspektor Pro, QLF-D, Qraycam, and Qraypen). Fluorescence loss of the QLF was calculated. The severity of lesions was categorized into the following 3 scores using polarized light microscopy: normal (S), enamel demineralization to outer half of enamel (D1), and inner half of the enamel up to the dentin-enamel junction (D2). The Kruskal-Wallis test was used to compare the fluorescence loss among the QLF devices. Spearman rank correlation coefficient between histological scores and fluorescence loss of the devices was calculated. The sensitivity, specificity, and area under the receiver operating curve (AUROC) were calculated to compare their diagnostic accuracies. Results: The correlation coefficients between histological scores and the fluorescence loss of the devices showed 0.77 to 0.81 (P < 0.001). All histological scores, the fluorescence loss among the devices showed no statistical difference. Among the devices, sensitivity, specificity, and AUC values of the fluorescence loss showed 0.84 to 0.94, 0.76 to 0.90, and 0.90 to 0.92, respectively. Conclusions: All QLF devices had no difference with excellent diagnostic accuracies to detect non-cavitated enamel caries on smooth surface.

• Research in Plant Disease
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• v.7 no.2
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• pp.116-119
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• 2001

Witches' broom symptoms were firstly found on tubers of Solanum tuberosum cv, Deijima, showing dense growth of spindly sprouts in Cheju province, Korea. Plantlets from the diseased plants also produced the typical witches' broom symptoms, having densely-growing small leaves when they became adult plants. At the later stages the diseased leaves were blightened. Presence of phytoplasma in plant tissues was confirmed by DAPI-staining fluorescence microscopy and electron microscopy, exhibiting its localization in sieve tubes of stem, petiole, and midrib. This is the first report of potato witches' broom in Korea.

• Journal of Adhesion and Interface
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• v.1 no.1
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• pp.47-55
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• 2000

In this article, the characterization of the interface of hybrid composites was discussed. Interfacial interaction in organic/inorganic hybrid composites, especially silica-containing hybrids can be characterized by fluorescence spectroscopy, small angle X-ray scattering (SAXS), scanning electron microscopy (SEM), atomic force microscopy (AFM), and $^{29}Si$ NMR spectroscopy measurements.

• Journal of the korean academy of Pediatric Dentistry
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• v.34 no.2
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• pp.183-191
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• 2007

Through out the world dental caries seems to be decreased as it is difficult to make an accurate diagnosis for dental caries. The traditional diagnostic method which is probing and x-ray taking has many limitations to diagnose the early caries, so there were recommendations for the needs of new equipments such as laser fluorescence(LF), digital imaging fiber-optic trans-illumination(DIFOTI), and quantitative light fluorescence (QLF) which were developed from various study results. Also confocal laser scanning microscopy(CLSM) and ultrasonics are used for research progression. This study is to evaluate whether it is possible to monitor accurately for remineralization amount of enamel surface early caries using DIFOTI or LF After inducing artificial caries to bovine teeth to 10 participants remineralization was enhanced by 0 ppm and 500 ppm fluoride mouth rinse solution for 3 weeks. Then they were cross sectioned and analyzed using gold standard of the lesion depth measured by CLSM. The following results were obtained: 1. The measured percentage of light intensity(luminosity ratio) by DIFOTI increased with remineralization period, and showed significant reverse correlation with lesion depth measured by CLSM (p<0.01). 2. The measurement of laser fluorescence increased with remineralization period, and showed significant correlation with lesion depth measured by CLSM (p<0.01). 3. To the result for CLSM, 500 ppm fluoride mouth rinse group showed rapid rate for decreased tendency of lesion depth than 0 ppm fluoride mouth rinse group. In conclusion DIFOTI system was used to measure accurately for the remineralization amount of early surface caries, it is a very useful equipment to detect precisely the changes for early enamel caries remineralization during treatments.

• Applied Microscopy
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• v.48 no.4
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• pp.96-101
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• 2018

The release of nanoscale membrane-bound vesicles is common in all three domains of life. These vesicles are involved in a variety of biological processes such as cell-to-cell communication, horizontal gene transfer, and substrate transport. Prokaryotes including bacteria and archaea release membrane vesicles (MVs) (20 to 400 nm in diameter) into their extracellular milieu. In spite of structural differences in cell envelope, both Gram-positive and negative bacteria produce MVs that contain the cell membrane of each bacterial species. Archaeal MVs characteristically show surface-layer encircling the vesicles. Filamentous fungi and yeasts as eukaryotic microbes produce bilayered exosomes that have varying electron density. Microbes also form intracellular vesicles and minicells that are similar to MVs and exosomes in shape. Electron and fluorescence microscopy could reveal the presence of DNA in MVs and exosomes. Given the biogenesis of extracellular vesicles from the donor cell, in situ high-resolution microscopy can provide insights on the structural mechanisms underlying the formation and release of microbial extracellular vesicles.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2002.11a
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• pp.68.2-69
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• 2002

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2000.10a
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• pp.49.2-49
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• 2000

• BMB Reports
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• v.35 no.6
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• pp.562-567
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• 2002

In order to find the cellular interaction factors of the Heliothis armigera nuclear polyhedrosis virus capsid protein VP39, a Heliothis armigera cell cDNA library was constructed. Then VP39 was used as bait. The host actin gene was isolated from the cDNA library with the yeast two-hybrid system. This demonstrated that VP39 could interact with its host actin in yeast. In order to corroborate this interaction in vivo, the vp39 gene was fused with the green fluorescent protein gene in plasmid pEGFP39. The fusion protein was expressed in the Hz-AM1 cells under the control of the Autographa californica multiple nucleopolyhedrovirus immediate early gene promoter. The host actin was labeled specifically by the red fluorescence substance, tetramethy rhodamine isothicyanete-phalloidin. Observation under a fluorescence microscopy showed that VP39, which was indicated by green fluorescence, began to appear in the cells 6 h after being transfected with pEGFP39. Red actin cables were also formed in the cytoplasm at the same time. Actin was aggregated in the nucleus 9 h after the transfection. The green and red fluorescence always appeared in the same location of the cells, which demonstrated that VP39 could combine with the host actin. Such a combination would result in the actin skeleton rearrangement.

• BMB Reports
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• v.54 no.3
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• pp.157-163
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• 2021

The transient interactions between cellular components, particularly on membrane surfaces, are critical in the proper function of many biochemical reactions. For example, many signaling pathways involve dimerization, oligomerization, or other types of clustering of signaling proteins as a key step in the signaling cascade. However, it is often experimentally challenging to directly observe and characterize the molecular mechanisms such interactions-the greatest difficulty lies in the fact that living cells have an unknown number of background processes that may or may not participate in the molecular process of interest, and as a consequence, it is usually impossible to definitively correlate an observation to a well-defined cellular mechanism. One of the experimental methods that can quantitatively capture these interactions is through membrane reconstitution, whereby a lipid bilayer is fabricated to mimic the membrane environment, and the biological components of interest are systematically introduced, without unknown background processes. This configuration allows the extensive use of fluorescence techniques, particularly fluorescence fluctuation spectroscopy and single-molecule fluorescence microscopy. In this review, we describe how the equilibrium diffusion of two proteins, K-Ras4B and the PH domain of Bruton's tyrosine kinase (Btk), on fluid lipid membranes can be used to determine the kinetics of homodimerization reactions.