• Title/Summary/Keyword: Fluorescence detection system

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Discriminant analysis of grain flours for rice paper using fluorescence hyperspectral imaging system and chemometric methods

  • Seo, Youngwook;Lee, Ahyeong;Kim, Bal-Geum;Lim, Jongguk
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.633-644
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    • 2020
  • Rice paper is an element of Vietnamese cuisine that can be used to wrap vegetables and meat. Rice and starch are the main ingredients of rice paper and their mixing ratio is important for quality control. In a commercial factory, assessment of food safety and quantitative supply is a challenging issue. A rapid and non-destructive monitoring system is therefore necessary in commercial production systems to ensure the food safety of rice and starch flour for the rice paper wrap. In this study, fluorescence hyperspectral imaging technology was applied to classify grain flours. Using the 3D hyper cube of fluorescence hyperspectral imaging (fHSI, 420 - 730 nm), spectral and spatial data and chemometric methods were applied to detect and classify flours. Eight flours (rice: 4, starch: 4) were prepared and hyperspectral images were acquired in a 5 (L) × 5 (W) × 1.5 (H) cm container. Linear discriminant analysis (LDA), partial least square discriminant analysis (PLSDA), support vector machine (SVM), classification and regression tree (CART), and random forest (RF) with a few preprocessing methods (multivariate scatter correction [MSC], 1st and 2nd derivative and moving average) were applied to classify grain flours and the accuracy was compared using a confusion matrix (accuracy and kappa coefficient). LDA with moving average showed the highest accuracy at A = 0.9362 (K = 0.9270). 1D convolutional neural network (CNN) demonstrated a classification result of A = 0.94 and showed improved classification results between mimyeon flour (MF)1 and MF2 of 0.72 and 0.87, respectively. In this study, the potential of non-destructive detection and classification of grain flours using fHSI technology and machine learning methods was demonstrated.

Production of chickens with green fluorescent protein-knockin in the Z chromosome and detection of green fluorescent protein-positive chicks in the embryonic stage

  • Kyung Soo Kang;Seung Pyo Shin;In Su Ha;Si Eun Kim;Ki Hyun Kim;Hyeong Ju Ryu;Tae Sub Park
    • Animal Bioscience
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    • v.36 no.6
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    • pp.973-979
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    • 2023
  • Objective: The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system, which is the most efficient and reliable tool for precisely targeted modification of the genome of living cells, has generated considerable excitement for industrial applications as well as scientific research. In this study, we developed a gene-editing and detection system for chick embryo sexing during the embryonic stage. Methods: By combining the CRISPR/Cas9 technical platform and germ cell-mediated germline transmission, we not only generated Z chromosome-targeted knockin chickens but also developed a detection system for fluorescence-positive male chicks in the embryonic stage. Results: We targeted a green fluorescent protein (GFP) transgene into a specific locus on the Z chromosome of chicken primordial germ cells (PGCs), resulting in the production of ZGFP-knockin chickens. By mating ZGFP-knockin females (ZGFP/W) with wild males (Z/Z) and using a GFP detection system, we could identify chick sex, as the GFP transgene was expressed on the Z chromosome only in male offspring (ZGFP/Z) even before hatching. Conclusion: Our results demonstrate that the CRISPR/Cas9 technical platform with chicken PGCs facilitates the production of specific genome-edited chickens for basic research as well as practical applications.

DEVELOPMENT OF FILTERING SYSTEM OF LASER FLUORESCENCE FOR IMPROVEMENT OF THE DIAGNOSTIC SENSITIVITY FOR DENTAL CARIES (레이저 형광법의 치아우식증 진단감도를 증진시키기 위한 필터링 시스템 개발)

  • Lee, Sang-Ho;Park, Jong-Whi;Kook, Jung-Ki;Lee, Nan-Young
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.2
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    • pp.189-195
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    • 2003
  • The objective of this study is to improve the optical sensitivity of laser fluorescence for detection of incipient enamel caries. An incipient carious lesion was formed in various stages by placing an enamel specimen of a bovine tooth in STPP demineralization solution. After measuring the optical density of the lesion surface by laser fluorescence induced by argon laser and various alter of yellow(500-520nm), amber(520-540nm), orange(540-560nm), and red(560-580nm), the specimen was cut vertically to measure the depth of the lesion using a polarizing microscope. SAS statistical program was used to analyze the relationship between the optical density of the lesion suface and the depth of the lesion. The results were as follows: 1. The optical density of early carious lesion, measured by laser fluorescence with amber and orange filter, and lesion depth observed by polarizing microscope, were increased as demineralization time increased. 2. The correlation coefficient between optical density of the lesion surface and the histological depth of the lesion was the highest in orange filter(r=0.49), followed by amber(r=0.32), yellow(r=0.13) and red(0.01). 3. Regression analysis showed that the most linear relationship between the optical density and the lesion depth was existed in orange filter group. In regard above results, laser fluorescence could be considered to be reliable for optical diagnosis of dental caries.

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Frequency of Chromosome Aberrations Detected by Fluorescence in Situ Hybridization Using Triple Chromosome-Specific Probes in o Healthy Korean Population (3중 염색체 probe를 이용한 FISH(fluorescence in situ hybridization)기법으로 분석한 정상인의 염색체 이상빈도)

  • 정해원;김수영;신은희
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.109-115
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    • 1998
  • Fluorscence in situ hybridization with chromosome-specific probe has been shown to be a valid and rapid method for detection of chromosome rearrangements induced by chemical and physical agents. This method is useful for quantifying structural aberrations, expecially for stable ones, such as translocation and insertion, which are difficult to detect with conventional method in human lymphocyte. In order to use the FISH method as a biodosimeter for monitoring human population exposed to various chemical and physical agent, baseline level of chromosome rearragement was established. Blood from forty four healthy adults were collected and analysed with whole chromosome-specific probes by human chromosome 1,2 and 4. The frequencies of stable translocation were 2.45 per 100 cell equivalent and those of insertion, color juction, acentric and dicentric were 0.32, 3.28, 0.23 and 0.27 per 100 cell equivalent respectively. The frequencies of chromosome rearragements increased with age in both sexes except for dicenrics. From above result, stable aberrations accumulate with age and it may reflect integrated lifetime exposure of adverse environment.

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Forensic STR Analysis of Mixed Chimerism after Allogeneic Bone Marrow Transplantation

  • Eom, Yong-Bin
    • Biomedical Science Letters
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    • v.16 no.3
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    • pp.193-196
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    • 2010
  • Multiplex PCR-based short tandem repeat (STR) analysis is considered as a good tool for monitoring bone marrow engraftment after sex-mismatched allogeneic transplantation and provides a sensitive and accurate assessment of the contribution of both donor and/or recipient cells in post-transplantation specimens. Forensic STR analysis and quantitative real time PCR are used to determine the proportion of donor versus recipient each contained within the total DNA. The STR markers were co-amplified in a single reaction by using commercial $PowerPlex^{(R)}$ 16 system and $AmpFISTR^{(R)}$ $Identifiler^{(R)}$ / $Yfiler^{(R)}$ PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI $PRIS^{(R)}$ 3100 Genetic Analyzer with capillary electrophoresis. The $GeneMapper^{TM}$ ID software were used for size calling and analysis of STR profiles. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA / Y Human Male DNA Quantification Kit The intent of this study was to analyze the ratio of donor versus recipient cells in the post-transplant peripheral blood, spleen, lung and kidney specimens. Specimens were taken from the traffic accident male victim who had been engrafted from bone marrow female donor. Blood and spleen specimens displayed female donor DNA profile. Kidney specimen showed male recipient DNA profile. Interestingly, lung tissue showed mixed profiles. The findings of this study indicate that the forensic STR analysis using fluorescence labeling PCR combined with capillary electrophoresis is quick and reliable enough to assess the ratio of donor versus recipient cells and to monitor the mixed chimeric patterns.

Spectrofluorimetric Determination of n-Octanol Based on its Ternary Complex with $Eu^{3+}$ and TTA ($Eu^{3+}$, TTA, 그리고 n-Octanol의 삼성분착물에 의한 n-Octanol의 분광형광분석법에 관한 연구)

  • Cha, Ki-Won;Park, Kwang-Won
    • Analytical Science and Technology
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    • v.10 no.6
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    • pp.433-438
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    • 1997
  • The enhancing effect of n-octanol on the fluorescence intensity of the $Eu^{3+}$-thenoyltrifluoroacetone(TTA) system in the presence of Triton X-100 was studied using spectrofluorometric method. This complex exhibited very intense $Eu^{3+}$ ion fluorescence at 619nm, when optically excited at 345nm. Optimum conditions for the determination of n-octanol have also been investigated. The calibration graph was linear over the range $1{\times}10^{-5}M{\sim}1{\times}10^{-7}M$ and the detection limit for n-octanol is $1{\times}10^{-9}M$. The result obtained in the analysis of the synthetic sample agreed with the known value in the error range and the relative standard deviation was ca. 3.5%.

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Fluorometric Quantitative Analysis of Al(III) Ion Using 5-Methoxy-2-phenyliminomethylphenol

  • Kim, Sun-Deuk;Lee, Hye-Won
    • Bulletin of the Korean Chemical Society
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    • v.30 no.5
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    • pp.1026-1030
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    • 2009
  • A novel Schiff base ligand (N, O system) 5-methoxy-2-phenyliminomethylphenol ($5-CH_3O-PMP$) was synthesized. Using the synthesized ligand as a fluorescent reagent, a fluorometric method was developed for the quantitative analysis of Al(III) ion. The quantitative analysis of Al(III) ion was performed by making the complex compound between Al(III) ion and $5-CH_3O-PMP$ in ethanol-water solution (85/15, v/v, pH 6.2). The excitation wavelength (${\lambda}em$) of the complex compound was 397 nm while the emmision wavelength (${\lambda}em$) was 498 nm. The quantitative analysis of Al(III) ion was carried out by estimating the fluorescence intensity. The various calibration curves were used for the quantitative analysis in the range of 0.27$\sim$27 ng/mL Al(III) ion concentrations. The detection limit was 0.027 ng/mL. Using the fluorometric method developed in this study, satisfying results were obtained from various samples such as tap water, hot spring water, river water, sea water and waste water, which contained considerable amounts of interfering ions.

Application of Organized Media for Rapid Spectrofluorimetric Determination of Trace Amounts of Cr(VI) in the Presence of Cr(III)

  • Madrakian, Tayyebeh;Afkhami, Abbas;Mohammadnejad, Masoumeh
    • Bulletin of the Korean Chemical Society
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    • v.30 no.6
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    • pp.1252-1256
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    • 2009
  • A simple, selective and sensitive fluorescence quenching method was developed to the determination Cr(VI). The method is based on the oxidation of $I^-\;to\;{{I_3}^-}$ by Cr(VI) in sulfuric acid solution followed by immediate formation of ion association compound between I3 − and rhodamine 6G in Tween-80 micellar media at room temperature. The influence of several surfactants on rhodamine 6G fluorescence signal was studied; particular attention was paid in the aggregation behavior of rhodamine 6G–Tween-80 system. The experimental parameters (e.g., type of surfactant, reagent concentration) were studied and the optimal conditions were established. The linear calibration graph was obtained in the range 2.0 - 100.0 ng m$L^{-1}$ Cr(VI). The detection limit of the method was 0.37 ng m$L^{-1}$. The relative standard deviation (R.S.D.) is less than 5% (n = 5). The efficiency of the method for the determination of Cr(VI) in the presence of Cr(III) in the sample was investigated. The method was applied successfully to the determination of Cr(VI) and total Cr in water, and liver tissue samples.

The Use of a Tobacco mosaic virus-Based Expression Vector System in Chrysanthemum

  • Park, Minju;Baek, Eseul;Yoon, Ju-Yeon;Palukaitis, Peter
    • The Plant Pathology Journal
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    • v.33 no.4
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    • pp.429-433
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    • 2017
  • Chrysanthemums (Chrysanthemum morifolium) are susceptible to tobacco mosaic virus (TMV). TMV-based expression vectors have been used in high-throughput experiments for production of foreign protein in plants and also expressing green fluorescent protein (GFP) to allow visualization of TMV movement. Here, we used TMV expressing the GFP to examine the infection of chrysanthemum by a TMV-based expression vector. Viral replication, movement and GFP expression by TMV-GFP were verified in upper leaves of chrysanthemums up to 73 days post inoculation (dpi) by RT-PCR. Neither wild-type TMV nor TMV-GFP induced symptoms. GFP fluorescence was seen in the larger veins of the inoculated leaf, in the stem above the inoculation site and in petioles of upper leaves, although there was no consistent detection of GFP fluorescence in the lamina of upper leaves under UV. Thus, a TMV-based expression vector can infect chrysanthemum and can be used for the in vivo study of gene functions.

Simultaneous Fluorimetric Determination of On-line Preconcentrated HANs, DCAD and TCAD by Using RPLC with a Postcolumn Derivatization System

  • Jung, Sung-Woon;Choi, Yong-Wook
    • Bulletin of the Korean Chemical Society
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    • v.34 no.6
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    • pp.1783-1790
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    • 2013
  • A simultaneous analytical method has been developed for the fluorimetric determination of haloacetonitriles (HANs) [dichloroacetonitrile (DCAN), trichloroacetonitrile (TCAN), dibromoacetonitrile (DBAN), haloacetamides [dichloroacetamide (DCAD), and trichloroacetamde (TCAD)] in drinking water by using the combined on-line perconcentration/reversed phase liquid chromatography (RPLC)-postcolumn detection system. This on-line perconcentration system was achieved by employing a precolumn packed with a commercial solid phase extraction (SPE) sorbent for the enrichment and purification of the target analytes. The haloacetonitriles and haloacetamides were separated on CN analytical column in a 7.5% methanol-0.02 M phosphate buffered mobile phase at pH 3. The column effluents were reacted with postcolumn reagents of ophthaldialdehyde (OPA) and sulfite ion at pH 11.5, to produce a highly fluorescent isoindole fluorophore, which were measured with a fluorescence detector. Under the optimized conditions for RPLC and the postcolumn derivatization system all of the coefficient of determination of the standard calibration curves for the target analytes were over 0.99 and had a linear range from 5 to 100 ${\mu}g/L$. The detection limits showed 1.6 ${\mu}g/L$ for DCAD, 0.1 ${\mu}g/L$ for TCAD, 0.6 ${\mu}g/L$ for DCAN, 1.6 ${\mu}g/L$ for TCAN and 1 ${\mu}g/L$ for DBAN, and the recoveries were ranged from 64 to 99% except for DCAD with precisions less than 4.9% in distilled water, and from 72(${\pm}4%$) to 116%(${\pm}2%$) in tap water.