• Title/Summary/Keyword: Fluorescence detection system

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A Linear Beacon System Featuring an Internal Deoxyguanine Quencher Allows Highly Selective Detection of Single Base Mismatches

  • Lee, Young-Ae;Hwang, Gil-Tae
    • Bulletin of the Korean Chemical Society
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    • v.31 no.7
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    • pp.2011-2014
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    • 2010
  • The fluorescence intensity of a single-stranded oligonucleotide containing a fluorene-labeled deoxyuridine $(U^{Fl})$ unit increases by only 1.5-fold upon formation of its perfectly matched duplex. To increase the fluorescence signal during hybridization, we positioned a quencher strand containing a deoxyguanine (dG) nucleobase, functioning as an internal quencher, opposite to the $U^{Fl}$ unit to reduce the intrinsic fluorescence upon hybridization with a probe. From an investigation of the optimal length of the quencher strand and the effect of the neighboring base sequence, we found that a short strand (five-nucleotide) containing all natural nucleotides and dG as an internal quencher was effective at reducing the intrinsic fluorescence of a linear beacon; it also exhibited high total discrimination factors for the formation of perfectly matched and single base-mismatched duplexes. Such assays that function based on clear changes in fluorescence in response to single-base nucleotide mutations would be useful tools for accelerating diagnoses related to various diseases.

Simultaneous Determination of Polycyclic Aromatic Hydrocarbons and Their Nitro-derivatives in Airborne Particulates by Using Two-dimensional High-performance Liquid Chromatography with On-line Reduction and Fluorescence Detection

  • Boongla, Yaowatat;Orakij, Walaiporn;Nagaoka, Yuuki;Tang, Ning;Hayakawa, Kazuichi;Toriba, Akira
    • Asian Journal of Atmospheric Environment
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    • v.11 no.4
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    • pp.283-299
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    • 2017
  • An analytical method using high-performance liquid chromatography (HPLC) with fluorescence (FL) detection was developed for simultaneously analyzing 10 polycyclic aromatic hydrocarbons (PAHs) and 18 nitro-derivatives of PAHs (NPAHs). The two-dimensional HPLC system consists of an on-line clean-up and reduction for NPAHs in the 1st dimension, and separation of the PAHs and the reduced NPAHs and their FL detection in the 2nd dimension after column-switching. To identify an ideal clean-up column for removing sample matrix that may interfere with detection of the analytes, the characteristics of 8 reversed-phase columns were evaluated. The nitrophenylethyl (NPE)-bonded silica column was selected because of its shorter elution band and larger retention factors of the analytes due to strong dipole-dipole interactions. The amino-substituted PAHs (reduced NPAHs), PAHs and deuterated internal standards were separated on polymeric octadecyl-bonded silica (ODS) columns and by dual-channel detection within 120 min including clean-up and reduction steps. The limits of detection were 0.1-9.2 pg per injection for PAHs and 0.1-140 pg per injection for NPAHs. For validation, the method was applied to analyze crude extracts of fine particulate matter ($PM_{2.5}$) samples and achieved good analytical precision and accuracy. Moreover, the standard reference material (SRM1649b, urban dust) was analyzed by this method and the observed concentrations of PAHs and NPAHs were similar to those in previous reports. Thus, the method developed here-in has the potential to become a standard HPLC-based method, especially for NPAHs.

Comparison of Clinical Characteristics of Fluorescence in Quantitative Light-Induced Fluorescence Images according to the Maturation Level of Dental Plaque

  • Jung, Eun-Ha;Oh, Hye-Young
    • Journal of dental hygiene science
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    • v.21 no.4
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    • pp.219-226
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    • 2021
  • Background: Proper detection and management of dental plaque are essential for individual oral health. We aimed to evaluate the maturation level of dental plaque using a two-tone disclosing agent and to compare it with the fluorescence of dental plaque on the quantitative light-induced fluorescence (QLF) image to obtain primary data for the development of a new dental plaque scoring system. Methods: Twenty-eight subjects who consented to participate after understanding the purpose of the study were screened. The images of the anterior teeth were obtained using the QLF device. Subsequently, dental plaque was stained with a two-tone disclosing solution and a photograph was obtained with a digital single-lens reflex (DSLR) camera. The staining scores were assigned as follows: 0 for no staining, 1 for pink staining, and 2 for blue staining. The marked points on the DSLR images were selected for RGB color analysis. The relationship between dental plaque maturation and the red/green (R/G) ratio was evaluated using Spearman's rank correlation. Additionally, different red fluorescence values according to dental plaque accumulation were assessed using one-way analysis of variance followed by Scheffe's post-hoc test to identify statistically significant differences between the groups. Results: A comparison of the intensity of red fluorescence according to the maturation of the two-tone stained dental plaque confirmed that R/G ratio was higher in the QLF images with dental plaque maturation (p<0.001). Correlation analysis between the stained dental plaque and the red fluorescence intensity in the QLF image confirmed an excellent positive correlation (p<0.001). Conclusion: A new plaque scoring system can be developed based on the results of the present study. In addition, these study results may also help in dental plaque management in the clinical setting.

A Skin Cancer Region Extraction Using Watershed (워터쉐드를 이용한 피부암 영역 추출)

  • Han, Jae-Bok;Kim, Jin-Young;Yu, Hong-Yeon;Hong, Sung-Hoon
    • Proceedings of the IEEK Conference
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    • 2006.06a
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    • pp.877-878
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    • 2006
  • In this paper, we propose a skin lesion detection to develop the system of fluorescence image analysis to identify the fluorescence of topical methyl aminolevulinate(MAL) idduced PpIX in patients with BCC accurately. By fluorescence image analysis we define the border between tumo and tumor-free areas on fluorescence image after topical application of MAL ointment. We excised both the tumor and peri-tumoral areas widely from the 10 patients with BCC, and divided tissue samples into 3 area, such as tumor area, suspected tumor area, tumor-free area, respectively. Our proposed method migt play a role as an adjunctive tool to define the border between tumor and tumor-free areas for Mohs' micrographic surgery.

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Review of Neurosurgical Fluorescence Imaging Systems for Clinical Application

  • Kim, Hong Rae;Lee, Hyun Min;Yoo, Heon;Lee, Seung Hoon;Kim, Kwang Gi
    • Journal of the Optical Society of Korea
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    • v.20 no.2
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    • pp.305-313
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    • 2016
  • A number of fluorescence imaging techniques for use in the surgical removal of glioma have been developed over the course of the long history of neurosurgery. Various biomarkers, biochemical agents, and detection systems for glioma have also been developed. This review focuses on 5-aminolevulinic acid (5-ALA), which is used to detect glioma. Numerous forms of fluorescence-guided surgery use 5-ALA, which is helpful to the surgeon. The surgical microscope system is the observational method generally used with 5-ALA, while the loupe, endoscope, and exoscope are simpler alternatives. A system is needed for minimal resection and other issues that arise during neurosurgery. Such an enhanced system should be able to detect low-grade tumors and provide information on microinvasive diseases, resulting in an improved survival rate and better surgical skills. Development of systems that fulfill certain needs would help protect the brain function of the patient and broaden the use of such systems in neurosurgery.

A New Reporter Vector System Based on Flow-Cytometry to Detect Promoter Activity

  • Jung, Sun-Do;Choi, Ji-Hye;Hong, Chang-Wan;Lee, Hyun-Ji;Park, Yoon-Kyung;Shin, Jung-Hoon;Park, Jae-Won;Park, Se-Ho
    • IMMUNE NETWORK
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    • v.9 no.6
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    • pp.243-247
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    • 2009
  • In this study, we report the development of a new dual reporter vector system for the analysis of promoter activity. This system employs green fluorescence emitting protein, EGFP, as a reporter, and uses red fluorescence emitting protein, DsRed, as a transfection control in a single vector. The expression of those two proteins can be readily detected via flow cytometry in a single analysis, with no need for any further manipulation after transfection. As this system allows for the simultaneous detection of both the control and reporter proteins in the same cells, only transfected cells which express the control protein, DsRed, can be subjected to promoter activity analysis, via the gating out of all un-transfected cells. This results in a dramatic increase in the promoter activity detection sensitivity. This novel reporter vector system should prove to be a simple and efficient method for the analysis of promoter activity.

Novel Cell-based Protease Assay System for Molecular Cell Biology and Drug Discovery

  • Hwang, Hyun-Jin;Kim, Jeong-Hee;Park, Joon-Woo;Kim, Sung-Hee;Lee, Min-Jeon;Jeong, Han-Seung;Hwang, In-Hwan
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.169.1-169.1
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    • 2003
  • Recently development of cell-based assay systems which are useful in molecular cell biology and drug discovery attracts significant attention. Here, we introduce a new technologies for monitoring enzyme activity and its inhibition inside living cells. Among various enzymes, proteases are important targets for studying various biological and disease-related processes such as viral infections, apoptosis and Alzheimer's disease. In this study, a sensitive cell-based protease detection system that enables direct fluorescence detection of a target protease and its inhibition inside living cells is introduced. (omitted)

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Development of Total Cholesterol Detection System by Fluorescence Chromatography (형광 크로마토그래피에 의한 콜레스테롤 측정법의 개발)

  • Oh, Sang-Wook
    • Journal of Food Hygiene and Safety
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    • v.24 no.2
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    • pp.148-153
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    • 2009
  • In this study, we have developed a fluorescence chromatographic assay for the quantification of total cholesterol in serum, which is a well-known risk predictor for cardiovascular diseases. The new assay system consists of a chromatographic strip in a cartridge, enzyme buffer containing cholesterol esterase, cholesterol oxidase, horseradish peroxidase, and color developer AEC, and a laser fluorescence scanner. The correlation coefficient (r) between cholesterol concentration and relative fluorescence units was 0.968 in the new assay, showing a reliable linearity through the tested range of cholesterol. Recovery test and comparability with a Hitachi 747 instrument showed 106.5-94% and r = 0.939 (p<0.001), respectively. The new assay system for cholesterol was developed as a pre-POCT platform conducted in clinics since it is fast (8 min) and uses a small volume of sample ($5\;{\mu}l$), and it may be applied for on-site diagnostics to replace expensive automated biochemical analyzer.

Development of Image-based System for Multiple Fluorescence Imaging Study (다중형광영상 연구를 위한 영상기반 시스템 개발)

  • Yoon, WoongBae;Kim, Hong Rae;Lee, Hyun Min;Kim, Young Jae;Kim, Kwang Gi;Yoo, Heon;Lee, Seung Hoon
    • Journal of Korea Multimedia Society
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    • v.18 no.12
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    • pp.1445-1452
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    • 2015
  • In these days, fluorescent materials such as ICG or 5-ALA is used for the brain surgery. The patients who underwent brain tumor surgery has been increased during last 30 years and the survivorship rate increased 22∼33% in 5 years. Recently, the Fluorescence induction surgery is developed for more safety and improved the resection rate for the glioma in the neurosurgery field. In this study, we proposed fluorescence area detection method for ICG and 5-ALA fluorescence induced surgery using acquired images from image processing. Accuracy was 99.21% from ICG images, and 99.51% from 5-ALA images. Matthews correlation coefficient was 88.67% from ICG images, and 90.49% from 5-ALA images.

Fluorescence photon counting rate as a function of dye concentration: Effect of dead time of photon detector (색소 농도에 따른 형광 광자의 계수율 : 광자 검출기의 dead time 효과)

  • 고동섭
    • Korean Journal of Optics and Photonics
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    • v.8 no.4
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    • pp.353-355
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    • 1997
  • A single molecule detection system, which consists of confocal fluorescence microscope and single photon counter, has been used to observe the dye concentration dependence of photon counting rate. With increasing concentration, a saturation effect of counting is observed and demonstrated on the basis of the dead time of photon detector. The equations presented here show the relations between the counting rate and some parameters such as probe volume, quantum efficiency of detector, and fluorescence photon number entered onto detector. The signal-to-noise ratio is also discussed briefly.

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