• Title/Summary/Keyword: fluorescence imaging

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Applicability of Fluorescein Diacetate (FDA) and Calcein-AM to Determine the Viability of Marine Plankton (FDA와 Calcein-AM 방법을 이용한 해양플랑크톤 생사판별기법)

  • Baek, Seung-Ho;Shin, Kyoung-Soon
    • Ocean and Polar Research
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    • v.31 no.4
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    • pp.349-357
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    • 2009
  • Ballast water is widely recognized as a serious environmental problem due to the risk of introducing non-indigenous aquatic species. In this study we aimed to investigate measures which can minimize the transfer of aquatic organisms from ballast water. Securing more reliable technologies to determine the viability of aquatic organisms is an important initiative in ballast water management systems. To evaluate the viability of marine phytoplankton, we designed the staining methods of fluorescein diacetate (FDA) and Calcein-AM assay on each target species belonging to different groups, such as bacillariphyceae, dinophyceae, raphidophyceae, chrysophyceae, haptophyceae and chlorophyceae. The FDA method, which is based on measurements of cell esterase activity using a fluorimetric stain, was the best dye for determining live cells of almost all phytoplankton species, except several diatoms tested in this study. On the other hand, although fluorescence of Calcein-AM was very clear for a comparatively longer time, green fluorescence per cell volume was lacking in most of the tested species. According to the Flow CAM method, which is a continuous imaging technique designed to characterize particles, green fluorescence values of stained cells by FDA were significantly higher than those of Calcein-AM treatments and control, implying that the Flow CAM using FDA assay could be adapted as an important tool for distinguishing living cells from dead cells. Our results suggest that the FDA and Calcein-AM methods can be adapted for use on phytoplankton, though species-specific characters are greatly different from one organism to another.

Planar Imaging of Temperature and Concentration of a Laminar Nonpremixed $H_2$/$N_2$flame Using a Tunable KrF Excimer Laser (파장 가변형 KrF 에시머 레이저를 이용한 층류 비예혼합 수소 화염에서의 2차원적 온도 및 농도 계측)

  • Kim, Gun-Hong;Jin, Seong-Ho;Kim, Yong-Mo;Park, Gyeong-Seok;Kim, Se-Won;Kim, Gyeong-Su
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.24 no.12
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    • pp.1580-1587
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    • 2000
  • Rayleigh scattering and laser induced predissociative fluorescence are employed for capturing two-dimensional images of temperature and species concentration in a laminar nonpremixed flame of a diluted hydrogen jet. Rayleigh scattering cross-sections are experimentally obtained ar 248nm. Dispersed LIPF spectra of OH and O$_2$ are also measured in a flame in order to confirm the excitation of single vibronic state of OH and O$_2$ .OH and O$_2$ are excited on the P$_1$(8) line of the A $^2\Sigma ^+(v^`=3) - X^2\pi (V^"=0)$ band and R(17) line of the Schumann-Runge band B $^3\Sigma _u^-(v^`=0) - X ^3\Sigma _g^-(v^"=6)$, respectively. Fluorescence spectra of OH and Hot O$_2$ are captured and two-dimensional images of the hydrogen flame field are successfully visualized.

Water-Soluble Distyrylbenzene Fluorophore and Fluorescence Behavior in a Polymeric Vesicle

  • Nayak, Rati Ranjan;Woo, Han-Young
    • Journal of the Korean Chemical Society
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    • v.51 no.6
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    • pp.513-519
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    • 2007
  • A vesicle forming polymer, poly(sodium acrylamidoundecanoate) (PSAU) and a water-soluble distyrylbenzene- based fluorophore, TPADSB-C were synthesized and characterized by using UV-vis and photoluminescence (PL) spectroscopy. An inter-chain vesicle formation of PSAU was observed at ~0.01 g/L from N-phenyl naphthylamine fluorescence measurement with changing PSAU concentration in water. Above critical aggregation concentration of PSAU, optical properties of TPADSB-C were investigated to study the microenvironment modulation through dye incorporation in the polymeric vesicle. The emission of TPADSB-C in the presence of PSAU vesicles was blue-shifted and the PL quantum efficiency was increased to 90% due to the microenvironment (e.g. polarity) change in aqueous solution. This study shows that the polymeric vesicle containing molecular fluorophores has a great potential as an efficient, stable and biocompatible labeling tag in biological cell imaging.

Development of Dental Calculus Diagnosis System using Fluorescence Detection (형광 검출을 이용한 치석 진단 시스템 개발)

  • Jang, Seon-Hui;Lee, Young-Rim;Lee, Woo-Cheol
    • The Journal of the Korea institute of electronic communication sciences
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    • v.17 no.4
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    • pp.715-722
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    • 2022
  • If you don't regularly go to the dentist to check your teeth, it is difficult to notice cavities or various diseases of your teeth until you have pain or discomfort. Dental plaque is produced by the combination of food or foreign substances and bacteria in the mouth. Starch breaks down from the bacteria that form tartar. The acid that occurs at this time melts the enamel of the teeth and becomes a cavity. So tartar management is important. Poppyrin, the metabolism of bacteria in the mouth, reacts at 405 nm wavelengths and becomes red fluorescent, which can be seen by imaging through certain wavelength filters. By the above method, Frag and tartar are fluorescently detected and photographed with a yellow series of filters that pass wavelengths of 500 nm or more. It uses MATLAB to detect and display red fluorescence through image processing. Using the difference in voltage between normal teeth and tartar through an optical measuring circuit, it was connected to an Arduino and displayed on the LCD. This allows the user to know the presence and location of dental plaque more accurately.

Design of Fluorescence Multi-cancer Diagnostic Sensor Platform based on Microfluidics (미세 유체 기반의 형광 다중 암 진단 센서 플랫폼 설계)

  • Lee, B.K.;Khaliq, A.;Jeong, M.Y.
    • Journal of the Microelectronics and Packaging Society
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    • v.29 no.4
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    • pp.55-61
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    • 2022
  • There is a major interest in diagnostic technology for multiple cancers worldwide. In order to reduce the difficulty of cancer diagnosis, a liquid biopsy technology based on a microfluidic device using trace amounts of biofluids such as blood is being studied. And optical biosensing, which measures the concentration of analytes through fluorescence imaging using biofluids, requires various strategies to improve sensitivity, and specialists and equipment are needed to carry out these strategies. This leads to an increase in diagnostic and production costs, and it is necessary to develop a technology to solve this problem. In this paper, we design and propose a fluorescent multi-cancer diagnostic sensing platform structure that implements passive self-separation technology and molecular recognition activation functions by fluid mixing, only with the geometry and microfluidic phenomena of microchannels based on self-driven flow by capillary force. In order to check the parameters affecting the performance of the plasma separation part of the designed sensor, the hydrodynamic diameter of the channel and the viscosity of the fluid were set as variables to confirm the formation of plasma separation flow through simulation. And finally, we propose an optimal sensor platform structure.

Application of Near-Infrared Fluorescence Imaging with Indocyanine Green in Totally Laparoscopic Distal Gastrectomy

  • Liu, Maoxing;Xing, Jiadi;Xu, Kai;Yuan, Peng;Cui, Ming;Zhang, Chenghai;Yang, Hong;Yao, Zhendan;Zhang, Nan;Tan, Fei;Su, Xiangqian
    • Journal of Gastric Cancer
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    • v.20 no.3
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    • pp.290-299
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    • 2020
  • Purpose: Recently, totally laparoscopic gastrectomy has been gradually accepted by surgeons worldwide for gastric cancer treatment. Complete dissection of the lymph nodes and the establishment of the surgical margin are the most important considerations for curative gastric cancer surgery. Previous studies have demonstrated that indocyanine green (ICG)-traced laparoscopic gastrectomy significantly improves the completeness of lymph node dissection. However, it remains difficult to identify the tumor location intraoperatively for gastric cancers that are staged ≤T3. Here, we investigated the feasibility of ICG fluorescence for lymph node mapping and tumor localization during totally laparoscopic distal gastrectomy. Materials and Methods: Preoperative and perioperative data from consecutive patients with gastric cancer who underwent a totally laparoscopic distal gastrectomy were collected and analyzed. The patients were categorized into the ICG (n=61) or the non-ICG (n=75) group based on whether preoperative endoscopic mucosal ICG injection was performed. Results: The ICG group had a shorter operation time and less intraoperative blood loss. Moreover, significantly more lymph nodes were harvested in the ICG group than the non-ICG group. No pathologically positive margin was found and there was no significant difference in either the proximal or distal surgical margins between the 2 groups. Conclusions: Near-infrared fluorescence imaging with ICG can be successfully used in totally laparoscopic distal gastrectomy, and it contributes to both the completeness of D2 lymph node dissection and confirmation of the gastric transection line. Well-designed prospective randomized studies are needed in the future to fully validate our findings.

Nondestructive sensing technologies for food safety

  • Kim, M.S.;Chao, K.;Chan, D.E.;Jun, W.;Lee, K.;Kang, S.;Yang, C.C.;Lefcourt, A.M.
    • 한국환경농학회:학술대회논문집
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    • 2009.07a
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    • pp.119-126
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    • 2009
  • In recent years, research at the Environmental Microbial and Food Safety Laboratory (EMFSL), Agricultural Research Service (ARS) has focused on the development of novel image-based sensing technologies to address agro-food safety concerns, and transformation of these novel technologies into practical instrumentation for industrial implementations. The line-scan-based hyperspectral imaging techniques have often served as a research tool to develop rapid multispectral methods based on only a few spectral bands for rapid online applications. We developed a newer line-scan hyperspectral imaging platform for high-speed inspection on high-throughput processing lines, capable of simultaneous multiple inspection algorithms for different agro-food safety problems such as poultry carcass inspection for wholesomeness and apple inspection for fecal contamination and defect detection. In addition, portable imaging devices were developed for in situ identification of contamination sites and for use by agrofood producer and processor operations for cleaning and sanitation inspection of food processing surfaces. The aim of this presentation is to illustrate recent advances in the above agro.food safety sensing technologies.

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Detection of E.coli biofilms with hyperspectral imaging and machine learning techniques

  • Lee, Ahyeong;Seo, Youngwook;Lim, Jongguk;Park, Saetbyeol;Yoo, Jinyoung;Kim, Balgeum;Kim, Giyoung
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.645-655
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    • 2020
  • Bacteria are a very common cause of food poisoning. Moreover, bacteria form biofilms to protect themselves from harsh environments. Conventional detection methods for foodborne bacterial pathogens including the plate count method, enzyme-linked immunosorbent assays (ELISA), and polymerase chain reaction (PCR) assays require a lot of time and effort. Hyperspectral imaging has been used for food safety because of its non-destructive and real-time detection capability. This study assessed the feasibility of using hyperspectral imaging and machine learning techniques to detect biofilms formed by Escherichia coli. E. coli was cultured on a high-density polyethylene (HDPE) coupon, which is a main material of food processing facilities. Hyperspectral fluorescence images were acquired from 420 to 730 nm and analyzed by a single wavelength method and machine learning techniques to determine whether an E. coli culture was present. The prediction accuracy of a biofilm by the single wavelength method was 84.69%. The prediction accuracy by the machine learning techniques were 87.49, 91.16, 86.61, and 86.80% for decision tree (DT), k-nearest neighbor (k-NN), linear discriminant analysis (LDA), and partial least squares-discriminant analysis (PLS-DA), respectively. This result shows the possibility of using machine learning techniques, especially the k-NN model, to effectively detect bacterial pathogens and confirm food poisoning through hyperspectral images.

Poloxamer 407 Hydrogels for Intravesical Instillation to Mouse Bladder: Gel-Forming Capacity and Retention Performance

  • Kim, Sang Hyun;Kim, Sung Rae;Yoon, Ho Yub;Chang, In Ho;Whang, Young Mi;Cho, Min Ji;Kim, Myeong Joo;Kim, Soo Yeon;Lee, Sang Jin;Choi, Young Wook
    • The Korean Journal of Urological Oncology
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    • 제15권3호
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    • pp.178-186
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    • 2017
  • Purpose: Poloxamer 407 (P407) thermo-sensitive hydrogel formulations were developed to enhance the retention time in the urinary bladder after intravesical instillation. Materials and Methods: P407 hydrogels (P407Gels) containing 0.2 w/w% fluorescein isothiocyanate dextran (FD, MW 4 kDa) as a fluorescent probe were prepared by the cold method with different concentrations of the polymer (20, 25, and 30 w/w%). The gel-forming capacities were characterized in terms of gelation temperature (G-Temp), gelation time (G-Time), and gel duration (G-Dur). Homogenous dispersion of the probe throughout the hydrogel was observed by using fluorescence microscopy. The in vitro bladder simulation model was established to evaluate the retention and drug release properties. P407Gels in the solution state were administered to nude mice via urinary instillation, and the in vivo retention behavior of P407Gels was visualized by using an in vivo imaging system (IVIS). Results: P407Gels showed a thermo-reversible phase transition at $4^{\circ}C$ (refrigerated; sol) and $37^{\circ}C$ (body temperature; gel). The G-Temp, G-Time, and G-Dur of FD-free P407Gels were approximately $10^{\circ}C-20^{\circ}C$, 12-30 seconds, and 12-35 hours, respectively, and were not altered by the addition of FD. Fluorescence imaging showed that FD was spread homogenously in the gelled P407 solution. In a bladder simulation model, even after repeated periodic filling-emptying cycles, the hydrogel formulation displayed excellent retention with continuous release of the probe over 8 hours. The FD release from P407Gels and the erosion of the gel, both of which followed zero-order kinetics, had a linear relationship ($r^2=0.988$). IVIS demonstrated that the intravesical retention time of P407Gels was over 4 hours, which was longer than that of the FD solution (<1 hour), even though periodic urination occurred in the mice. Conclusions: FD release from P407Gels was erosion-controlled. P407Gels represent a promising system to enhance intravesical retention with extended drug delivery.

PET Imaging of Click-engineered PSMA-targeting Immune Cells in Normal Mice

  • Hye Won Kim;Won Chang Lee;In Ho Song;Hyun Soo Park;Sang Eun Kim
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.8 no.2
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    • pp.53-61
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    • 2022
  • This study aimed to increase the targeting ability against PSMA in cell therapy using metabolic glycoengineering and biorthogonal chemistry and to visualize cell trafficking using PET imaging. Cellular membranes of THP-1 cells were decorated with azide(-N3) using Ac4ManNAz by metabolic glycoengineering. Engineered THP-1 cells were conjugated with DBCO-bearing fluorophore (ADIBO-Cy5.5) for 1 h at different concentrations and analyzed by confocal fluorescence microscopy and flow cytometry. For PSAM ligand conjugation to THP-1 cells, Ac4ManNAz treated THP-1 cells were incubated with DBCO-PSMA ligand (ADIBO-GUL) at a final concentration with 100 µM for 1 h. To evaluate the effect on cell recognition, PSMA ligand conjugated THP-1 cells(as effectors) were co-cultured with PSMA positive 22RV1 (as target cells) at 3 : 1 a effector-to-target cell (E/T) ratio. The interaction between THP-1 and 22RV1 was monitored by confocal fluorescence microscopy. For preparing the radiolabeled THP-1, the cells were treated at the activity of ~ 740 kBq of [89Zr]Zr(oxinate)4/5 × 106 cells. Radiolabeled cells were analyzed for determination of cell-associated radioactivity by gamma counting and viability using MTS assay. In the cytotoxicity assay, THP-1 cells did not have any cytotoxicity even when the Ac4ManNAz concentration was 100 µM. In confocal microscopy and flow cytometry, THP-1 cells were efficiently labeled ADIBO-Cy5.5 in a dose-dependent manner, and the dose of 100 µM was the optimal concentration for the following experiments. The clusters of PSMA ligand-conjugated THP-1 cells and 22RV1 cells were identified, indicating cell-cell recognition over the cell surface between two types of cells. Cell radiolabeling efficiency was 54.5 ± 17.8%. THP-1 labeled with 0.09 ± 0.03 Bq/cell showed no significant cytotoxicity compared to unlabeled THP-1 up to 7 days. We successfully demonstrated that Ac4ManNAz treated cells were efficiently conjugated with ADIBO-GUL for preparing the PSMA-targeting cells, and [89Zr]Zr(oxinate)4 could be used to label cells without toxicity. It suggested that PSMA-ligand conjugated cell therapy could be improved cell targeting and be monitored by PET imaging.