• Title/Summary/Keyword: real-time fluorescent imaging

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Real-Time Temporal Dynamics of Bicistronic Expression Mediated by Internal Ribosome Entry Site and 2A Cleaving Sequence

  • Lee, Soomin;Kim, Jeong-Ah;Kim, Hee-Dae;Chung, Sooyoung;Kim, Kyungjin;Choe, Han Kyoung
    • Molecules and Cells
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    • v.42 no.5
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    • pp.418-425
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    • 2019
  • Multicistronic elements, such as the internal ribosome entry site (IRES) and 2A-like cleavage sequence, serve crucial roles in the eukaryotic ectopic expression of exogenous genes. For utilization of multicistronic elements, the cleavage efficiency and order of elements in multicistronic vectors have been investigated; however, the dynamics of multicistronic element-mediated expression remains unclear. Here, we investigated the dynamics of encephalomyocarditis virus (EMCV) IRES- and porcine teschovirus-1 2A (p2A)-mediated expression. By utilizing real-time fluorescent imaging at a minute-level resolution, we monitored the expression of fluorescent reporters bridged by either EMCV IRES or p2A in two independent cultured cell lines, HEK293 and Neuro2a. We observed significant correlations for the two fluorescent reporters in both multicistronic elements, with a higher correlation coefficient for p2A in HEK293 but similar coefficients for IRES-mediated expression and p2A-mediated expression in Neuro2a. We further analyzed the causal relationship of multicistronic elements by convergent cross mapping (CCM). CCM revealed that in all four conditions examined, the expression of the preceding gene causally affected the dynamics of the subsequent gene. As with the cross correlation, the predictive skill of p2A was higher than that of IRES in HEK293, while the predictive skills of the two multicistronic elements were indistinguishable in Neuro2a. To summarize, we report a significant temporal correlation in both EMCV IRES- and p2A-mediated expression based on the simple bicistronic vector and real-time fluorescent monitoring. The current system also provides a valuable platform to examine the dynamic aspects of expression mediated by diverse multicistronic elements under various physiological conditions.

Multispectral intravital microscopy for simultaneous bright-field and fluorescence imaging of the microvasculature

  • Barry G. H. Janssen;Mohamadreza Najiminaini;Yan Min Zhang;Parsa Omidi;Jeffrey J. L. Carson
    • Applied Microscopy
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    • v.51
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    • pp.12.1-12.12
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    • 2021
  • Intravital video microscopy permits the observation of microcirculatory blood flow. This often requires fluorescent probes to visualize structures and dynamic processes that cannot be observed with conventional bright-field microscopy. Conventional light microscopes do not allow for simultaneous bright-field and fluorescent imaging. Moreover, in conventional microscopes, only one type of fluorescent label can be observed. This study introduces multispectral intravital video microscopy, which combines bright-field and fluorescence microscopy in a standard light microscope. The technique enables simultaneous real-time observation of fluorescently-labeled structures in relation to their direct physical surroundings. The advancement provides context for the orientation, movement, and function of labeled structures in the microcirculation.

A Rapid and Convenient Method for in Vivo Fluorescent Imaging of Protoscolices of Echinococcus multilocularis

  • Yang, Tao;Wang, Sibo;Zhang, Xuyong;Xia, Jie;Guo, Jun;Hou, Jixue;Zhang, Hongwei;Chen, Xueling;Wu, Xiangwei
    • Parasites, Hosts and Diseases
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    • v.54 no.2
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    • pp.225-231
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    • 2016
  • Human and animal alveolar echinococcosis (AE) are important helminth infections endemic in wide areas of the Northern hemisphere. Monitoring Echinococcus multilocularis viability and spread using real-time fluorescent imaging in vivo provides a fast method to evaluate the load of parasite. Here, we generated a kind of fluorescent protoscolices in vivo imaging model and utilized this model to assess the activity against E. multilocularis protoscolices of metformin (Met). Results indicated that JC-1 tagged E. multilocularis can be reliably and confidently used to monitor protoscolices in vitro and in vivo. The availability of this transient in vivo fluorescent imaging of E. multilocularis protoscolices constitutes an important step toward the long term bio-imaging research of the AE-infected mouse models. In addition, this will be of great interest for further research on infection strategies and development of drugs and vaccines against E. multilocularis and other cestodes.

A Study on PWM Control of Near-Infrared Fluorescence Imaging System (근적외선 형광 영상시스템의 PWM 제어에 관한 연구)

  • Lee, Byeong-Ho;Pan, Sung Bum
    • The Journal of Korean Institute of Information Technology
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    • v.16 no.11
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    • pp.115-121
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    • 2018
  • Fluorescent images using near-infrared light have no worry about radioactivity, and images can be checked in real time during surgery. Therefore experiments using fluorescent images for monitoring lymph node biopsy are actively under way. Fluorescent imaging equipment uses high heat-generating components such as LED and camera, thus uses water-cooling system as a stable heating suppression means. However in the fluorescent image equipment, the water cooling system takes a large volume which is a disadvantage in terms of miniaturization of the equipment. Even if the air cooling system is used for miniaturizing the equipment, heat generation is a problem. In this paper, we have experimented with the air cooling method using PWM control for the miniaturization of the equipment, and confirmed the constant quality of the fluorescent image and the suppression of the heat generation without any problems even when the equipment is used for a long time.

An image used by cross concept of two images - Gap of image space2 - (두가지 영상의 교차개념을 도입한 영상설치작품 - 상 공간의 틈새2 -)

  • Cho Ok-Hue;Lee Jun-Eui;Kim Hyung-Gi
    • Archives of design research
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    • v.19 no.4 s.66
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    • pp.205-208
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    • 2006
  • This work is a interactive digital art work which used the multiplex of jitter. It is an work which used a cross concept of two images. This work shows two kinds of images. One is on the wall through the gap of fluorescent lamp screen, and the other is on the fluorescent lamp screen. Two images are intercepted and comes out to the length as the multiplex of the jitter. The appearance of the spectator be recognized on the fluorescent lamp screen or on the wall, by two branch image which comes out intersecting of a length with multiplex function of the jitter. Two branch features of the image which it produces and the spectator which is input at real-time is equally divided, or in impression of juxtaposition goes in the spectator. It expresses the duplicity of the human being, with physical existence space divides the time space which we feel and there is a governing grammar which is different with each other and to express the thing it becomes. It used the image of the screen which it produces with the fluorescent lamp and cross concept.

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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.

Real-time Imaging of Inositol 1,4,5-trisphosphate Movement in Mouse Salivary Gland Cells

  • Hong, Jeong-Hee;Lee, Syng-Ill;Shin, Dong-Min
    • International Journal of Oral Biology
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    • v.33 no.4
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    • pp.125-129
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    • 2008
  • Inositol 1,4,5-trisphosphate ($IP_3$) plays an important role in the release of $Ca^{2+}$ from intracellular stores into the cytoplasm in a variety of cell types. $IP_3$ translocation dynamics have been studied in response to many types of cell signals. However, the dynamics of cytosolic $IP_3$ in salivary acinar cells are unclear. A green fluorescent protein (GFP)-tagged pleckstrin homology domain (PHD) was constructed and introduced into a phospholipase C ${\delta}1$ (PLC ${\delta}1$) transgenic mouse, and then the salivary acinar cells were isolated. GFP-PHD was heterogeneously localized at the plasma membrane and intracellular organelles in submandibular gland and parotid gland cells. Application of trypsin, a G protein-coupled receptor activator, to the two types of cells caused an increase in GFP fluorescence in the cell cytoplasm. The observed time course of trypsin-evoked $IP_3$ movement in acinar cells was independent of cell polarity, and the fluorescent label showed an immediate increase throughout the cells. These results suggest that GFP-PHD in many tissues of transgenic mice, including non-cultured primary cells, can be used as a model for examination of $IP_3$ intracellular dynamics.

Endoscopic Fluorescence Angiography with Indocyanine Green : A Preclinical Study in the Swine

  • Cho, Won-Sang;Kim, Jeong Eun;Kim, Sae Hoon;Kim, Hee Chan;Kang, Uk;Lee, Dae-Sic
    • Journal of Korean Neurosurgical Society
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    • v.58 no.6
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    • pp.513-517
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    • 2015
  • Objective : Microscopic indocyanine green (ICG) angiography is useful for identifying the completeness of aneurysm clipping and the preservation of parent arteries and small perforators. Neuroendoscopy is helpful for visualizing structures beyond the straight line of the microscopic view. We evaluated our prototype of endoscopic ICG fluorescence angiography in swine, which we developed in order to combine the merits of microscopic ICG angiography and endoscopy. Methods : Our endoscopic ICG system consists of a camera, a light source, a display and software. This system can simultaneously display real-time visible and near infrared fluorescence imaging on the same monitor. A commercially available endoscope was used, which was 4 mm in diameter and had an angle of $30^{\circ}$. A male crossbred swine was used. Results : Under general anesthesia, a small craniotomy was performed and the brain surface of the swine was exposed. ICG was injected via the ear vein with a bolus dose of 0.3 mg/kg. Visible and ICG fluorescence images of cortical vessels were simultaneously observed on the display monitor at high resolution. The real-time merging of the visible and fluorescent images corresponded well. Conclusion : Simultaneous visible color and ICG fluorescent imaging of the cortical vessels in the swine brain was satisfactory. Technical improvement and clinical implication are expected.

A Study of Real Time Verification System or Radiation Therapy (방사선치료 위치 실시간 검증시스템에 관한 연구)

  • Kim, Y.J.;Ji, Y.H.;Lee, D.H.;Lee, D.H.;Hong, S.H.
    • Proceedings of the KOSOMBE Conference
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    • v.1997 no.11
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    • pp.164-167
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    • 1997
  • The treatment setup of patients during irradiation is an important aspect in relation to the success of radiotherapy. Imaging with the treatment beam is a widely used method or verification of the radiation field position relative to the target area, prior to or during irradiation. In this paper, Real time digital radiography system was implemented or verification of local error between simulation plan and radiation therapy machine. Portal image can be acquired by CCD camera, image board and pentium PC after therapy Radiation was converted into light by a metal/fluorescent Screen. The resulting image quality is comparable to film, so the imaging system represents a promising alternative to film as a method of verifying patient positioning in radiotherapy. Edge detection and field size measurement were also implemented and detected automatically for verification of treatment position. Field edge was added to the original image or checking the anatomical treatment verification by therapy technicians. By means of therapy efficiency improvement and decrease of Radiation side effects with these techniques, Exact Radiation treatments are expected.

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Visualization of Gene Transfer into Live Cells Using Fluorescent Semiconductor Nanocrystals

  • Kim Jung Kyung;Lim Sun Hee;Lee Yongku;Shin Young Shik;Chung Chanil;Chang Jun Keun;Yoo Jung Yul
    • 한국가시화정보학회:학술대회논문집
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    • 2003.11a
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    • pp.81-82
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    • 2003
  • We have developed the method for the conjugation of biotinylated DNA to streptavidin-coated QDs. QD-DNA conjugates and a high-sensitive fluorescence imaging technique are adopted to visualize gene transport across the membrane of the live cell in real time. Endocytotic cellular uptake of oligonucleotide and electrically-mediated plasmid DNA transfer into the live cell are monitored by a quantitative microscopic imaging system. Long-term kinetic study enables us to reveal the unknown mechanisms and rate-limiting steps of extracellular and intracellular transport of biomolecules. We designed experimental protocols to conjugate the oligonucleotide or the plasmid DNA to commercially available streptavidin-coated QDs. Gel electrophoresis is used to verify the effect of incubation time and the molar ratio of QDs and DNA on the conjugation efficiency. It is possible to fractionate the QD-DNA conjugates according to the DNA concentration and obtain the purified conjugates by a gel extraction technique.

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