• Molecules and Cells
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• 제42권5호
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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.

• Applied Microscopy
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• 제51권
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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.

• Parasites, Hosts and Diseases
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• 제54권2호
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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.

• 한국정보기술학회논문지
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• 제16권11호
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• pp.115-121
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• 2018

근적외선을 이용한 형광 영상은 방사능에 대한 걱정이 없고 수술 중 실시간으로 영상 확인이 가능한 장점이 있다. 따라서 감시림프절 생검에 형광 영상을 이용하는 실험이 활발히 진행되고 있다. 형광 영상장비는 LED, 카메라와 같은 고발열 부품이 사용되어 안정적인 발열 억제 수단으로 수랭 방식의 쿨링 시스템을 사용하고 있다. 형광 영상장비에서 수랭 쿨링 시스템은 큰 부피를 차지하여 장비의 소형화 측면에서는 단점이 된다. 장비의 소형화를 위해 공랭 쿨링 방식을 이용할 경우 발열이 문제가 된다. 본 논문에서는 장비의 소형화를 위해 PWM 제어를 이용한 공랭 방식을 적용하여 실험을 했고, 장비를 장시간 사용해도 문제가 없는 일정한 품질의 형광 영상과 발열 억제 성능을 확인했다.

• 디자인학연구
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• 제19권4호
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• pp.205-208
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• 2006

본 작품은 jitter의 multiplex기능을 이용한 디지털 아트작품으로 두 가지 영상의 cross concept를 이용한 인터렉티브 영상 설치 작품이다. 평면스크린이 아닌 형광등을 이용하여 특수 제작한 스크린을 통해, 형광등에 비춰지는 영상과 그 틈새로 벽에 비춰지는 두 가지 영상을 보여주게 된다. 실시간으로 카메라에 의해 받아들여진 관객의 모습과 제작해 놓은 두 가지 영상은 jitter의 multiplex기능에 의한 합성과정을 거쳐 형광등 스크린에만, 또는 벽에만 비춰지게 된다. 한 영상이지만 관객의 모습은 벽과 형광등을 오가며 관객을 혼란에 빠트리게 된다. 제작한 영상과 실시간으로 입력되는 관객의 두 가지 모습은 상반, 또는 병치의 느낌으로 관객에게 다가가게 되며 그것은 인간의 이중성을 표현하게 되는 동시에 물리적인 존재공간과 우리가 느끼는 시각적 공간을 구분하여 서로 다른 지배 문법이 있다는 것을 표현하게 된다. 퍼스펙티브와 환경에 의해 왜곡되는 영상재현의 과정을 보여주기 위해 형광등으로 제작한 스크린과 cross concept의 영상을 제작 사용하였다.

• Medical Lasers
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• 제9권1호
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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.

• International Journal of Oral Biology
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• 제33권4호
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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.

• Journal of Korean Neurosurgical Society
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• 제58권6호
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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.

• 대한의용생체공학회:학술대회논문집
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• 대한의용생체공학회 1997년도 추계학술대회
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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.

• 한국가시화정보학회:학술대회논문집
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• 한국가시화정보학회 2003년도 추계학술대회 논문집
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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.