• Title/Summary/Keyword: live imaging

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Imaging Single-mRNA Localization and Translation in Live Neurons

  • Lee, Byung Hun;Bae, Seong-Woo;Shim, Jaeyoun Jay;Park, Sung Young;Park, Hye Yoon
    • Molecules and Cells
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    • v.39 no.12
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    • pp.841-846
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    • 2016
  • Local protein synthesis mediates precise spatio-temporal regulation of gene expression for neuronal functions such as long-term plasticity, axon guidance and regeneration. To reveal the underlying mechanisms of local translation, it is crucial to understand mRNA transport, localization and translation in live neurons. Among various techniques for mRNA analysis, fluorescence microscopy has been widely used as the most direct method to study localization of mRNA. Live-cell imaging of single RNA molecules is particularly advantageous to dissect the highly heterogeneous and dynamic nature of messenger ribonucleoprotein (mRNP) complexes in neurons. Here, we review recent advances in the study of mRNA localization and translation in live neurons using novel techniques for single-RNA imaging.

Time-Lapse Live-Cell Imaging Reveals Dual Function of Oseg4, Drosophila WDR35, in Ciliary Protein Trafficking

  • Lee, Nayoung;Park, Jina;Bae, Yong Chul;Lee, Jung Ho;Kim, Chul Hoon;Moon, Seok Jun
    • Molecules and Cells
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    • v.41 no.7
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    • pp.676-683
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    • 2018
  • Cilia are highly specialized antennae-like organelles that extend from the cell surface and act as cell signaling hubs. Intraflagellar transport (IFT) is a specialized form of intracellular protein trafficking that is required for the assembly and maintenance of cilia. Because cilia are so important, mutations in several IFT components lead to human disease. Thus, clarifying the molecular functions of the IFT proteins is a high priority in cilia biology. Live imaging in various species and cellular preparations has proven to be an important technique in both the discovery of IFT and the mechanisms by which it functions. Live imaging of Drosophila cilia, however, has not yet been reported. Here, we have visualized the movement of IFT in Drosophila cilia using time-lapse live imaging for the first time. We found that NOMPB-GFP (IFT88) moves according to distinct parameters depending on the ciliary segment. NOMPB-GFP moves at a similar speed in proximal and distal cilia toward the tip (${\sim}0.45{\mu}m/s$). As it returns to the ciliary base, however, NOMPB-GFP moves at ${\sim}0.12{\mu}m/s$ in distal cilia, accelerating to ${\sim}0.70{\mu}m/s$ in proximal cilia. Furthermore, while live imaging NOMPB-GFP, we observed one of the IFT proteins required for retrograde movement, Oseg4 (WDR35), is also required for anterograde movement in distal cilia. We anticipate our time-lapse live imaging analysis technique in Drosophila cilia will be a good starting point for a more sophisticated analysis of IFT and its molecular mechanisms.

Visualization of chromatin higher-order structures and dynamics in live cells

  • Park, Tae Lim;Lee, YigJi;Cho, Won-Ki
    • BMB Reports
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    • v.54 no.10
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    • pp.489-496
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    • 2021
  • Chromatin has highly organized structures in the nucleus, and these higher-order structures are proposed to regulate gene activities and cellular processes. Sequencing-based techniques, such as Hi-C, and fluorescent in situ hybridization (FISH) have revealed a spatial segregation of active and inactive compartments of chromatin, as well as the non-random positioning of chromosomes in the nucleus, respectively. However, regardless of their efficiency in capturing target genomic sites, these techniques are limited to fixed cells. Since chromatin has dynamic structures, live cell imaging techniques are highlighted for their ability to detect conformational changes in chromatin at a specific time point, or to track various arrangements of chromatin through long-term imaging. Given that the imaging approaches to study live cells are dramatically advanced, we recapitulate methods that are widely used to visualize the dynamics of higher-order chromatin structures.

Adult stem cell lineage tracing and deep tissue imaging

  • Fink, Juergen;Andersson-Rolf, Amanda;Koo, Bon-Kyoung
    • BMB Reports
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    • v.48 no.12
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    • pp.655-667
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    • 2015
  • Lineage tracing is a widely used method for understanding cellular dynamics in multicellular organisms during processes such as development, adult tissue maintenance, injury repair and tumorigenesis. Advances in tracing or tracking methods, from light microscopy-based live cell tracking to fluorescent label-tracing with two-photon microscopy, together with emerging tissue clearing strategies and intravital imaging approaches have enabled scientists to decipher adult stem and progenitor cell properties in various tissues and in a wide variety of biological processes. Although technical advances have enabled time-controlled genetic labeling and simultaneous live imaging, a number of obstacles still need to be overcome. In this review, we aim to provide an in-depth description of the traditional use of lineage tracing as well as current strategies and upcoming new methods of labeling and imaging.

Successful treatment of gastrocnemius muscle rupture in a long-tailed goral rescued from a live trap: a case report

  • Ahn, Sangjin;Choi, Sooyoung;Kim, Jong-Taek
    • Journal of Veterinary Science
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    • v.23 no.3
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    • pp.45.1-45.5
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    • 2022
  • We describe a case of gastrocnemius muscle rupture in a goral that got caught in a live trap set up to capture wild boars. We established a diagnostic method based on clinical signs and imaging examination findings, including magnetic resonance imaging for gastrocnemius muscle rupture with symptoms such as hindlimb lameness and plantigrade posture of unknown cause in wild animals. Although this condition is rare, this report will help veterinarians diagnose several diseases of the musculoskeletal and nervous systems in wild animals. This is the first report of successful diagnosis and treatment of gastrocnemius muscle rupture in gorals in Korea.

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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Development of a virtual studio system for live broadcasting of election results: VdreamSet

  • Ko, Hee-Dong;Kim, Hyun-Suk;Kim, Lae-Hyun;Ahn, Jae-Hong;Park, Kyung-Dong
    • Proceedings of the Korean Society of Broadcast Engineers Conference
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    • 1999.06a
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    • pp.213.1-218
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    • 1999
  • A virtual studio is a new video production environment using interactive computer graphics and imaging media technology. The traditional chroma-keying with two-dimensional background is replaced by an advanced keying method with a dynamic computer-generated, three-dimensional background. We have developed a virtual studio system that is practical to use in the real production environment. It has not only essential features that are common among various commercial virtual studio systems, but also unique feature that help the producer to construct virtual studio sets and scenarios efficiently such as span graph, robust backup controller, and 3 dimensional character generator supporting all languages. Our virtual studio system was used in live broadcasting and proved that the system was practical enough. In this paper, we will introduce the structure and the major features of our system, called VdreamSet, and application examples to broadcasting.

Live Electrooptic Imaging Camera for Real-Time Visual Accesses to Electric Waves in GHz Range

  • Tsuchiya, Masahiro;Shiozawa, Takahiro
    • Journal of electromagnetic engineering and science
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    • v.11 no.4
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    • pp.290-297
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    • 2011
  • Recent progresses in the live electrooptic imaging (LEI) technique are reviewed with emphasis on its functionality of real-time visual accesses to traveling electric waves in the GHz range. Together with the principles, configurations, and procedures for the visual observation experiments by an LEI camera system, the following results are described as examples indicating the wide application ranges of the technique; Ku-band waves on arrayed planar antennas, waves on a Gb/s-class digital circuit, W-band waves traveling both in slab-waveguide modes and aerially, backward-traveling wave along composite right/left-handed transmission line, and, waves in monolithic microwave integrated circuit module case.