• 제목/요약/키워드: Cellular imaging

검색결과 180건 처리시간 0.024초

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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    • 제41권7호
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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.

MicroSPECT and MicroPET Imaging of Small Animals for Drug Development

  • Jang, Beom-Su
    • Toxicological Research
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    • 제29권1호
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    • pp.1-6
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    • 2013
  • The process of drug discovery and development requires substantial resources and time. The drug industry has tried to reduce costs by conducting appropriate animal studies together with molecular biological and genetic analyses. Basic science research has been limited to in vitro studies of cellular processes and ex vivo tissue examination using suitable animal models of disease. However, in the past two decades new technologies have been developed that permit the imaging of live animals using radiotracer emission, X-rays, magnetic resonance signals, fluorescence, and bioluminescence. The main objective of this review is to provide an overview of small animal molecular imaging, with a focus on nuclear imaging (single photon emission computed tomography and positron emission tomography). These technologies permit visualization of toxicodynamics as well as toxicity to specific organs by directly monitoring drug accumulation and assessing physiological and/or molecular alterations. Nuclear imaging technology has great potential for improving the efficiency of the drug development process.

Adult stem cell lineage tracing and deep tissue imaging

  • Fink, Juergen;Andersson-Rolf, Amanda;Koo, Bon-Kyoung
    • BMB Reports
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    • 제48권12호
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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.

혈관신생 분자핵의학 영상 (Molecular Nuclear imaging of Angiogenesis)

  • 이경한
    • 대한핵의학회지
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    • 제38권2호
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    • pp.171-174
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    • 2004
  • Angiogenesis, the formation of new capillaries from existing vessels, increases oxygenation and nutrient supply to ischemic tissue and allows tumor growth and metastasis. As such, angiogenesis targeting provides a novel approach for cancer treatment with easier drug delivery and less drug resistance. Therapeutic anti-angiogenesis has shown impressive effects in animal tumor models and are now entering clinical trials. However, the successful clinical introduction of this new therapeutic approach requires diagnostic tools that can reliably measure angiogenesis in a noninvasive and repetitive manner. Molecular imaging is emerging as an exciting new discipline that deals with imaging of disease on a cellular or genetic level. Angiogenesis imaging is an important area for molecular imaging research, and the use of radiotracers offers a particularly promising technique for its development. While current perfusion and metabolism radiotracers can provide useful information related to tissue vascularity, recent endeavors are focused on the development of novel radioprobes that specifically and directly target angiogenic vessels. Presently available proges include RGD sequence containing peptides that target ${\alpha}_v\;{\beta}_3$ integrin, endothelial growth factors such as VEGF or FGF, metalloptoteinase inhibitors, and specific antiangiogenic drugs. It is now clear that nuclear medicine techniques have a remarkable potential for angiogenesis imaging, and efforts are currently continuing to develop new radioprobes with superior imaging properties. With future identification of novel targets, design of better probes, and improvements in instrumentation, radiotracer angiogenesis imaging promises to play an increasingly important role in the diagnostic evaluation and treatment of cancer and other angiogenesis related diseases.

Simultaneous Evaluation of Cellular Vitality and Drug Penetration in Multicellular Layers of Human Cancer Cells

  • Al-Abd Ahmed Mohammed;Lee Joo-Ho;Kuh Hyo-Jeong
    • Journal of Pharmaceutical Investigation
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    • 제36권5호
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    • pp.309-314
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    • 2006
  • The multicellular layers(MCL) of human cancer cells is a three dimensional(3D) in vitro model for human solid tumors which has been used primarily for the assessment of avascular penetration of anti-cancer drugs. For anti-cancer drugs with penetration problem, MCL represents a good experimental model that can provide clinically relevant data. Calcein-AM is a fluorescent dye that demonstrates the cellular vitality in a graded manner in cancer cell culture system. In the present study, we evaluated the use of calcein-AM for determination of anti-proliferative activity of anti-cancer agents in MCL model of DLD-1 human colorectal cancer cells. Optical sectioning of confocal imaging was compromised with photonic attenuation and penetration barrier in the deep layers of MCL. By contrast, fluorescent measurement on the cryo-sections provided a feasible alternative. Cold pre-incubation did not enhance the calcein-AM distribution to a significant degree in MCL of DLD-1 cells. However, the simultaneous determination of drug penetration and cellular vitality appeared to be possible in drug treated MCL. In conclusion, these data suggest that calcein-AM can be used for the simultaneous determination of drug-induced anti-proliferative effect and drug penetration in MCL model.

Characterizing Organelles in Live Stem Cells Using Label-Free Optical Diffraction Tomography

  • Kim, Youngkyu;Kim, Tae-Keun;Shin, Yeonhee;Tak, Eunyoung;Song, Gi-Won;Oh, Yeon-Mok;Kim, Jun Ki;Pack, Chan-Gi
    • Molecules and Cells
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    • 제44권11호
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    • pp.851-860
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    • 2021
  • Label-free optical diffraction tomography (ODT), an imaging technology that does not require fluorescent labeling or other pre-processing, can overcome the limitations of conventional cell imaging technologies, such as fluorescence and electron microscopy. In this study, we used ODT to characterize the cellular organelles of three different stem cells-namely, human liver derived stem cell, human umbilical cord matrix derived mesenchymal stem cell, and human induced pluripotent stem cell-based on their refractive index and volume of organelles. The physical property of each stem cell was compared with that of fibroblast. Based on our findings, the characteristic physical properties of specific stem cells can be quantitatively distinguished based on their refractive index and volume of cellular organelles. Altogether, the method employed herein could aid in the distinction of living stem cells from normal cells without the use of fluorescence or specific biomarkers.

Development of Two-Component Nanorod Complex for Dual-Fluorescence Imaging and siRNA Delivery

  • Choi, Jin-Ha;Oh, Byung-Keun
    • Journal of Microbiology and Biotechnology
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    • 제24권9호
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    • pp.1291-1299
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    • 2014
  • Recently, multifunctional nanomaterials have been developed as nanotherapeutic agents for cellular imaging and targeted cancer treatment because of their ease of synthesis and low cytotoxicity. In this study, we developed a multifunctional, two-component nanorod consisting of gold (Au) and nickel (Ni) blocks that enables dual-fluorescence imaging and the targeted delivery of small interfering RNA (siRNA) to improve cancer treatment. Fluorescein isothiocyanate-labeled luteinizing hormone-releasing hormone (LHRH) peptides were attached to the surface of a Ni block via a histidine-tagged LHRH interaction to specifically bind to a breast cancer cell line, MCF-7. The Au block was modified with TAMRA-labeled thiolated siRNA in order to knock down the vascular endothelial growth factor protein to inhibit cancer growth. These two-component nanorods actively targeted and internalized into MCF-7 cells to induce apoptosis through RNA interference. This study demonstrates the feasibility of using two-component nanorods as a potential theranostic in breast cancer treatment, with capabilities in dual imaging and targeted gene delivery.

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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    • 제39권12호
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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.

Comparison of D-[18F]FMAU and L-[18F]FMAU as PET Imaging Agents for HSV1-TK Gene Expression

  • Moon, Byung-Seok;Jo, Nam-Hyun;Lee, Kyo-Chul;El-Gamal, Mohammed I.;An, Gwang-Il;Hong, Su-Hee;Choi, Tae-Hyun;Choi, Won-Kyoung;Park, Jin-Hun;Cho, Jung-Hyuck;Cheon, Gi-Jeong;Oh, Chang-Hyun
    • Bulletin of the Korean Chemical Society
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    • 제31권11호
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    • pp.3309-3312
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    • 2010
  • D-[$^{18}F$]FMAU and L-[$^{18}F$]FMAU are F-18 labeled nucleoside analogue which have been efficiently synthesized in order to be a PET imaging probe. D-[$^{18}F$]FMAU and L-[$^{18}F$]FMAU were compared as PET imaging agents using HSV1-TK gene expressing tumor-bearing mice. Their cellular uptake profiles were also compared using MCA and MCA-TK cell lines. D-[$^{18}F$]FMAU demonstrated higher cellular uptake and higher accumulation in MCA-TK tumor regions than L-[$^{18}F$]FMAU. On the other hand, L-[$^{18}F$]FMAU showed higher MCA-TK/MCA ratio of %ID/g than that of D-[$^{18}F$]FMAU. L-[$^{18}F$]FMAU can be utilized as a good candidate for HSV1-TK PET imaging. It can be used for antiviral drug evaluation.