• Title/Summary/Keyword: Cellular imaging

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Use of DNA-Specific Anthraquinone Dyes to Directly Reveal Cytoplasmic and Nuclear Boundaries in Live and Fixed Cells

  • Edward, Roy
    • Molecules and Cells
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    • v.27 no.4
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    • pp.391-396
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    • 2009
  • Image-based, high-content screening assays demand solutions for image segmentation and cellular compartment encoding to track critical events - for example those reported by GFP fusions within mitosis, signalling pathways and protein translocations. To meet this need, a series of nuclear/cytoplasmic discriminating probes have been developed: DRAQ5$^{TM}$ and CyTRAK Orange$^{TM}$. These are spectrally compatible with GFP reporters offering new solutions in imaging and cytometry. At their most fundamental they provide a convenient fluorescent emission signature which is spectrally separated from the commonly used reporter proteins (e.g. eGFP, YFP, mRFP) and fluorescent tags such as Alexafluor 488, fluorescein and Cy2. Additionally, they do not excite in the UV and thus avoid the complications of compound UV-autofluorescence in drug discovery whilst limiting the impact of background sample autofluorescence. They provide a convenient means of stoichiometrically labelling cell nuclei in live cells without the aid of DMSO and can equally be used for fixed cells. Further developments have permitted the simultaneous and differential labelling of both nuclear and cytoplasmic compartments in live and fixed cells to clearly render the precise location of cell boundaries which may be beneficial for quantitative expression measurements, cell-cell interactions and most recently compound in vitro toxicology testing.

Functional Dissection of Glutamatergic and GABAergic Neurons in the Bed Nucleus of the Stria Terminalis

  • Kim, Seong-Rae;Kim, Sung-Yon
    • Molecules and Cells
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    • v.44 no.2
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    • pp.63-67
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    • 2021
  • The bed nucleus of the stria terminalis (BNST)-a key part of the extended amygdala-has been implicated in the regulation of diverse behavioral states, ranging from anxiety and reward processing to feeding behavior. Among the host of distinct types of neurons within the BNST, recent investigations employing cell type- and projection-specific circuit dissection techniques (such as optogenetics, chemogenetics, deep-brain calcium imaging, and the genetic and viral methods for targeting specific types of cells) have highlighted the key roles of glutamatergic and GABAergic neurons and their axonal projections. As anticipated from their primary roles in excitatory and inhibitory neurotransmission, these studies established that the glutamatergic and GABAergic subpopulations of the BNST oppositely regulate diverse behavioral states. At the same time, these studies have also revealed unexpected functional specificity and heterogeneity within each subpopulation. In this Minireview, we introduce the body of studies that investigated the function of glutamatergic and GABAergic BNST neurons and their circuits. We also discuss unresolved questions and future directions for a more complete understanding of the cellular diversity and functional heterogeneity within the BNST.

Functional Imaging of the Multidrug Resistance In Vivo (기능적 영상술을 이용한 다약제 내성의 체내 진단)

  • Lee, Jea-Tae
    • 대한핵의학회:학술대회논문집
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    • 2001.05a
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    • pp.66-75
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    • 2001
  • Although diverse mechanisms are involved in multidrug resistance for chemotherapeutic drugs, the development of cellular P-glycoprotein(Pgp) and multidrug-resistance associated protein (MRP) are important factors in the chemotherapy failure to cancer. Various detection assays provide information about the presence of drug efflux pumps at the mRNA and protein levels. However these methods do not yield information about dynamic function of Pgp and MRP un vivo. Single photon emission tomography (SPECT) and positron emission tomography (PET) are available for the detection of Pgp and MRP-mediated transport. $^{99m}Tc$-sestaMIBl and other $^{99m}Tc$-radiopharmaceuticals are substrates for Pgp and MRP, and have been used in clinical studies for tumor imaging, and to visualize blockade of Pgp-mediated transport after modulation of Pgp pump. Colchicine, verapamil and daunorubicin labeled with $^{11}C$ have been evaluated for the quantification of Pgp-mediated transport with PET in vivo and reported to be feasible substrates with which to image Pgp function in tumors. Leukotrienes are specific substrates for MRP and N-$[^{11}C]$acetyl-leukotriene E4 provides an opportunity to study MRP function non-invasively in vivo. Results obtained from recent publications are reviewed to confirm the feasibility of using SPECT and PET to study the functionality of MDR transporters in vivo.

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Cytoprotective Constituents of the Stem Barks of Fraxinus rhynchophylla on Mouse Hippocampal HT22 Cells and Their Antioxidative Activity (물푸레나무 수피의 생쥐 해마 유래 HT22 세포 보호와 항산화 활성 물질)

  • Jeong, Gil-Saeng;Yoon, Kwon-Ha;Kim, Hyun-Chul;Oh, Seung-Hwan;Kim, Myong-Jung;Kang, Dae-Gill;Lee, Ho-Sub;Kim, Youn-Chul
    • Korean Journal of Pharmacognosy
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    • v.38 no.3 s.150
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    • pp.287-290
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    • 2007
  • Phytochemical investigation of the MeOH extract of the dried stem barks of Fraxinus rhynchophylla Hance (Oleaceae), as guided by cytoprotective activity against tert-butyl hydroperoxide (t-BHP)-induced cell injury in mouse hippocampal HT22 cells, furnished two coumarins, esculetin (1) and fraxetin (2). Compounds 1 and 2 had the significant cytoprotective effects on t-BHP-induced cellular oxidative injury in HT22 cells. Furthermore, compounds 1 and 2 showed potent DPPH radical scavenging effect, exhibiting $IC_{50}$ values of 14.68 and 9.64 ${\mu}M$, respectively.

A comparison of photosynthesis measurements by O2 evolution, 14C assimilation, and variable chlorophyll fluorescence during light acclimatization of the diatom Coscinodiscus granii

  • Trampe, Erik;Hansen, Per Juel;Kuhl, Michael
    • ALGAE
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    • v.30 no.2
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    • pp.103-119
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    • 2015
  • Photosynthetic rates of the large centric diatom Coscinodiscus granii were measured by means of multicolor variable chlorophyll fluorescence imaging, single cell $^{14}C$ assimilation, and optical $O_2$ sensor measurements during light acclimatization of cultures grown at five different irradiances: 50, 150, 235, 332, and $450{\mu}mol$ photons $m^{-2}\;s^{-1}$. Photo-acclimatization was evident from changes of cellular chlorophyll a content, growth rates, and light response curves. Each of the applied methods evaluates different parts and reactions in the photosynthetic apparatus, which makes a direct quantitative comparison of rates difficult, although a different degree of correlation were found between all three methods. However, when used in combination, they provide information about the internal relationship of photosynthetic pathways as well as the variation in photosynthetic capacity between individual cells within a single algal culture.

Analysis of SAR in a Human Head for a Cellular Phone (셀룰라 휴대폰에 의한 인체 두부의 SAR 해석)

  • 이애경;최형도;김진석
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.9 no.6
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    • pp.776-787
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    • 1998
  • This paper analyzes the local specific absorption rates (SAR's) averaged over 1 g and 10 g in a human head model in contact with a mobile phone operating at 835 MHz. The used numerical method is a total field finite-difference time-domain (FDTD) technique. The phone was simulated with a conducting box, a plastic case, and a whip antennal composed of a monopole and a helix. The discrete human model of the spatial resolution 3 mm is based on Magnetic Resonance Imaging (MRI), computerized tomography (CT) and anatomical images. The near field and far field and far field patterns were analyzed for extended and retracted phone. The two methods to take the volumes of the weights, 1 g or 10 g in tissue are proposed and compared to offer a reproductive technique for SAR estimations.

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Development of line-scanning two-photon microscopy based on spatial and temporal focusing for tryptophan based auto fluorescence imaging (고속 트립토판 자가형광 이미징을 위한 시공간적 집중 기반의 라인 스캐닝 이광자 현미경 개발)

  • Lee, Jun Ho;Nam, Hyo Seok;Kim, Ki Hean
    • Journal of the Korean Society of Visualization
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    • v.11 no.2
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    • pp.41-45
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    • 2013
  • Two-photon microscopy (TPM) is minimally-invasive 3D fluorescence microscopy based on nonlinear excitation, and TPM can visualize cellular structures based on auto-fluorescence. Line-scanning TPM is one of high-speed TPM methods without sacrificing the image resolution by using spatial and temporal focusing. In this paper, we developed line-scanning TPM based on spatial and temporal focusing for auto-fluorescence imaging by exciting the tryptophan. Laser source for this system was an optical parametric oscillator (OPO) and it made near 570 nm femtosecond pulse laser. It had 200fs pulse width and 1.72 nm bandwidth, so that the achievable depth resolution was 2.41um and field of view (FOV) is 10.8um. From the characterization, our system has 3.0 um depth resolution and 12.3 um FOV. We visualized fixed leukocyte cell sample and compared with point scanning system.

Identification of Novel Regulators of Apoptosis Using a High-Throughput Cell-based Screen

  • Park, Kyung Mi;Kang, Eunju;Jeon, Yeo-Jin;Kim, Nayoung;Kim, Nam-Soon;Yoo, Hyang-Sook;Yeom, Young Il;Kim, Soo Jung
    • Molecules and Cells
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    • v.23 no.2
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    • pp.170-174
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    • 2007
  • High-throughput subcellular imaging is a powerful tool for investigating the function of genes. In order to identify novel regulators of apoptosis we transiently transfected HeLa cells with 938 hypothetical genes of unknown function, and captured their nuclear images with an automated fluorescence microscope. We selected genes that induced greater than 3-fold increase in the percentage of apoptotic nuclei compared with vector-transfected cells. The full-length genes C10orf61, MGC 26717, and FLJ13855 were identified as candidate proapoptotic genes, and their apoptotic effects were confirmed by DNA fragmentation ELISAs and Western blotting for caspase-7 and PARP. We conclude that a subcellular image-based apoptotic screen is useful for identifying genes with proapoptotic activity.

A Study on the PET/CT Fusion Imaging (PET/CT 결합영상진단 검사에 관한 연구)

  • Kim, Jong Gyu
    • Korean Journal of Clinical Laboratory Science
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    • v.36 no.2
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    • pp.193-198
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    • 2004
  • PET/CT combines the functional information from a positron emission tomography (PET) exam with the anatomical information from a computed tomography (CT) exam into one single exam. A CT scan uses a combination of x-rays and computers to give the radiologist a non-invasive way to see inside your body. One advantage of CT is its ability to rapidly acquire two-dimensional pictures of your anatomy. Using a computer these 2-D images can be presented in 3-D for in-depth clinical evaluation. A PET scan detects changes in the cellular function - how your cells are utilizing nutrients like sugar and oxygen. Since these functional changes take place before physical changes occur, PET can provide information that enables your physician to make an early diagnosis. The PET exam pinpoints metabolic activity in cells and the CT exam provides an anatomical reference. When these two scans are fused together, your physician can view metabolic changes in the proper anatomical context of your body. PET/CT offers significant advantages including more accurate localization of functional abnormalities, and the distinction of pathological from normal physiological uptake, and improvements in monitoring treatment. A PET/CT scan allows physicians to measure the body's abnormal molecular cell activity to detect cancer (such as breast cancer, lung cancer, colorectal cancer, lymphoma, melanoma and other skin cancers), brain disorders (such as Alzheimer's disease, Parkinson's disease, and epilepsy), and heart disease (such as coronary artery disease).

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Spatiotemporal characteristics of atrial $Ca^{2+}$ sparks: evidence from two-dimensional rapid confocal imaging

  • Woo, Sun-Hee;Lars Cleemann;Martin Morad
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.25-25
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    • 2003
  • Atrial myocytes have two functionally separate $Ca^{2+}$ release sites: those in peripheral sarcoplasmic reticulum (SR) adjacent to the $Ca^{2+}$ channels of surface membrane and those in central SR not associated with $Ca^{2+}$ channels. Study on the spatio-temporal properties of focal $Ca^{2+}$ releases (“sparks”) occurring spontaneously in central and peripheral sites of voltage-clamped rat atrial myocytes, using rapid two-dimensional (2-D) confocal $Ca^{2+}$ imaging revealed that peripheral and central sparks were similar in size and release time (~300,000 $Ca^{2+}$ ions for=12 ms), but significantly larger and longer than ventricular sparks. Both sites were resistant to Cd$^{2+}$ and inhibited by ryanodine. Peripheral sparks were brighter and flattened against surface membrane, had ~5-fold higher frequency, ~2 times faster diffusion coefficient, and dissipated abruptly. Central sparks, in contrast, occurred less frequently, were elongated along the cellular longitudinal axis, and dissipated slowly. Compound sparks (composed of 2-5 unitary focal releases) aligned longitudinally, occurred more frequently at the center.at the center.

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