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Live cell imaging

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https://www.readbyqxmd.com/read/29453903/tracking-the-dynamic-folding-and-unfolding-of-rna-g-quadruplexes-in-live-cells
#1
Xiu-Cai Chen, Shuo-Bin Chen, Jing Dai, Jia-Hao Yuan, Tian-Miao Ou, Zhi-Shu Huang, Jia-Heng Tan
Due to the absence of methods for tracking RNA G-quadruplex dynamics, especially the folding and unfolding of this attractive structure in live cells, understanding of the biological roles of RNA G-quadruplexes is so far limited. Here we reported a new red-emitting fluorescent probe, QUMA-1, for a selective, continuous and real-time visualization of RNA G-quadruplexes in live cells. The applications of QUMA-1 in several previously intractable applications, including live-cell imaging of the dynamic folding, unfolding and movement of RNA G-quadruplexes and visualizing the unwinding of RNA G-quadruplexes by RNA helicase have been demonstrated...
February 17, 2018: Angewandte Chemie
https://www.readbyqxmd.com/read/29453535/toxicity-of-metal-oxide-nanoparticles
#2
Koyeli Girigoswami
In the recent times, nanomaterials are used in many sectors of science, medicine and industry, without revealing its toxic effects. Thus, it is in urgent need for exploring the toxicity along with the application of such useful nanomaterials. Nanomaterials are categorized with a particle size of 1-100 nm. They have gained increasing attention because of their novel properties, including a large specific surface area and high reaction activity. The various fundamental and practical applications of nanomaterials include drug delivery, cell imaging, and cancer therapy...
2018: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/29453143/structure-and-physico-mechanical-properties-of-low-temperature-plasma-treated-electrospun-nanofibrous-scaffolds-examined-with-atomic-force-microscopy
#3
Adrian Chlanda, Ewa Kijeńska, Chiara Rinoldi, Michał Tarnowski, Tadeusz Wierzchoń, Wojciech Swieszkowski
Electrospun nanofibrous scaffolds are willingly used in tissue engineering applications due to their tunable mechanical, chemical and physical properties. Additionally, their complex openworked architecture is similar to the native extracellular matrix of living tissue. After implantation such scaffolds should provide sufficient mechanical support for cells. Moreover, it is of crucial importance to ensure sterility and hydrophilicity of the scaffold. For this purpose, a low temperature surface plasma treatment can be applied...
February 8, 2018: Micron: the International Research and Review Journal for Microscopy
https://www.readbyqxmd.com/read/29453096/design-of-mitochondria-targeted-colorimetric-and-ratiometric-fluorescent-probes-for-rapid-detection-of-so-2-derivatives-in-living-cells
#4
Yutao Yang, Tingting Zhou, Bozan Bai, Caixia Yin, Wenzhi Xu, Wei Li
Two mitochondria-targeted colorimetric and ratiometric fluorescent probes for SO 2 derivatives were constructed based on the SO 2 derivatives-triggered Michael addition reaction. The probes exhibit high specificity toward HSO 3 - /SO 3 2- by interrupting their conjugation system resulting in a large ratiometric blue shift of 46-121nm in their emission spectrum. The two well-resolved emission bands can ensure accurate detection of HSO 3 - . The detection limits were calculated to be 1.09 and 1.35μM. Importantly, probe 1 and probe 2 were successfully used to fluorescence ratiometric imaging of endogenous HSO 3 - in BT-474 cells...
February 13, 2018: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/29452287/dental-pulp-stem-cell-derived-chondrogenic-cells-demonstrate-differential-cell-motility-in-type-i-and-type-ii-collagen-hydrogels
#5
Li Yao, Nikol Flynn
BACKGROUND CONTEXT: Advances in the development of biomaterials and stem cell therapy provide a promising approach to regenerating degenerated discs. The normal nucleus pulposus (NP) cells exhibit the similar phenotype as chondrocytes. Because dental pulp stem cells (DPSCs) can be differentiated into chondrogenic cells, the DPSCs and DPSCs-derived chondrogenic cells encapsulated in type I and type II collagen hydrogels can potentially be transplanted into degenerated nucleus pulposus (NP) to repair damaged tissue...
February 13, 2018: Spine Journal: Official Journal of the North American Spine Society
https://www.readbyqxmd.com/read/29451726/single-organelle-dynamics-linked-to-3d-structure-by-correlative-live-cell-3d-electron-microscopy
#6
Job Fermie, Nalan Liv, Corlinda Ten Brink, Elly G van Donselaar, Wally H Müller, Nicole L Schieber, Yannick Schwab, Hans C Gerritsen, Judith Klumperman
Live-cell correlative light-electron microscopy (live-cell-CLEM) integrates live movies with the corresponding EM image, but a major challenge is to relate the dynamic characteristics of single organelles to their 3D ultrastructure. Here we introduce focused ion beam scanning electron microscopy (FIB-SEM) in a modular live-cell CLEM pipeline for single organelle CLEM. We transfected cells with LAMP-1-GFP, analyzed the dynamics of individual GFP-positive spots, and correlated these to their corresponding fine-architecture and immediate cellular environment...
February 16, 2018: Traffic
https://www.readbyqxmd.com/read/29451281/fluorescent-antitumor-titanium-iv-salen-complexes-for-cell-imaging
#7
Avia Tzubery, Naomi Melamed-Book, Edit Y Tshuva
Two differently substituted fluorescent salen Ti(iv) complexes were developed. One was inactive on human cancer cells, whereas the other showed high cytotoxicity. Based on live cell imaging, both complexes penetrated the cell, but were not detected in the nuclei. Moreover, the inactive complex was trapped in endocytic vesicles, whereas the active complex accumulated in the perinuclear region and inflected phototoxicity upon continuous irradiation.
February 16, 2018: Dalton Transactions: An International Journal of Inorganic Chemistry
https://www.readbyqxmd.com/read/29449902/sers-fluorescence-bimodal-nanoprobes-for-in-vitro-imaging-of-fatty-acid-responsive-receptor-gpr120
#8
Lifu Xiao, Abdul K Parchur, Timothy A Gilbertson, Anhong Zhou
G-protein-coupled receptor 120 (GPR120), as a member of the rhodopsin family of G-protein-coupled receptors, has been shown to function as a sensor for dietary fat in the gustatory and digestive systems. Its specific role in the chemoreception of fatty acids, which is thought to be crucial in understanding the mechanism surrounding the control of fat intake and, accordingly, in the treatment of obesity, remains unclear. Here we report a novel surface-enhanced Raman spectroscopy (SERS)-fluorescence bimodal microscopic technique for detection and imaging of GPR120 in single living cells...
2018: Analytical Methods: Advancing Methods and Applications
https://www.readbyqxmd.com/read/29448520/synthesis-of-fluorophore-decorated-single-walled-carbon-nanotubes-for-in-vitro-cytocompatibility
#9
Adhigan Murali, Neelamegan Haridharan, Pombala Suresh Babu, Sivalingam Jayaveni, Sellamuthu N Jaisankar
We report rhodamine based fluorophore derivative for the stable dispersion of single-walled carbon nanotubes (SWCNTs), which can afford better fluorescent label to carbon nanotubes (CNTs). The nanotubes-fluorophore conjugates are helpful in achieving stable dispersion in polar and non-polar solvents with intense fluorescence. The product was characterized through NMR, Mass spectrometry, Raman, XPS, SEM, AFM and Fluorescence measurements. The formation of SWCNT-g- Rhodamine was confirmed by the presence of D and G bands in Raman spectrum...
February 1, 2018: Journal of Nanoscience and Nanotechnology
https://www.readbyqxmd.com/read/29446757/3d-bioprinting-mesenchymal-stem-cell-laden-construct-with-core-shell-nanospheres-for-cartilage-tissue-engineering
#10
Wei Zhu, Haitao Cui, Benchaa Boualam, Fahed Masood, Erin Flynn, Raj Rao, Zhiyong Zhang, Lijie Grace Zhang
Cartilage tissue is prone to degradation and has little capacity for self-healing due to its avascularity. Tissue engineering, which provides artificial scaffolds to repair injured tissues, is a novel and promising strategy for cartilage repair. 3D bioprinting offers even greater potential for repairing degenerative tissue by simultaneously integrating living cells, biomaterials, and biological cues to provide a customized scaffold. With regard to cell selection, mesenchymal stem cells (MSCs) hold great capacity for differentiating into a variety of cell types, including chondrocytes, and could therefore be utilized as a cartilage cell source in 3D bioprinting...
February 15, 2018: Nanotechnology
https://www.readbyqxmd.com/read/29446627/ultrabright-and-fluorogenic-probes-for-multicolor-imaging-and-tracking-of-lipid-droplets-in-cells-and-tissues
#11
Mayeul Collot, Tkhe-Kyong Fam, Pichandi Ashokkumar, Orestis Faklaris, Thierry Galli, Lydia Danglot, Andrey S Klymchenko
Lipid droplets (LDs) are intracellular lipid-rich organelles that regulate the storage of neutral lipids and were recently found to be involved in many physiological processes, metabolic disorders as well as diseases including obesity, diabetes and cancers. Herein we present a family of new fluorogenic merocyanine fluorophores based on an indolenine moiety and a dioxaborine barbiturate derivative. These so-called StatoMerocyanines (SMCy) span their fluorescence from yellow to the near infrared (NIR) in oil with an impressive fluorescence enhancement compared to aqueous media...
February 15, 2018: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/29445211/bronchoscopic-fibered-confocal-fluorescence-microscopy-for-longitudinal-in-vivo-assessment-of-pulmonary-fungal-infections-in-free-breathing-mice
#12
Liesbeth Vanherp, Jennifer Poelmans, Amy Hillen, Kristof Govaerts, Sarah Belderbos, Tinne Buelens, Katrien Lagrou, Uwe Himmelreich, Greetje Vande Velde
Respiratory diseases, such as pulmonary infections, are an important cause of morbidity and mortality worldwide. Preclinical studies often require invasive techniques to evaluate the extent of infection. Fibered confocal fluorescence microscopy (FCFM) is an emerging optical imaging technique that allows for real-time detection of fluorescently labeled cells within live animals, thereby bridging the gap between in vivo whole-body imaging methods and traditional histological examinations. Previously, the use of FCFM in preclinical lung research was limited to endpoint observations due to the invasive procedures required to access lungs...
February 14, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29444710/a-novel-in-vitro-model-for-the-assessment-of-postnatal-myonuclear-accretion
#13
Anita Kneppers, Lex Verdijk, Chiel de Theije, Mark Corten, Ellis Gielen, Luc van Loon, Annemie Schols, Ramon Langen
BACKGROUND: Due to the post-mitotic nature of myonuclei, postnatal myogenesis is essential for skeletal muscle growth, repair, and regeneration. This process is facilitated by satellite cells through proliferation, differentiation, and subsequent fusion with a pre-existing muscle fiber (i.e., myonuclear accretion). Current knowledge of myogenesis is primarily based on the in vitro formation of syncytia from myoblasts, which represents aspects of developmental myogenesis, but may incompletely portray postnatal myogenesis...
February 14, 2018: Skeletal Muscle
https://www.readbyqxmd.com/read/29443498/spectrally-resolved-and-functional-super-resolution-microscopy-via-ultrahigh-throughput-single-molecule-spectroscopy
#14
Rui Yan, Seonah Moon, Samuel J Kenny, Ke Xu
As an elegant integration of the spatial and temporal dimensions of single-molecule fluorescence, single-molecule localization microscopy (SMLM) overcomes the diffraction-limited resolution barrier of optical microscopy by localizing single molecules that stochastically switch between fluorescent and dark states over time. While this type of super-resolution microscopy (SRM) technique readily achieves remarkable spatial resolutions of ∼10 nm, it typically provides no spectral information. Meanwhile, current scanning-based single-location approaches for mapping the positions and spectra of single molecules are limited by low throughput and are difficult to apply to densely labeled (bio)samples...
February 14, 2018: Accounts of Chemical Research
https://www.readbyqxmd.com/read/29443110/imaging-approaches-to-assessments-of-toxicological-oxidative-stress-using-genetically-encoded-fluorogenic-sensors
#15
Elizabeth M Corteselli, James M Samet, Eugene A Gibbs-Flournoy
While oxidative stress is a commonly cited toxicological mechanism, conventional methods to study it suffer from a number of shortcomings, including destruction of the sample, introduction of potential artifacts, and a lack of specificity for the reactive species involved. Thus, there is a current need in the field of toxicology for non-destructive, sensitive, and specific methods that can be used to observe and quantify intracellular redox perturbations, more commonly referred to as oxidative stress. Here, we present a method for the use of two genetically-encoded fluorogenic sensors, roGFP2 and HyPer, to be used in live-cell imaging studies to observe xenobiotic-induced oxidative responses...
February 7, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29443100/single-cell-resolution-fluorescence-live-imaging-of-drosophila-circadian-clocks-in-larval-brain-culture
#16
Virginie Sabado, Emi Nagoshi
The circadian pacemaker circuit orchestrates rhythmic behavioral and physiological outputs coordinated with environmental cues, such as day/night cycles. The molecular clock within each pacemaker neuron generates circadian rhythms in gene expression, which underlie the rhythmic neuronal functions essential to the operation of the circuit. Investigation of the properties of the individual molecular oscillators in different subclasses of pacemaker neurons and their interaction with neuronal signaling yields a better understanding of the circadian pacemaker circuit...
January 19, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29443098/a-novel-in-vitro-live-imaging-assay-of-astrocyte-mediated-phagocytosis-using-ph-indicator-conjugated-synaptosomes
#17
Youkyeong Gloria Byun, Won-Suk Chung
Astrocytes are the major cell type in the brain and directly contact synapses and blood vessels. Although microglial cells have been considered the major immune cells and only phagocytes in the brain, recent studies have shown that astrocytes also participate in various phagocytic processes, such as developmental synapse elimination and clearance of amyloid beta plaques in Alzheimer's disease (AD). Despite these findings, the efficiency of astrocyte engulfment and degradation of their targets is unclear compared with that of microglia...
February 5, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29443094/in-vivo-imaging-of-muscle-tendon-morphogenesis-in-drosophila-pupae
#18
Sandra B Lemke, Frank Schnorrer
Muscles together with tendons and the skeleton enable animals including humans to move their body parts. Muscle morphogenesis is highly conserved from animals to humans. Therefore, the powerful Drosophila model system can be used to study concepts of muscle-tendon development that can also be applied to human muscle biology. Here, we describe in detail how morphogenesis of the adult muscle-tendon system can be easily imaged in living, developing Drosophila pupae. Hence, the method allows investigating proteins, cells and tissues in their physiological environment...
February 6, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29443052/intravital-microscopy-of-tumor-associated-vasculature-using-advanced-dorsal-skinfold-window-chambers-on-transgenic-fluorescent-mice
#19
Ann L B Seynhaeve, Timo L M Ten Hagen
Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static information and is often not sufficient for a correct interpretation. Intravital evaluation, in which a process is followed in time, provides extra and often unexpected information. With the creation of transgenic animals expressing cell-specific markers and live cell tracers, improvements to imaging equipment, and the development of several imaging chambers, intravital microscopy has become an important tool to better understand biological processes...
January 19, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29442892/facile-synthesis-of-near-infrared-emissive-cds-quantum-dots-for-live-cells-imaging
#20
Yanyan Cui, Chunfang Zhang, Li Song, Jian Yang, Zhongbo Hu, Xiangfeng Liu
CdS quantum dots (QDs) have attracted extensive attention owing to their great potential in optoelectronic nanodevices and biosensors. But their poor water solubility and high cytotoxicity restrict their practical application in live cell imaging. In addition, CdS QDs usually emit blue or green fluorescence, which also have some limitations for cell imaging due to the "water window" effect. In this study, we report a novel strategy to directly synthesize water-soluble and low-cytotoxic CdS QDs with near infrared (NIR) fluorescence through confinement into BSA templates with the mediation of L-cysteine (Cys)...
April 1, 2018: Journal of Nanoscience and Nanotechnology
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