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Live cell imaging

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https://www.readbyqxmd.com/read/28340410/development-of-fusogenic-glass-surfaces-that-impart-spatiotemporal-control-over-macrophage-fusion-direct-visualization-of-multinucleated-giant-cell-formation
#1
James J Faust, Wayne Christenson, Kyle Doudrick, Robert Ros, Tatiana P Ugarova
Implantation of synthetic material, including vascular grafts, pacemakers, etc. results in the foreign body reaction and the formation of multinucleated giant cells (MGCs) at the exterior surface of the implant. Despite the long-standing premise that fusion of mononucleated macrophages results in the formation of MGCs, to date, no published study has shown fusion in context with living specimens. This is due to the fact that optical-quality glass, which is required for the majority of live imaging techniques, does not promote macrophage fusion...
March 2, 2017: Biomaterials
https://www.readbyqxmd.com/read/28340100/glioblastoma-stem-cell-differentiation-into-endothelial-cells-evidenced-through-live-cell-imaging
#2
Xin Mei, Yin-Sheng Chen, Fu-Rong Chen, Shao-Yan Xi, Zhong-Ping Chen
Background.: Glioblastoma cell-initiated vascularization is an alternative angiogenesis called vasculogenic mimicry. However, current knowledge on the mechanism of de novo vessel formation from glioblastoma stem cells (GSCs) is limited. Methods.: Sixty-four glioblastoma samples from patients and 10 fluorescent glioma xenograft samples were examined by immunofluorescence staining for endothelial marker (CD34 and CD31) and glial cell marker (glial fibrillary acidic protein [GFAP]) expression...
March 8, 2017: Neuro-oncology
https://www.readbyqxmd.com/read/28339924/the-trans-golgi-network-and-the-golgi-stacks-behave-independently-during-regeneration-after-brefeldin-a-treatment-in-tobacco-by-2-cells
#3
Yoko Ito, Kiminori Toyooka, Masaru Fujimoto, Takashi Ueda, Tomohiro Uemura, Akihiko Nakano
The trans-Golgi network (TGN) plays an essential role in intracellular membrane trafficking. In plant cells, recent live-cell imaging studies have revealed the dynamic behavior of the TGN independent from the Golgi apparatus. In order to better understand the relationships between the two organelles, we examined their dynamic responses to the reagent brefeldin A (BFA) and their recovery after BFA removal. Golgi markers responded to BFA similarly over a range of concentrations, whereas the behavior of the TGN was BFA concentration dependent...
February 21, 2017: Plant & Cell Physiology
https://www.readbyqxmd.com/read/28339185/red-emission-b-n-s-co-doped-carbon-dots-for-colorimetric-and-fluorescent-dual-mode-detection-of-fe3-ions-in-complex-biological-fluids-and-living-cells
#4
Yinghua Liu, Wenxiu Duan, Wei Song, Juanjuan Liu, Cuiling Ren, Jiang Wu, Dan Liu, Hongli Chen
Colorimetric and fluorescent dual mode detection methods have gained much attention in recent years, however, it is still desirable to develop new colorimetric and fluorescent dual mode nanosensors with more simple preparation procedure, low cost and excellent biocompatibility. Herein, a colorimetric and fluorescent nanosensor based on B, N, S-co-doped carbon dots (BNS-CDs) were synthesized by one-step hydrothermal treatment of 2, 5-diaminobenzenesulfonic acid and 4-aminophenylboronic acid hydrochloride. Using this nanosensor, a highly sensitive assay of Fe3+ in the range of 0...
March 24, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28335209/dye-doped-fluorescent-silica-nanoparticles-for-live-cell-and-in-vivo-bioimaging
#5
REVIEW
Wen-Han Zhang, Xiao-Xiao Hu, Xiao-Bing Zhang
The need for novel design strategies for fluorescent nanomaterials to improve our understanding of biological activities at the molecular level is increasing rapidly. Dye-doped fluorescent silica nanoparticles (SiNPs) emerge with great potential for developing fluorescence imaging techniques as a novel and ideal platform for the monitoring of living cells and the whole body. Organic dye-containing fluorescent SiNPs exhibit many advantages: they have excellent biocompatibility, are non-toxic, highly hydrophilic, optically transparent, size-tunable and easily modified with various biomolecules...
April 27, 2016: Nanomaterials
https://www.readbyqxmd.com/read/28334699/robust-cell-tracking-in-epithelial-tissues-through-identification-of-maximum-common-subgraphs
#6
Jochen Kursawe, Rémi Bardenet, Jeremiah J Zartman, Ruth E Baker, Alexander G Fletcher
Tracking of cells in live-imaging microscopy videos of epithelial sheets is a powerful tool for investigating fundamental processes in embryonic development. Characterizing cell growth, proliferation, intercalation and apoptosis in epithelia helps us to understand how morphogenetic processes such as tissue invagination and extension are locally regulated and controlled. Accurate cell tracking requires correctly resolving cells entering or leaving the field of view between frames, cell neighbour exchanges, cell removals and cell divisions...
November 2016: Journal of the Royal Society, Interface
https://www.readbyqxmd.com/read/28334356/transient-cell-intrinsic-activity-regulates-the-migration-and-laminar-positioning-of-cortical-projection-neurons
#7
Nicolas Hurni, Marta Kolodziejczak, Ugo Tomasello, Joan Badia, Moritz Jacobshagen, Julien Prados, Alexandre Dayer
Neocortical microcircuits are built during development and require the coordinated assembly of excitatory glutamatergic projection neurons (PNs) into functional networks. Neuronal migration is an essential step in this process. In addition to cell-intrinsic mechanisms, external cues including neurotransmitters regulate cortical neuron migration, suggesting that early activity could influence this process. Here, we aimed to investigate the role of cell-intrinsic activity in migrating PNs in vivo using a designer receptor exclusively activated by a designer drug (DREADD) chemogenetic approach...
March 17, 2017: Cerebral Cortex
https://www.readbyqxmd.com/read/28333166/effect-of-probe-diffusion-on-the-sofi-imaging-accuracy
#8
Wim Vandenberg, Peter Dedecker
Live-cell super-resolution fluorescence imaging is becoming commonplace for exploring biological systems, though sample dynamics can affect the imaging quality. In this work we evaluate the effect of probe diffusion on super-resolution optical fluctuation imaging (SOFI), using a theoretical model and numerical simulations based on the imaging of live cells labelled with photochromic fluorescent proteins. We find that, over a range of physiological conditions, fluorophore diffusion results in a change in the amplitude of the SOFI signal...
March 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28328542/an-excited-state-intramolecular-photon-transfer-fluorescence-probe-for-localizable-live-cell-imaging-of-cysteine
#9
Wei Liu, Wen Chen, Si-Jia Liu, Jian-Hui Jiang
Small molecule probes suitable for selective and specific fluorescence imaging of some important but low-concentration intracellular reactive sulfur species such as cysteine (Cys) pose a challenge in chemical biology. We present a readily available, fast-response fluorescence probe CHCQ-Ac, with 2-(5'-chloro-2-hydroxyl-phenyl)-6-chloro-4(3 H)-quinazolinone (CHCQ) as the fluorophore and acrylate group as the functional moiety, that enables high-selectivity and high-sensitivity for detecting Cys in both solution and biological system...
March 22, 2017: Methods and Applications in Fluorescence
https://www.readbyqxmd.com/read/28328541/molecular-engineering-of-two-photon-fluorescent-probes-for-bioimaging-applications
#10
Hong-Wen Liu, Yongchao Liu, Peng Wang, Xiao-Bing Zhang
During the past two decades, two-photon microscopy (TPM), which utilizes two near-infrared photons as the excitation source, has emerged as a novel, attractive imaging tool for biological research. Compared with one-photon microscopy, TPM offers several advantages, such as lowering background fluorescence in living cells and tissues, reducing photodamage to biosamples, and a photobleaching phenomenon, offering better 3D spatial localization, and increasing penetration depth. Small-molecule-based two-photon fluorescent probes have been well developed for the detection and imaging of various analytes in biological systems...
March 22, 2017: Methods and Applications in Fluorescence
https://www.readbyqxmd.com/read/28328200/fluorescence-resonance-energy-transfer-based-dna-tetrahedron-nano-tweezer-for-highly-reliable-detection-of-tumor-related-mrna-in-living-cells
#11
Lei He, Dan-Qing Lu, Hao Liang, Sitao Xie, Can Luo, Miaomiao Hu, Liujun Xu, Xiaobing Zhang, Weihong Tan
Accurate detection and imaging of tumor-related mRNA in living cells hold great promise for early cancer detection. However, currently, most probes designed to image intracellular mRNA confront intrinsic interferences arising from complex biological matrices and resulting in inevitable false-positive signals. To circumvent this problem, a new class of intracellular DNA nanoprobe, termed DNA tetrahedron nano-tweezer (DTNT), was developed to reliably image tumor-related mRNA in living cells based on the FRET (fluorescence resonance energy transfer) "off" to "on" signal readout mode...
March 22, 2017: ACS Nano
https://www.readbyqxmd.com/read/28327181/soluble-oligomeric-amyloid-%C3%AE-induces-calcium-dyshomeostasis-that-precedes-synapse-loss-in-the-living-mouse-brain
#12
Michal Arbel-Ornath, Eloise Hudry, Josiah R Boivin, Tadafumi Hashimoto, Shuko Takeda, Kishore V Kuchibhotla, Steven Hou, Carli R Lattarulo, Arianna M Belcher, Naomi Shakerdge, Pariss B Trujillo, Alona Muzikansky, Rebecca A Betensky, Bradley T Hyman, Brian J Bacskai
BACKGROUND: Amyloid-β oligomers (oAβ) are thought to mediate neurotoxicity in Alzheimer's disease (AD), and previous studies in AD transgenic mice suggest that calcium dysregulation may contribute to these pathological effects. Even though AD mouse models remain a valuable resource to investigate amyloid neurotoxicity, the concomitant presence of soluble Aβ species, fibrillar Aβ, and fragments of amyloid precursor protein (APP) complicate the interpretation of the phenotypes. METHOD: To explore the specific contribution of soluble oligomeric Aβ (oAβ) to calcium dyshomeostasis and synaptic morphological changes, we acutely exposed the healthy mouse brain, at 3 to 6 months of age, to naturally occurring soluble oligomers and investigated their effect on calcium levels using in vivo multiphoton imaging...
March 21, 2017: Molecular Neurodegeneration
https://www.readbyqxmd.com/read/28325928/a-microplate-reader-based-system-for-visualizing-transcriptional-activity-during-in-vivo-microbial-interactions-in-space-and-time
#13
Rosanna C Hennessy, Peter Stougaard, Stefan Olsson
Here, we report the development of a microplate reader-based system for visualizing gene expression dynamics in living bacterial cells in response to a fungus in space and real-time. A bacterium expressing the red fluorescent protein mCherry fused to the promoter region of a regulator gene nunF indicating activation of an antifungal secondary metabolite gene cluster was used as a reporter system. Time-lapse image recordings of the reporter red signal and a green signal from fluorescent metabolites combined with microbial growth measurements showed that nunF-regulated gene transcription is switched on when the bacterium enters the deceleration growth phase and upon physical encounter with fungal hyphae...
March 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28324646/from-snapshots-to-movies-understanding-early-tooth-development-in-four-dimensions
#14
Rebecca Kim, Jeremy B A Green, Ophir D Klein
The developing tooth offers a model for the study of ectodermal appendage organogenesis. The signaling networks that regulate tooth development have been intensively investigated, but how cell biological responses to signaling pathways regulate dental morphogenesis remains an open question. The increasing use of ex vivo imaging techniques has enabled live tracking of cell behaviors over time in high resolution. While recent studies using these techniques have improved our understanding of tooth morphogenesis, important gaps remain that require additional investigation...
March 21, 2017: Developmental Dynamics: An Official Publication of the American Association of Anatomists
https://www.readbyqxmd.com/read/28324613/analysis-of-protein-kinetics-using-fluorescence-recovery-after-photobleaching-frap
#15
Nickolaos Nikiforos Giakoumakis, Maria Anna Rapsomaniki, Zoi Lygerou
Fluorescence recovery after photobleaching (FRAP) is a cutting-edge live-cell functional imaging technique that enables the exploration of protein dynamics in individual cells and thus permits the elucidation of protein mobility, function, and interactions at a single-cell level. During a typical FRAP experiment, fluorescent molecules in a defined region of interest within the cell are bleached by a short and powerful laser pulse, while the recovery of the fluorescence in the region is monitored over time by time-lapse microscopy...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28324612/quantitative-image-analysis-of-single-molecule-mrna-dynamics-in-living-cells
#16
José Rino, Ana C de Jesus, Maria Carmo-Fonseca
Single mRNA molecules can be imaged in living cells by a method that consists in genetically inserting binding sites for a bacteriophage protein in the gene of interest. The resulting reporter transgene is then integrated in the genome of cells that express the phage protein fused to a fluorescent tag. Upon transcription, binding of the fluorescent protein to its target sequence makes the RNA visible. With this approach it is possible to track, in real time, the life cycle of a precursor mRNA at the site of transcription in the nucleus and transport of mature mRNA to the cytoplasm...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28324608/two-color-total-internal-reflection-fluorescence-microscopy-of-exocytosis-in-endocrine-cells
#17
Adam J Trexler, Justin W Taraska
We describe a comprehensive method for imaging and analysis of local protein dynamics at single sites of exocytosis in living cultured endocrine cells. This method is well suited to quantitatively map the complex dynamics of individual molecules at single sites of vesicle fusion in live cells.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28324604/imaging-the-dynamics-of-cell-wall-polymer-deposition-in-the-unicellular-model-plant-penium-margaritaceum
#18
David Domozych, Anna Lietz, Molly Patten, Emily Singer, Berke Tinaz, Sandra C Raimundo
The unicellular green alga, Penium margaritaceum, represents a novel and valuable model organism for elucidating cell wall dynamics in plants. This organism's cell wall contains several polymers that are highly similar to those found in the primary cell walls of land plants. Penium is easily grown in laboratory culture and is effectively manipulated in various experimental protocols including microplate assays and correlative microscopy. Most importantly, Penium can be live labeled with cell wall-specific antibodies or other probes and returned to culture where specific cell wall developmental events can be monitored...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28324601/two-photon-intravital-microscopy-animal-preparation-protocol-to-study-cellular-dynamics-in-pathogenesis
#19
Erinke van Grinsven, Chloé Prunier, Nienke Vrisekoop, Laila Ritsma
Two-photon intravital microscopy (2P-IVM) is an advanced imaging platform that allows the visualization of dynamic processes at subcellular resolution in vivo. Dynamic processes like cell migration, cell proliferation, cell-cell interactions, and cell signaling have an interactive character and occur in complex environments. Hence, it is of pivotal importance to study these processes in living animals, using for example 2P-IVM. 2P-IVM can be performed on a variety of tissues, from the skin of the animal to internal organs, and a variety of methods can be utilized to perform 2P-IVM on these tissues...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28324599/three-dimensional-live-imaging-of-filamentous-fungi-with-light-sheet-based-fluorescence-microscopy-lsfm
#20
Francesco Pampaloni, Laura Knuppertz, Andrea Hamann, Heinz D Osiewacz, Ernst H K Stelzer
We describe a method for the three-dimensional live imaging of filamentous fungi with light sheet-based fluorescence microscopy (LSFM). LSFM provides completely new opportunities to investigate the biology of fungal cells and other microorganisms with high spatial and temporal resolution. As an example, we study the established aging model Podospora anserina. The protocol explains the mounting of the live fungi for the light sheet imaging, the imaging procedure and illustrates basic image processing of data...
2017: Methods in Molecular Biology
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