The Experts below are selected from a list of 318 Experts worldwide ranked by ideXlab platform
Gérard Lizard - One of the best experts on this subject based on the ideXlab platform.
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Chronology of cellular alterations during 7-ketocholesterol-induced cell death on A7R5 rat smooth muscle cells: Analysis by time lapse-Video Microscopy and conventional fluorescence Microscopy
Cytometry Part A, 2003Co-Authors: Jean-marie Zahm, Sonia Baconnais, Serge Monier, Noël Bonnet, Ginette Bessède, Philippe Gambert, Edith Puchelle, Gérard LizardAbstract:BACKGROUND: Time-Lapse Video Microscopy was used to determine whether mitochondrial and nuclear changes (decrease in mitochondrial transmembrane potential, condensation, and/or fragmentation of the nuclei, morphologic features typical of apoptosis) occurring during 7-ketocholesterol-induced cell death on A7R5 rat smooth muscle cells took place before or after the loss of cell adhesion. In addition, changes in actin organization were followed by conventional fluorescence Microscopy. METHODS: Morphologic, functional, and spatial changes at the mitochondrial level were investigated with 3,3'-dihexyloxacarbocyanine iodide and/or MitoTracker Red, and nuclear morphology was characterized by staining with Hoechst 33342. Actin fibers, which are major components of the filament network of the cytoskeleton, were visualized with phalloidin linked to fluorescein. The numbers of adherent and nonadherent cells were determined by cell counting. RESULTS: 7-Ketocholesterol-induced cell death was associated with a rapid alteration of actin fibers, a loss of intercellular junctions, and cell shape modifications. Analysis of mitochondrial transmembrane potential showed successively a hyperpolarization and a more or less pronounced progressive decrease followed by a dramatic drop associated with an increase in Hoechst 33342 staining, reflecting chromatin condensation and morphologic changes in the nuclei. CONCLUSIONS: During cell death induced by 7-ketocholesterol in A7R5 rat smooth muscle cells, the different methods of Microscopy allowed us to establish that alterations of actin fibers and mitochondrial dysfunctions occurred before condensation and/or fragmentation of the nuclei, which preceded the loss of cell adhesion
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Chronology of cellular alterations during 7‐ketocholesterol–induced cell death on A7R5 rat smooth muscle cells: Analysis by time lapse‐Video Microscopy and conventional fluorescence Microscopy
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 2003Co-Authors: Jean-marie Zahm, Sonia Baconnais, Serge Monier, Noël Bonnet, Ginette Bessède, Philippe Gambert, Edith Puchelle, Gérard LizardAbstract:Background Time-Lapse Video Microscopy was used to determine whether mitochondrial and nuclear changes (decrease in mitochondrial transmembrane potential, condensation, and/or fragmentation of the nuclei, morphologic features typical of apoptosis) occurring during 7-ketocholesterol–induced cell death on A7R5 rat smooth muscle cells took place before or after the loss of cell adhesion. In addition, changes in actin organization were followed by conventional fluorescence Microscopy. Methods Morphologic, functional, and spatial changes at the mitochondrial level were investigated with 3,3′-dihexyloxacarbocyanine iodide and/or MitoTracker Red, and nuclear morphology was characterized by staining with Hoechst 33342. Actin fibers, which are major components of the filament network of the cytoskeleton, were visualized with phalloidin linked to fluorescein. The numbers of adherent and nonadherent cells were determined by cell counting. Results 7-Ketocholesterol–induced cell death was associated with a rapid alteration of actin fibers, a loss of intercellular junctions, and cell shape modifications. Analysis of mitochondrial transmembrane potential showed successively a hyperpolarization and a more or less pronounced progressive decrease followed by a dramatic drop associated with an increase in Hoechst 33342 staining, reflecting chromatin condensation and morphologic changes in the nuclei. Conclusions During cell death induced by 7-ketocholesterol in A7R5 rat smooth muscle cells, the different methods of Microscopy allowed us to establish that alterations of actin fibers and mitochondrial dysfunctions occurred before condensation and/or fragmentation of the nuclei, which preceded the loss of cell adhesion. Cytometry Part A 52A:57–69, 2003. © 2003 Wiley-Liss, Inc.
Peter Neuhaus - One of the best experts on this subject based on the ideXlab platform.
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Visualization of liver sinusoidal endothelial cell repair behavior after preservation by in vitro Time-Lapse Video Microscopy.
Transplantation, 1997Co-Authors: Jörg C. Gerlach, Ralf A. Ziemer, Gabriele Spatkowski, Katrin Zeilinger, Peter NeuhausAbstract:Sinusoidal endothelial cells are significantly more vulnerable to cold storage and reperfusion than hepatocytes. In this study, a method for assessing the repair behavior of sinusoidal endothelial cells in vitro, after preservation, was investigated. Time-Lapse Video Microscopy analysis was performed and migration rates, division rates, and cell detachment rates were determined. Preservation intervals between 3 and 24 hr and reoxygenation times between 4 and 24 hr were compared. A comparison between sinusoidal endothelial cultures that were stored for 6 hr in University of Wisconsin solution and nonpreserved control cultures was performed. This method allows the investigation of the repair capability of individual cells in vitro. Indications of the kind of preservation/reoxygenation injury that occurs after treatment with several preservation solutions and the resultant repair behavior can be obtained.
Jean-marie Zahm - One of the best experts on this subject based on the ideXlab platform.
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Chronology of cellular alterations during 7-ketocholesterol-induced cell death on A7R5 rat smooth muscle cells: Analysis by time lapse-Video Microscopy and conventional fluorescence Microscopy
Cytometry Part A, 2003Co-Authors: Jean-marie Zahm, Sonia Baconnais, Serge Monier, Noël Bonnet, Ginette Bessède, Philippe Gambert, Edith Puchelle, Gérard LizardAbstract:BACKGROUND: Time-Lapse Video Microscopy was used to determine whether mitochondrial and nuclear changes (decrease in mitochondrial transmembrane potential, condensation, and/or fragmentation of the nuclei, morphologic features typical of apoptosis) occurring during 7-ketocholesterol-induced cell death on A7R5 rat smooth muscle cells took place before or after the loss of cell adhesion. In addition, changes in actin organization were followed by conventional fluorescence Microscopy. METHODS: Morphologic, functional, and spatial changes at the mitochondrial level were investigated with 3,3'-dihexyloxacarbocyanine iodide and/or MitoTracker Red, and nuclear morphology was characterized by staining with Hoechst 33342. Actin fibers, which are major components of the filament network of the cytoskeleton, were visualized with phalloidin linked to fluorescein. The numbers of adherent and nonadherent cells were determined by cell counting. RESULTS: 7-Ketocholesterol-induced cell death was associated with a rapid alteration of actin fibers, a loss of intercellular junctions, and cell shape modifications. Analysis of mitochondrial transmembrane potential showed successively a hyperpolarization and a more or less pronounced progressive decrease followed by a dramatic drop associated with an increase in Hoechst 33342 staining, reflecting chromatin condensation and morphologic changes in the nuclei. CONCLUSIONS: During cell death induced by 7-ketocholesterol in A7R5 rat smooth muscle cells, the different methods of Microscopy allowed us to establish that alterations of actin fibers and mitochondrial dysfunctions occurred before condensation and/or fragmentation of the nuclei, which preceded the loss of cell adhesion
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Chronology of cellular alterations during 7‐ketocholesterol–induced cell death on A7R5 rat smooth muscle cells: Analysis by time lapse‐Video Microscopy and conventional fluorescence Microscopy
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 2003Co-Authors: Jean-marie Zahm, Sonia Baconnais, Serge Monier, Noël Bonnet, Ginette Bessède, Philippe Gambert, Edith Puchelle, Gérard LizardAbstract:Background Time-Lapse Video Microscopy was used to determine whether mitochondrial and nuclear changes (decrease in mitochondrial transmembrane potential, condensation, and/or fragmentation of the nuclei, morphologic features typical of apoptosis) occurring during 7-ketocholesterol–induced cell death on A7R5 rat smooth muscle cells took place before or after the loss of cell adhesion. In addition, changes in actin organization were followed by conventional fluorescence Microscopy. Methods Morphologic, functional, and spatial changes at the mitochondrial level were investigated with 3,3′-dihexyloxacarbocyanine iodide and/or MitoTracker Red, and nuclear morphology was characterized by staining with Hoechst 33342. Actin fibers, which are major components of the filament network of the cytoskeleton, were visualized with phalloidin linked to fluorescein. The numbers of adherent and nonadherent cells were determined by cell counting. Results 7-Ketocholesterol–induced cell death was associated with a rapid alteration of actin fibers, a loss of intercellular junctions, and cell shape modifications. Analysis of mitochondrial transmembrane potential showed successively a hyperpolarization and a more or less pronounced progressive decrease followed by a dramatic drop associated with an increase in Hoechst 33342 staining, reflecting chromatin condensation and morphologic changes in the nuclei. Conclusions During cell death induced by 7-ketocholesterol in A7R5 rat smooth muscle cells, the different methods of Microscopy allowed us to establish that alterations of actin fibers and mitochondrial dysfunctions occurred before condensation and/or fragmentation of the nuclei, which preceded the loss of cell adhesion. Cytometry Part A 52A:57–69, 2003. © 2003 Wiley-Liss, Inc.
Richard Gordon - One of the best experts on this subject based on the ideXlab platform.
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Surface contraction and expansion waves correlated with differentiation in axolotl embryos. II. In contrast to urodeles, the anuran Xenopus laevis does not show furrowing surface contraction waves.
The International journal of developmental biology, 1996Co-Authors: P D Nieuwkoop, Natalie K. Björklund, Richard GordonAbstract:We have observed a number of contraction waves traversing the axolotl (Ambystoma mexicanum) embryo (a urodelan amphibian) from the midblastula transition up to at least neural tube closure, and wished to learn if similar "differentiation waves" appear on the popular laboratory anuran amphibian, the South African clawed toad, Xenopus laevis. Time lapse Video Microscopy showed that no contraction waves are visible on the surface of Xenopus from gastrulation through neurulation. It is possible that cell intercalations in the double-layered ectoderm of the Xenopus embryo are homologous to the surface waves in the single layered ectoderm of the axolotl embryo. In any case, a simple, universal correspondence between surface waves and induction phenomena and differentiation does not exist.
Felipe Ortega - One of the best experts on this subject based on the ideXlab platform.
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Time-Lapse Video Microscopy and Single Cell Tracking to Study Neural Cell Behavior In Vitro.
Methods in molecular biology (Clifton N.J.), 2019Co-Authors: Lucía Paniagua-herranz, Rosa Gómez-villafuertes, David De Agustín-durán, Sergio Gascón, Raquel Pérez-sen, Esmerilda G. Delicado, María Teresa Miras-portugal, Felipe OrtegaAbstract:A comprehensive understanding of the mechanisms controlling the behavior of cell populations with regenerative potential is the first step to design effective therapeutic strategies for many diseases. However, a precise description of the biological events involved, such as proliferation, differentiation, cell fate decisions, migration, or viability, may be hampered by the classical use of experiments based on end-point analysis. By contrast, live imaging and single cell tracking provides researchers with an accurate readout of these features in cells throughout an experiment. Here, we describe a protocol to apply Time-Lapse Video Microscopy and post-processing of the data to study critical aspects of the biology and the lineage progression of multiple neural populations.
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Commentary: Live Imaging Followed by Single Cell Tracking to Monitor the Cell Biology and Lineage Progression of Multiple Neural Populations
Journal of Neurology & Neuromedicine, 2018Co-Authors: Felipe Ortega, Madrid, SpainAbstract:Live imaging and single cell tracking enables researchers to monitor crucial aspects of the biology of neural populations. In this commentary, we highlight the requirements, applications, and limitations of a protocol recently published by our research group. This protocol involves adapting the culture of several types of neural cells to Time-Lapse Video Microscopy, and the post-processing of the data to track distinct cell populations.
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Using an adherent cell culture of the mouse subependymal zone to study the behavior of adult neural stem cells on a single-cell level
Nature Protocols, 2011Co-Authors: Felipe Ortega, Timm Schroeder, Marcos R. Costa, Tatiana Simon-ebert, Magdalena Götz, Benedikt BerningerAbstract:A comprehensive understanding of the cell biology of adult neural stem cells (aNSCs) requires direct observation of aNSC division and lineage progression in the absence of niche-dependent signals. Here we describe a culture preparation of the adult mouse subependymal zone (SEZ), which allows for continuous single-cell tracking of aNSC behavior. The protocol involves the isolation (∼3 h) and culture of cells from the adult SEZ at low density in the absence of mitogenic growth factors in chemically defined medium and subsequent live imaging using Time-Lapse Video Microscopy (5–7 d); these steps are followed by postimaging immunocytochemistry to identify progeny (∼7 h). This protocol enables the observation of the progression from slow-dividing aNSCs of radial/astroglial identity up to the neuroblast stage, involving asymmetric and symmetric cell divisions of distinct fast-dividing precursors. This culture provides an experimental system for studying instructive or permissive effects of signal molecules on aNSC modes of cell division and lineage progression.