scholarly journals Fluorescence Live-cell Imaging of the Complete Vegetative Cell Cycle of the Slow-growing Social Bacterium Myxococcus xanthus

10.3791/57860 ◽  
2018 ◽  
Author(s):  
Dominik Schumacher ◽  
Lotte Søgaard-Andersen
Keyword(s):  
Cell Cycle ◽  
Vegetative Cell ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Myxococcus Xanthus ◽  
Live Cell ◽  
Slow Growing ◽  
Fluorescence Live Cell Imaging

scholarly journals Live-cell imaging to detect phosphatidylserine externalization in brain endothelial cells exposed to ionizing radiation: implications for the treatment of brain arteriovenous malformations

2016 ◽  
Vol 124 (6) ◽  
pp. 1780-1787 ◽  
Author(s):  
Zhenjun Zhao ◽  
Michael S. Johnson ◽  
Biyi Chen ◽  
Michael Grace ◽  
Jaysree Ukath ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Endothelial Cells ◽  
Ionizing Radiation ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Vascular Targeting ◽  
Radiation Doses ◽  
Radiation Induced ◽  
Polarity Sensitive

OBJECT Stereotactic radiosurgery (SRS) is an established intervention for brain arteriovenous malformations (AVMs). The processes of AVM vessel occlusion after SRS are poorly understood. To improve SRS efficacy, it is important to understand the cellular response of blood vessels to radiation. The molecular changes on the surface of AVM endothelial cells after irradiation may also be used for vascular targeting. This study investigates radiation-induced externalization of phosphatidylserine (PS) on endothelial cells using live-cell imaging. METHODS An immortalized cell line generated from mouse brain endothelium, bEnd.3 cells, was cultured and irradiated at different radiation doses using a linear accelerator. PS externalization in the cells was subsequently visualized using polarity-sensitive indicator of viability and apoptosis (pSIVA)-IANBD, a polarity-sensitive probe. Live-cell imaging was used to monitor PS externalization in real time. The effects of radiation on the cell cycle of bEnd.3 cells were also examined by flow cytometry. RESULTS Ionizing radiation effects are dose dependent. Reduction in the cell proliferation rate was observed after exposure to 5 Gy radiation, whereas higher radiation doses (15 Gy and 25 Gy) totally inhibited proliferation. In comparison with cells treated with sham radiation, the irradiated cells showed distinct pseudopodial elongation with little or no spreading of the cell body. The percentages of pSIVA-positive cells were significantly higher (p = 0.04) 24 hours after treatment in the cultures that received 25- and 15-Gy doses of radiation. This effect was sustained until the end of the experiment (3 days). Radiation at 5 Gy did not induce significant PS externalization compared with the sham-radiation controls at any time points (p > 0.15). Flow cytometric analysis data indicate that irradiation induced growth arrest of bEnd.3 cells, with cells accumulating in the G2 phase of the cell cycle. CONCLUSIONS Ionizing radiation causes remarkable cellular changes in endothelial cells. Significant PS externalization is induced by radiation at doses of 15 Gy or higher, concomitant with a block in the cell cycle. Radiation-induced markers/targets may have high discriminating power to be harnessed in vascular targeting for AVM treatment.

scholarly journals “Probe, Sample, and Instrument (PSI)”: The Hat-Trick for Fluorescence Live Cell Imaging

Chemosensors ◽  
2018 ◽  
Vol 6 (3) ◽  
pp. 40 ◽  
Author(s):  
Ludovic Galas ◽  
Thibault Gallavardin ◽  
Magalie Bénard ◽  
Arnaud Lehner ◽  
Damien Schapman ◽  
...  
Keyword(s):  
Organic Synthesis ◽  
Fluorescent Probes ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Cell Labeling ◽  
Key Points ◽  
Research Infrastructures ◽  
New Strategies ◽  
Fluorescence Live Cell Imaging

Cell Imaging Platforms (CIPs) are research infrastructures offering support to a number of scientific projects including the choice of adapted fluorescent probes for live cell imaging. What to detect in what type of sample and for how long is a major issue with fluorescent probes and, for this, the “hat-trick” “Probe–Sample–Instrument” (PSI) has to be considered. We propose here to deal with key points usually discussed in CIPs including the properties of fluorescent organic probes, the modality of cell labeling, and the best equipment to obtain appropriate spectral, spatial, and temporal resolution. New strategies in organic synthesis and click chemistry for accessing probes with enhanced photophysical characteristics and targeting abilities will also be addressed. Finally, methods for image processing will be described to optimize exploitation of fluorescence signals.

scholarly journals Live Cell Imaging of Telomerase RNA Dynamics Reveals Cell Cycle-Dependent Clustering of Telomerase at Elongating Telomeres

2011 ◽  
Vol 44 (5) ◽  
pp. 819-827 ◽  
Author(s):  
Franck Gallardo ◽  
Nancy Laterreur ◽  
Emilio Cusanelli ◽  
Faissal Ouenzar ◽  
Emmanuelle Querido ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Telomerase Rna ◽  
Rna Dynamics ◽  
Cycle Dependent

scholarly journals Long-term fate of etoposide-induced micronuclei and micronucleated cells in Hela-H2B-GFP cells

2020 ◽  
Vol 94 (10) ◽  
pp. 3553-3561
Author(s):  
Hauke Reimann ◽  
Helga Stopper ◽  
Henning Hintzsche
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Cellular Structures ◽  
Death Rates ◽  
Daughter Cells

Abstract Micronuclei are small nuclear cellular structures containing whole chromosomes or chromosomal fragments. While there is a lot of information available about the origin and formation of micronuclei, less is known about the fate of micronuclei and micronucleated cells. Possible fates include extrusion, degradation, reincorporation and persistence. Live cell imaging was performed to quantitatively analyse the fates of micronuclei and micronucleated cells occurring in vitro. Imaging was conducted for up to 96 h in HeLa-H2B-GFP cells treated with 0.5, 1 and 2 µg/ml etoposide. While a minority of micronuclei was reincorporated into the main nucleus during mitosis, the majority of micronuclei persisted without any alterations. Degradation and extrusion were observed rarely or never. The presence of micronuclei affected the proliferation of the daughter cells and also had an influence on cell death rates. Mitotic errors were found to be clearly increased in micronucleus-containing cells. The results show that micronuclei and micronucleated cells can, although delayed in cell cycle, sustain for multiple divisions.

scholarly journals TADF Dye-Loaded Nanoparticles for Fluorescence Live-Cell Imaging

2020 ◽  
Vol 8 ◽  
Author(s):  
Carina I. C. Crucho ◽  
João Avó ◽  
Ana M. Diniz ◽  
Sandra N. Pinto ◽  
José Barbosa ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Fluorescence Live Cell Imaging

Analysis of the influence of subcellular localization of the HIV Rev protein on Rev-dependent gene expression by multi-fluorescence live-cell imaging

2006 ◽  
Vol 312 (4) ◽  
pp. 443-456 ◽  
Author(s):  
Horst Wolff ◽  
Kamyar Hadian ◽  
Manja Ziegler ◽  
Claudia Weierich ◽  
Susanne Kramer-Hammerle ◽  
...  
Keyword(s):  
Gene Expression ◽  
Subcellular Localization ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Rev Protein ◽  
Fluorescence Live Cell Imaging

Concentration-dependent fluorescence live-cell imaging and tracking of intracellular nanoparticles

2011 ◽  
Vol 22 (23) ◽  
pp. 235101 ◽  
Author(s):  
Ji Hye Seo ◽  
Keunchang Cho ◽  
So Yeong Lee ◽  
Sang-Woo Joo
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Fluorescence Live Cell Imaging
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