Fluorescence enhancement of DNA-bound TO-PRO-3 by incorporation of bromodeoxyuridine to monitor cell cycle kinetics

Beisker, Wolfgang; Weller‐Mewe, Eva Maria; Nüsse, Michael

doi:10.1002/(sici)1097-0320(19991101)37:3<221::aid-cyto9>3.0.co;2-1

Cited by 19 publications

(9 citation statements)

References 24 publications

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“…In general, signal intensity for TT3 appeared slightly lower compared with TP3, and addition of SDS tothe staining buffer resulted in slightly reduced fluorescence. On the other hand, TP3 showed higher sensitivity to photo- bleaching, which agrees with published results for eukaryotic cells (13,14). …”

supporting

confidence: 92%

The Fluorescent Dyes TO-PRO-3 and TOTO-3 Iodide Allow Detection of Microbial Cells in Soil Samples without Interference From Background Fluorescence

Henneberger

Birch²,

Bergquist³

et al. 2011

BioTechniques

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Visualization of microorganisms in soils and sediments using fluorescent dyes is a common method in microbial ecology studies, but is often hampered by strong nonspecific background fluorescence that can mask genuine cellular signals. The cyanine nucleic acid binding dyes TO-PRO-3 and TOTO-3 iodide enabled a clear detection of microbial cells in a mineral soil, while nonspecific background was greatly reduced compared with commonly used dyes. When used as counterstains for fluorescence in situ hybridization (FISH), both cyanine dyes allowed identification of microbial cells despite strong background from nonspecifically bound probes. TO-PRO-3 and TOTO-3 are easy to use and represent superior alternatives for detecting microorganisms in soil environments.

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supporting

confidence: 92%

The Fluorescent Dyes TO-PRO-3 and TOTO-3 Iodide Allow Detection of Microbial Cells in Soil Samples without Interference From Background Fluorescence

Henneberger

Birch²,

Bergquist³

et al. 2011

BioTechniques

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“…27 If DNA is labeled by both PI and TO-PRO-3, however, fluorescence resonance energy transfer (FRET) occurs from PI to TO-PRO-3 with fluorescence excitation at 530 nm. 29,30 This interaction decreases the fluorescence intensity of PI and increases the fluorescence intensity of TO-PRO-3 (Online Figure VI). …”

Section: Resultsmentioning

confidence: 94%

Tracking Chromatid Segregation to Identify Human Cardiac Stem Cells That Regenerate Extensively the Infarcted Myocardium

Kajstura

Bai

Cappetta

et al. 2012

Circulation Research

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Rationale According to the immortal DNA strand hypothesis, dividing stem cells selectively segregate chromosomes carrying the old template DNA, opposing accumulation of mutations resulting from non-repaired replication errors and attenuating telomere shortening. Objective Based on the premise of the immortal DNA strand hypothesis, we propose that stem cells retaining the old DNA would represent the most powerful cells for myocardial regeneration. Methods and Results Division of hCSCs by non-random and random segregation of chromatids was documented by clonal assay of bromodeoxyuridine-tagged hCSCs. Additionally, their growth properties were determined by a series of in vitro and in vivo studies. We report that a small class of hCSCs retain during replication the mother DNA and generate two daughter cells, which carry the old and new DNA, respectively. hCSCs with immortal DNA form a pool of non-senescent cells with longer telomeres and higher proliferative capacity. The self-renewal and long-term repopulating ability of these cells was shown in serial-transplantation assays in the infarcted heart; these cells created a chimeric organ, composed of spared rat and regenerated human cardiomyocytes and coronary vessels, leading to a remarkable restoration of cardiac structure and function. The documentation that hCSCs divide by asymmetric and symmetric chromatid segregation supports the view that the human heart is a self-renewing organ regulated by a compartment of resident hCSCs. Conclusions The impressive recovery in ventricular hemodynamics and anatomy mediated by clonal hCSCs carrying the “mother” DNA underscores the clinical relevance of this stem cell class for the management of heart failure in humans.

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“…Cell Proliferation assays were performed as described (Beisker et al, 1999). Briefly, 10 mM bromodeoxyuridine (BrdU) was added to cerebellar slices each day.…”

Section: Cell Proliferation Assaymentioning

confidence: 99%

TGF-β2 neutralization inhibits proliferation and activates apoptosis of cerebellar granule cell precurors in the developing cerebellum

Elvers¹,

Pfeiffer²,

Kaltschmidt³

et al. 2005

Mechanisms of Development

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Transforming growth factor beta 2 (TGF-beta2) plays a critical role in growth, differentiation and cell death, but its function in the developing cerebellum is still uncertain. In this study we analyzed the effects of TGF-beta2 on ex vivo developing cerebellar slice cultures. Proliferation of granule cell precursors peaked ex vivo in the same developmental window as in vivo (P8-P14). Addition of recombinant TGF-beta2 could extent the proliferation of granule cell precursors and induced a second late proliferation wave. In contrast, antibody neutralization of TGF-beta2 strongly reduced proliferation and induced neurodegeneration. TGF-beta2 neutralization resulted in apoptotic cells, which showed caspase 3 activation. Taken together our results demonstrate that TGF-beta2 is a novel growth and survival factor for granule cells precursors in the developing cerebellum.

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Fluorescence enhancement of DNA-bound TO-PRO-3 by incorporation of bromodeoxyuridine to monitor cell cycle kinetics

Cited by 19 publications

References 24 publications

The Fluorescent Dyes TO-PRO-3 and TOTO-3 Iodide Allow Detection of Microbial Cells in Soil Samples without Interference From Background Fluorescence

The Fluorescent Dyes TO-PRO-3 and TOTO-3 Iodide Allow Detection of Microbial Cells in Soil Samples without Interference From Background Fluorescence

Tracking Chromatid Segregation to Identify Human Cardiac Stem Cells That Regenerate Extensively the Infarcted Myocardium

TGF-β2 neutralization inhibits proliferation and activates apoptosis of cerebellar granule cell precurors in the developing cerebellum

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