Analysis of Intracellular Oxygen and Metabolic Responses of Mammalian Cells by Time-Resolved Fluorometry

2007 ◽  
Vol 79 (24) ◽  
pp. 9414-9419 ◽  
Author(s):  
Tomás C. O'Riordan ◽  
Alexander V. Zhdanov ◽  
Gelii V. Ponomarev ◽  
Dmitri B. Papkovsky
Keyword(s):  
Mammalian Cells ◽  
Metabolic Responses ◽  
Time Resolved

Imaging of Cellular Oxygen and Analysis of Metabolic Responses of Mammalian Cells

2009 ◽  
pp. 257-273 ◽  
Author(s):  
Andreas Fercher ◽  
Tomas C. O’Riordan ◽  
Alexander V. Zhdanov ◽  
Ruslan I. Dmitriev ◽  
Dmitri B. Papkovsky
Keyword(s):  
Mammalian Cells ◽  
Metabolic Responses ◽  
Cellular Oxygen

scholarly journals Fluorescence-Based Cell Viability Screening Assays Using Water-Soluble Oxygen Probes

2003 ◽  
Vol 8 (3) ◽  
pp. 264-272 ◽  
Author(s):  
James Hynes ◽  
Suzanne Floyd ◽  
Aleksi E. Soini ◽  
Rosemary O'Connor ◽  
Dmitri B. Papkovsky
Keyword(s):  
Cell Viability ◽  
Mammalian Cells ◽  
Cost Effective ◽  
Water Soluble ◽  
High Throughput Analysis ◽  
Time Resolved ◽  
Low Concentrations ◽  
Cell Line Cell ◽  
Plate Analysis ◽  
Soluble Oxygen

A simple luminescence-based assay for screening the viability of mammalian cells is described, based on the monitoring of cell respiration by means of a phosphorescent water-soluble oxygen probe that responds to changes in the concentration of dissolved oxygen by changing its emission intensity and lifetime. The probe was added at low concentrations (0.3 μM to 0.5 nM) to each sample containing a culture of cells in the wells of a standard 96-well plate. Analysis of oxygen consumption was initiated by applying a layer of mineral oil on top of each sample followed by monitoring of the phosphorescent signal on a prompt or time-resolved fluorescence plate reader. Rates of oxygen uptake could be determined on the basis of kinetic changes of the phosphorescence (initial slopes) and correlated with cell numbers (105 to 107 cells/mL for FL5.12 lymphoblastic cell line), cell viability, or drug/effector action using appropriate control samples. The assay is cell noninvasive, more simple, robust, and cost-effective than existing microplate-based cell viability assays; is compatible with existing instrumentation; and allows for high-throughput analysis of cell viability. ( Journal of Biomolecular Screening 2003:264-272)

Molecular Mobility in Trehalose Loaded Mammalian Cells: Time-Resolved Fluorescence Anisotropy Measurements

2008 ◽  
Author(s):  
Nilay Chakraborty ◽  
Wesley Parker ◽  
Kevin E. Elliott ◽  
Stuart T. Smith ◽  
Patrick J. Moyer ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Fluid Phase ◽  
Water Soluble ◽  
Great Promise ◽  
Time Resolved ◽  
Intracellular Space ◽  
Membrane Pores ◽  
Fluid Phase Endocytosis ◽  
Membrane Bound ◽  
Cellular Compartments

Many preservation methods have utilized sugars such as trehalose as protectants against injury during cell preservation processing, especially during drying (1–5). As mammalian cells do not synthesize trehalose, research in the mammalian cell desiccation field has focused on the development of strategies to enable trehalose delivery into the intracellular milieu. Numerous techniques have been explored ranging from microinjection (2) to the creation or utilization of membrane pores (1,3). Fluid phase endocytosis has shown great promise as an effective strategy for non-invasively delivering water-soluble materials into the intracellular space (4, 5). In this technique trehalose is transported across the cell membrane in membrane-bound cellular compartments called endosomes. Cells incubated in cell culture medium containing trehalose have been shown to take up considerable amounts of trehalose by this technique (4, 5). How much of this trehalose actually become available for protection of biomolecules during the dehydration process has yet to be determined.

Metabolic responses and pathway changes of mammalian cells under different culture conditions with media supplementations

2018 ◽  
Vol 34 (3) ◽  
pp. 793-805 ◽  
Author(s):  
Seo-Young Park ◽  
Thomas M. Reimonn ◽  
Cyrus D. Agarabi ◽  
Kurt A. Brorson ◽  
Seongkyu Yoon
Keyword(s):  
Mammalian Cells ◽  
Culture Conditions ◽  
Metabolic Responses

Time-Resolved Oxygen Effects in Irradiated Bacteria and Mammalian Cells: A Rapid-Mix Study

1975 ◽  
Vol 62 (3) ◽  
pp. 498 ◽  
Author(s):  
M. A. Shenoy ◽  
J. C. Asquith ◽  
G. E. Adams ◽  
B. D. Michael ◽  
M. E. Watts
Keyword(s):  
Mammalian Cells ◽  
Time Resolved ◽  
Oxygen Effects

scholarly journals Space and time-resolved gene expression experiments on cultured mammalian cells by a single-cell electroporation microarray

2008 ◽  
Vol 25 (1) ◽  
pp. 55-67 ◽  
Author(s):  
S. Vassanelli ◽  
L. Bandiera ◽  
M. Borgo ◽  
G. Cellere ◽  
L. Santoni ◽  
...  
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Mammalian Cells ◽  
Time Resolved ◽  
Cell Electroporation ◽  
Space And Time ◽  
Single Cell Electroporation

The singlet-oxygen-sensitized delayed fluorescence in mammalian cells: a time-resolved microscopy approach

2015 ◽  
Vol 14 (4) ◽  
pp. 700-713 ◽  
Author(s):  
Marek Scholz ◽  
Anna-Louisa Biehl ◽  
Roman Dědic ◽  
Jan Hála
Keyword(s):  
Excited State ◽  
Singlet Oxygen ◽  
Mammalian Cells ◽  
Delayed Fluorescence ◽  
Fibroblast Cells ◽  
Proof Of Concept ◽  
Time Resolved ◽  
Excited State Lifetimes ◽  
Kinetics Of

Microsecond kinetics of singlet-oxygen-sensitized delayed fluorescence (SOSDF) have been detected from individual living fibroblast cells as a proof-of-concept. These provide valuable information about excited state lifetimes and their changes during PDT-like treatment.

scholarly journals Misfolded GPI-anchored proteins are escorted through the secretory pathway by ER-derived factors

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Eszter Zavodszky ◽  
Ramanujan S Hegde
Keyword(s):  
Flow Cytometry ◽  
Plasma Membrane ◽  
Cell Surface ◽  
Quantitative Proteomics ◽  
Mammalian Cells ◽  
Secretory Pathway ◽  
Flow Cytometry Analysis ◽  
Time Resolved ◽  
Interaction Partners ◽  
Quantitative Flow

We have used misfolded prion protein (PrP*) as a model to investigate how mammalian cells recognize and degrade misfolded GPI-anchored proteins. While most misfolded membrane proteins are degraded by proteasomes, misfolded GPI-anchored proteins are primarily degraded in lysosomes. Quantitative flow cytometry analysis showed that at least 85% of PrP* molecules transiently access the plasma membrane en route to lysosomes. Unexpectedly, time-resolved quantitative proteomics revealed a remarkably invariant PrP* interactome during its trafficking from the endoplasmic reticulum (ER) to lysosomes. Hence, PrP* arrives at the plasma membrane in complex with ER-derived chaperones and cargo receptors. These interaction partners were critical for rapid endocytosis because a GPI-anchored protein induced to misfold at the cell surface was not recognized effectively for degradation. Thus, resident ER factors have post-ER itineraries that not only shield misfolded GPI-anchored proteins during their trafficking, but also provide a quality control cue at the cell surface for endocytic routing to lysosomes.

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