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

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

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.

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.

Ultrastructural Alterations in Hela Cells Induced by Spider Venom

1967 ◽  
Vol 25 ◽  
pp. 20-21
Author(s):  
Dale E. McClendon ◽  
Paul N. Morgan ◽  
Bernard L. Soloff
Keyword(s):  
Growth Medium ◽  
Mammalian Cells ◽  
Carcinoma Cells ◽  
Venom Protein ◽  
Sterile Saline ◽  
Carbon Coated ◽  
Available Information ◽  
Loxosceles Reclusa ◽  
Essential Growth ◽  
Solution Cultures

It has been observed that minute amounts of venom from the brown recluse spider, Loxosceles reclusa, are capable of producing cytotoxic changes in cultures of certain mammalian cells (Morgan and Felton, 1965). Since there is little available information concerning the effect of venoms on susceptible cells, we have attempted to characterize, at the electron microscope level, the cytotoxic changes produced by the venom of this spider.Cultures of human epithelial carcinoma cells, strain HeLa, were initiated on sterile, carbon coated coverslips contained in Leighton tubes. Each culture was seeded with approximately 1x105 cells contained in 1.5 ml of a modified Eagle's minimum essential growth medium prepared in Hank's balanced salt solution. Cultures were incubated at 36° C. for three days prior to the addition of venom. The venom was collected from female brown recluse spiders and diluted in sterile saline. Protein determinations on the venom-were made according to the spectrophotometric method of Waddell (1956). Approximately 10 μg venom protein per ml of fresh medium was added to each culture after discarding the old growth medium. Control cultures were treated similarly, except that no venom was added. All cultures were reincubated at 36° C.

Effects of Hyperoxia on Lung Utrastructure

1970 ◽  
Vol 28 ◽  
pp. 26-27
Author(s):  
Gladys Harrison
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Light Microscopy ◽  
Oxygen Effects ◽  
Age Dependent ◽  
The Central Nervous System ◽  
The Subject ◽  
Space Age ◽  
Alveolar Septa ◽  
Extensive Damage

With the advent of the space age and the need to determine the requirements for a space cabin atmosphere, oxygen effects came into increased importance, even though these effects have been the subject of continuous research for many years. In fact, Priestly initiated oxygen research when in 1775 he published his results of isolating oxygen and described the effects of breathing it on himself and two mice, the only creatures to have had the “privilege” of breathing this “pure air”.Early studies had demonstrated the central nervous system effects at pressures above one atmosphere. Light microscopy revealed extensive damage to the lungs at one atmosphere. These changes which included perivascular and peribronchial edema, focal hemorrhage, rupture of the alveolar septa, and widespread edema, resulted in death of the animal in less than one week. The severity of the symptoms differed between species and was age dependent, with young animals being more resistant.

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