Document Detail


Concomitant real-time monitoring of intracellular reactive oxygen species and mitochondrial membrane potential in individual living promonocytic cells.
MedLine Citation:
PMID:  17011571     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Reactive oxygen species (ROS) have recently been shown to be involved in multiple physiological responses through modulation of signaling pathways. Inappropriate production of these radicals, and their metabolites, leads to the development of various pathologies. Free radicals can induce both positive and negative effects in cells, and their metabolic pathways are very complex. Hence, it is crucial to be able to simultaneously and directly determine their production dynamics and concentrations in individual living cells, in physiological or pathological states, and in response to drugs. The aim of the present study was to monitor in real time the rates of ROS generation in promonocytic cells upon stimulation with hydrogen peroxide and oxidized lipid. Quantitative detection of intracellular ROS concentration in intact living U937 cells was performed by fluorescence intensity (FI) and polarization (FP) measurements utilizing the Optical LiveCell Array technology. The "dihydro" derivative probes of fluorescein (DCF-DA) and rhodamine (DHR123) were used to assess the intracellular levels of ROS. Each probe molecule exhibited a characteristic FI and FP in its non-fluorescent or oxidized form. Analysis of the temporal relationship between the kinetics of ROS generation and the onset of changes in mitochondrial membrane potential shows high variability within a cell population with regard to both processes. The data demonstrated that temporal measurement of ROS generation, in identifiable individual cells, reveals kinetic behavior that otherwise would be undetected.
Authors:
Naomi Zurgil; Yana Shafran; Elena Afrimzon; Dror Fixler; Asher Shainberg; Mordechai Deutsch
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-08-23
Journal Detail:
Title:  Journal of immunological methods     Volume:  316     ISSN:  0022-1759     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  2006 Oct 
Date Detail:
Created Date:  2006-10-23     Completed Date:  2006-12-19     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  27-41     Citation Subset:  IM    
Affiliation:
The Biophysical Interdisciplinary Jerome Schottenstein Center for the Research and the Technology of the Cellome, Department of Physics, Bar-Ilan University, Ramat Gan 52900, Israel.
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MeSH Terms
Descriptor/Qualifier:
Dicarbethoxydihydrocollidine / analogs & derivatives,  blood
Fluoresceins / chemistry
Humans
Hydrogen Peroxide / pharmacology
Lipopolysaccharides / pharmacology
Membrane Potentials
Microscopy, Fluorescence
Microscopy, Polarization
Mitochondrial Membranes / metabolism,  physiology*
Reactive Oxygen Species / analysis*,  metabolism
Rhodamine 123 / chemistry
Tissue Array Analysis / methods*
U937 Cells
Chemical
Reg. No./Substance:
0/Fluoresceins; 0/Lipopolysaccharides; 0/Reactive Oxygen Species; 0/dihydroethidine; 596-09-8/3',6'-diacetylfluorescein; 62669-70-9/Rhodamine 123; 632-93-9/Dicarbethoxydihydrocollidine; 7722-84-1/Hydrogen Peroxide

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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