Document Detail


Molar quantification by flow cytometry of fatty acid binding to cells using dipyrrometheneboron difluoride derivatives.
MedLine Citation:
PMID:  8742176     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Fatty acid analogs of a dipyrrometheneboron difluoride fluorophore (BDY-FA) have recently been developed. Relative to other fluorescent fatty acids, some of these have the advantages of excitation and emission spectra similar to those of fluorescein and of high quantum yield, which permits their use in conventional argon laser cytometry or microscopy. For the cytofluorimetric quantification of BDY-FA analogs, expressed as molecules bound per cell, we have compared the fluorescence of BDY-dodecanoic acid (BDY-C12) with that of fluorescein. Fluorescent beads with different amounts of bound fluorescein were used to calibrate a flow cytometer in order to correlate the fluorescence intensity with the number of fluorescein molecules per bead. In addition, starting from the basic equation defining the relationship between fluorescence and concentration, we have derived another equation which makes it possible to establish, for a given fluorescence, the relative molar concentration of both fluorochromes and, consequently, to express the fluorescence intensity emitted by the BDY-FA as the equivalent number of BDY-FA molecules. As an example of the potential application of this procedure, the time-course and concentration-dependent binding of BDY-C12 to quiescent and mitogen-activated human lymphocytes and to cultured human T-lymphoma cells have been studied. The method described is of general interest as it can also be applied to the flow cytometric or laser scanning microscopic quantification of other fluorescent dyes.
Authors:
A Macho; Z Mishal; J Uriel
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cytometry     Volume:  23     ISSN:  0196-4763     ISO Abbreviation:  Cytometry     Publication Date:  1996 Feb 
Date Detail:
Created Date:  1996-10-10     Completed Date:  1996-10-10     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8102328     Medline TA:  Cytometry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  166-73     Citation Subset:  IM    
Affiliation:
Laboratoire de Chimie des Proteines, Institut de Recherches Scientifiques sur le Cancer, Villejuif, France.
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MeSH Terms
Descriptor/Qualifier:
Boron Compounds*
Calibration
Cells, Cultured
Fatty Acids / metabolism*
Flow Cytometry*
Fluorescein
Fluoresceins
Fluorescent Dyes
Humans
Kinetics
Lymphocytes / metabolism*
Lymphoma, T-Cell / metabolism*,  pathology
Microscopy, Confocal / methods
Reference Standards
Spectrometry, Fluorescence
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Boron Compounds; 0/Fatty Acids; 0/Fluoresceins; 0/Fluorescent Dyes; 2321-07-5/Fluorescein

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


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